Profiles in Preventive Cardiology Dried leaf extract of Tulsi (Ocimum sanctum Linn) reduces

cardiovascular disease risk factors: Results of a double blinded

randomized controlled trial in healthy volunteers.
Shankar Mondal1, PhD, B.R. Mirdha2, MD, M.M. Padhi3, MD, S.C. Mahapatra1, MD
Departments of 1Physiology and 2Microbiology, All India Institute of Medical Sciences (AIIMS), New Delhi, INDIA
3
Central Council for Research in Ayurveda and Siddha, Department of Ayurveda, Yoga & Naturopathy, Unani and Sidhha (AYUSH),
Janakpuri, New Delhi, India

-Dr. Ancel Benjamin Keys

Abstract Key Words

Ancel Benjamin Keys (January 26, 1904 - November 20, heart attacks, as noted in the newspapers' obituary pages,
Herb Tulsi (Ocimum sanctum Linn.) is traditionally used for ● Cardioprotective
2004) was an American scientist who studied the influence and began to study 283 businessmen from the Twin Cities, various ailments and believed to prevent many diseases if
of diet on health. He closely analyzed the epidemiology of conducting examinations and taking blood samples every ● Cholesterol
taken regularly on an empty stomach. Ethanolic extract of
cardiovascular disease (CVD) and was responsible for two five years. It showed that smoking, high blood pressure and dried leaf of Tulsi [Tulsi extract] have been found to have ● Triglycerides
well-known diets: K-rations formulated as balanced meals elevated cholesterol were commonly seen in men who had immunomodulatory properties in many animal experiments ● Medicinal plant
for combat soldiers in World War II and the Mediterranean suffered from heart attacks. After a decade of work, he and few clinical trials. This study was designed to evaluate the
effects of Tulsi extract on biochemical parameters of healthy
diet which was beneficial for heart health. In particular, he found that saturated fat chiefly determined blood volunteers, having no known history of biochemical or
hypothesized that different kinds of dietary fat had cholesterol levels, a breakthrough research. immunological dysfunctions, through a double blinded ■
Introduction
different effects on health. randomized controlled trial. Biochemical parameters of
This lead to the creation of Seven Countries Study, one of healthy volunteers such as lipid profile, liver and kidney Plant Tulsi (Ocimum sanctum Linn.) belongs to the
Dr. Keys graduated from the University of California at the most rigorous and complicated epidemiological studies function tests were monitored in 24 subjects. One capsule of Lamiacae family which is known for its specific aromatic
300 mg Tulsi extract or placebo (sucrose) was administered on
Berkeley and briefly took a management trainee job at undertaken ever. In 1980, the results of the study were empty stomach once daily for 4 weeks followed by a washout fragrance and medicinal values in traditional medicine
Woolworth's but soon was back in school, earning a published in a book by Harvard Press and revealed that the period of 3 weeks. Then the subjects were crossover to other system. Tulsi considered as the sacred plant in Indian
doctorate in biology from the Scripps Institute in La Jolla, Mediterranean diet, rich in fruits and vegetables, pasta, group and continued for another 4 weeks of intervention. Blood subcontinent. Its medicinal properties have been
California. He had a postdoctoral fellowship in bread and olive oil with meat, fish and dairy products used samples were obtained; 1) before starting of capsule intake 2) mentioned in the ancient medicinal texts of various
after completion of 4 weeks 3) after 3 weeks of washout and 4)
Copenhagen, earned a second doctorate in physiology as condiments, was highly protective against heart disease after completion of 4 weeks in the crossover group.
civilizations of the world.1-3
from Kings College in Cambridge, England, and worked even though the diet derived more than 35 percent of its
briefly at the Mayo Clinic. He joined the University of calories from fat. It was found that some of the subjects who had higher levels of Traditionally, it is believed that regular intake of Tulsi
triglycerides and cholesterols than the normal physiological
Minnesota in 1936 and four years later founded its famous values at beginning significantly reduced their cholesterol leaves on empty stomach keeps the person healthy and
Laboratory of Physiological Hygiene, housed under the Dr. Keys with his wife, a biochemist wrote two best-selling levels. There was also a reducing trend found for triglycerides provides longevity. Scientific evidences that are available
bleachers at the university's stadium. Dr. Keys served there books, "Eat Well and Stay Well" and "How to Eat Well and levels. There were no significant changes in the other on various medicinal aspects of this holy plant i.e.
as a director for 33 years. Stay Well the Mediterranean Way." These and a third book parameters tested. In conclusion it can be said that Tulsi antimicrobial, adaptogenic, anti-diabetic, hepato-
they wrote, "The Benevolent Bean," had the kind of recipes extract have shown cardioprotective properties in the study protective, anti-inflammatory, anti-carcinogenic, radio-
population by significantly reducing higher levels of
He led a scientific excursion to the Andes in 1935 to study that Dr. Keys's studies suggested kept heart disease at bay. cholesterol. protective, immunomodulatory, neuro-protective,
the physiological effects of altitude. It was then the War He is survived by his wife, Margaret; a daughter, Carrie cardioprotective, mosquito repellent etc. to name a few.
Department asked him to develop pocket-size nutritionally D'Andrea of Bloomington; and a son, Dr. Henry Keys, of Most of these evidences are based on in-vitro, experimental
4
adequate food rations for World War II paratroopers. The Albany. Another daughter, Martha McLain, was murdered and a few human studies. It is very well known fact that
result was K ration, named for its developer and distributed in 1991 while on vacation in Jamaica with her family. Tulsi have no side effects and experimental studies shown
to hundreds of thousands of American troops during the that LD50 (Lethal Dose-50) of ethanolic extract is ranging
war. On his hundredth birthday, Dr. Keys was asked whether his
diet had contributed to his long life. He answered, "Very
In 1947, he noticed the increasing numbers of deaths from likely, but no proof." Received: 14-02-12; Revised: 27-02-12; Accepted:05-03-12
Disclosures: This article has not received any funding and has no vested commercial interest
Received:22-02-12; Accepted:06-03-12 Acknowledgements: Indian Council of Medical Research [ICMR], New Delhi for Research Associate Fellowship to S. Mondal. Central
Disclosures: This article has not received any funding and has no vested commercial interest Council for Research in Ayurveda and Siddha [CCRAS], Department of AYUSH, Ministry of Health and Family Welfare, Government of India
Acknowledgements: None for proving Tulsi extract and funds. Dabur Pharmaceuticals [P] Ltd, Ghaziabad, U.P., India for providing placebo and capsuling facility.

176 J. Preventive Cardiology Vol. 1 ■

■ No. 4 May 2012
■ J. Preventive Cardiology ■Vol. 1 ■
No. 4 ■ May 2012 177
 Mondal S, et al Tulsi and CVD

from 4600 mg/kg/bw (body weight) to 6200 mg/kg/bw in were the inclusion criteria for the enrollment of healthy Statistical analysis of results significant (intrasubject p=0.057) changes was seen in
5, 6
experimental mice. volunteers; a) age 18-60 years, either sex, b) not on any Tulsi extract treated groups during pre and post
regular medication and c) devoid of any medication at least Two-way ANOVA (Analysis of Variance) for crossover intervention periods. However, normally the fasting blood
Tulsi extract has been found to be cardioprotective in during last one month. Subjects suffering from different design was used to test the statistical significance of the glucose levels does not remain constant and it varies with
experimental animal studies. Long term oral diseases/disorders and/or having any kind of allergy, results. Independent ‘t-test’ was applied to compare the durations of overnight fasting, type of last meal taken along
administration of fresh Tulsi leaves augments cardiac undergone surgery during last one year, organ transplant baseline of two groups. Wilcoxon singed-rank test was with several other factors. Considering the dynamic
endogenous antioxidants and prevents myocardial necrosis recipients, chronic smokers, pregnant and lactating women applied to analyze triglycerides levels. The results were behavior of blood glucose levels under physiological
models in rats.7-9 Suanarunsawat et al (2011),10 conducted were excluded from the study. Enrolled individuals were considered significant if the p-value was 0.05 for period conditions this 1.56 mg/dl change may be ignored
animal experiments to investigate the lipid-lowering and randomized into two groups. Subjects and the staff directly effects and intervention effects. However, p-value of 0.10 clinically.
11
antioxidative activities of Ocimum sanctum L. (OS) leaf involved with the study subjects and data analysis were for carryover effects was considered significant. All
extracts in liver and heart of rats fed with high-cholesterol blinded about the interventional capsules. Allocation of statistical analysis was carried out using Stata 9.0 software Liver Function Tests (LFT) and Renal Function Tests
(HC) diet for seven weeks. group was concealed in opaque envelope. (Statacorp, Texas, USA). The blinding was decoded only (RFT)
after the statistical analysis.
Their results showed that OS suppressed the high levels of Tulsi extract and placebo administration Liver function tests were monitored by measuring SGOT
serum lipid profile and hepatic lipid content without ■
Results [Serum glutamate oxaloacetate transferase], SGPT [Serum
significant effects on fecal lipid excretion. Fecal bile acids Ethanolic (70%) extract of Ocimum sanctum Linn. leaves; glutamate pyruvate transferase] and also total bilirubin
excretion was increased in HC rats treated with OS. The ‘Tulsi extract’ was supplied by the Central Council for The volunteers of two groups were comparable on basic levels of serum in few subjects. It was found that there were
high serum levels of TBARS [Thiobarbituric Acid Research in Ayurveda and Siddha, Department of AYUSH, anthropometric and physiological parameters. We had a no significant differences during pre and post intervention
Reactive Substances] as well as AST [Aspartate Ministry of Family Health and Welfare, Government of wide range of age in inclusion criteria but subjects in periods in either group. There were no significant changes
Aminotransferase], ALT [Alanine Aminotransferase], AP India. Placebo (sucrose, nutrition grade) was supplied by relatively young age ranging from 22 to 37 years in the blood urea nitrogen (p=0.511) serum protein
[Alkaline Phosphatase], LDH [Lactate Dehydrogenase], the Dabur Pharmaceutical (India) Ltd, Ghaziabad (U.P.), participated in the study. The subjects had a healthy BMI (p=0.838) and creatinine (p=0.766) levels in either the
CK-MB [Creatine Kinase MB subunit] significantly India. Capsulation facilities were provided by the Dabur ranging from 17.3 to 25.64 kg/m2. Clinical biochemistry placebo or the Tulsi extract groups (Table 2).
decreased in HC rats treated with OS. OS treatment also Pharmaceutical to make 300 mg capsules using Tulsi parameters of the subjects were monitored at both pre and
suppressed the high level of TBARS and raised the low extract or placebo. Packaging of both, Tulsi extract and post intervention periods during the study. Total
activities of GPx [Glutathione Peroxides] and CAT placebo, were similar to avoid identification. The cholesterol (TC), triglycerides (TG), fasting blood glucose Triglycerides and total Cholesterol
[Catalase] without any impact on SOD [Superoxide intervention was allocated as per the randomized sequence levels (FBG), liver function test, renal function tests were
Dismutase] in the liver. Histopathological results also once the volunteer met the inclusion criteria and given a recorded. Table 1 shows the comparison of basic and There was no significant (p=0.128 & p=0.232) difference
confirmed that OS preserved the liver and myocardial written informed consent. Total period of trial was of 11 biochemical parameters of placebo and Tulsi extract at the baseline in triglycerides and cholesterol in both the
tissues. Present study was undertaken to validate the weeks in which two active administration period of 4 intervention groups at baseline. groups respectively (Table 2, 3). However it was found that
traditional claims of Tulsi regarding its usefulness in weeks each separated with a three weeks washout period in few individual who otherwise had an apparently normal
general health protection and to monitor various between. One capsule [code named, having Tulsi extract or Independent t-test applied. medical history had high cholesterol and triglycerides
biochemical parameters of healthy volunteers after placebo] of 300 mg was administered once daily, as per levels. It was observed that there was a significant decrease
administration of 300 mg of Tulsi extract or placebo on randomization, on empty stomach for four weeks followed Fasting blood glucose in the elevated levels of cholesterol (intra-subject p=0.003)
empty stomach for a period of 4 weeks. by a wash out period of three weeks before the volunteer of these individuals (n=6) in the Tulsi extract group and this
was crossover for the next intervention. There was a difference of about 1.56 mg/dl decrease in the decrease did not continue when subjects crossed-over to
■
Material and methods Tulsi extract treated groups (Table 2). When two-way the placebo group.
Collection of blood samples ANOVA for crossover design was applied a statistically
Design of the study
Venous blood samples of 5 ml were collected at four
Table 1: Comparison of Anthropometric and Physiological Parameters at
This was a double blind randomized controlled trial of total different time points i.e. i) before administration of
Baseline (before administration of capsules)
11 weeks in a crossover format which included a 3 weeks capsule, ii) after administration of placebo or Tulsi extract
wash-out period, to study the effects of ethanolic extract of capsules for four weeks iii) after the end of three weeks Basic parameters Baseline of Placebo-Tulsi Baseline of Tulsi- Placebo P-Value
Tulsi leaf on clinical biochemistry parameters of healthy washout period and finally iv) after completion of four sequence (n=12) sequence (n=10)
volunteers. The study was approved by the institutional weeks in crossover intervention period. The sampling was Age (years) 27.5 ±
4.75 (22-37) 26.5 ±
3.43 (23-35) 0.585
ethics committee on research involving human subjects done in fasting state [at least 12h] between 8:30 am to 10:00
and the trial was registered with Clinical Trial Registry of am at Department of Physiology, AIIMS, New Delhi. Height (cm) 166.16 ±
7.62 (153-180) 166.1 ±
5.40 (157-176) 0.981
India (No. CTRI/2009/091/000350). Subjects were contacted personally or telephonically twice Weight (kg) 62.5 ±
10.52 (50-80) 65.8 ±
6.05(54-78) 0.391
a week for better compliance and unused capsules were
2
Screening of Volunteers taken back from the subjects. Two volunteers did not BMI (kg/m ) 22.36 ±
2.52(17.3-25.43) 23.80 ±
1.13 (21.91-25.64) 0.111
continue the intervention due to their personal choice. Systolic BP (mm Hg) 109.16 ±
7.50 (100-120) 113.2 ±
7.37 (100-124) 0.220
Forty five volunteers were screened to enroll in the study. Clinical biochemical parameters were studied using
Twenty four healthy volunteers, who met the inclusion commercially available kits [Transasia, India; Randox, Diastolic BP (mm Hg) 74.16 ±
4.62 (68-80) 76.2 ±
3.45 (70-80) 0.265
criteria and gave their consent, were enrolled. Following United Kingdom] and semi-automatic biochemistry All values presented in mean ±
SD (range).P-value £
0.05 was considered as significant.
analyzer [Labmate-20, Invitron, Bangalore, India].

178 J. Preventive Cardiology Vol. 1 ■

■ No. 4 ■May 2012 J. Preventive Cardiology Vol. 1 ■
■ No. 4 ■ May 2012 179
 Mondal S, et al Tulsi and CVD

from 4600 mg/kg/bw (body weight) to 6200 mg/kg/bw in were the inclusion criteria for the enrollment of healthy Statistical analysis of results significant (intrasubject p=0.057) changes was seen in
5, 6
experimental mice. volunteers; a) age 18-60 years, either sex, b) not on any Tulsi extract treated groups during pre and post
regular medication and c) devoid of any medication at least Two-way ANOVA (Analysis of Variance) for crossover intervention periods. However, normally the fasting blood
Tulsi extract has been found to be cardioprotective in during last one month. Subjects suffering from different design was used to test the statistical significance of the glucose levels does not remain constant and it varies with
experimental animal studies. Long term oral diseases/disorders and/or having any kind of allergy, results. Independent ‘t-test’ was applied to compare the durations of overnight fasting, type of last meal taken along
administration of fresh Tulsi leaves augments cardiac undergone surgery during last one year, organ transplant baseline of two groups. Wilcoxon singed-rank test was with several other factors. Considering the dynamic
endogenous antioxidants and prevents myocardial necrosis recipients, chronic smokers, pregnant and lactating women applied to analyze triglycerides levels. The results were behavior of blood glucose levels under physiological
models in rats.7-9 Suanarunsawat et al (2011),10 conducted were excluded from the study. Enrolled individuals were considered significant if the p-value was 0.05 for period conditions this 1.56 mg/dl change may be ignored
animal experiments to investigate the lipid-lowering and randomized into two groups. Subjects and the staff directly effects and intervention effects. However, p-value of 0.10 clinically.
11
antioxidative activities of Ocimum sanctum L. (OS) leaf involved with the study subjects and data analysis were for carryover effects was considered significant. All
extracts in liver and heart of rats fed with high-cholesterol blinded about the interventional capsules. Allocation of statistical analysis was carried out using Stata 9.0 software Liver Function Tests (LFT) and Renal Function Tests
(HC) diet for seven weeks. group was concealed in opaque envelope. (Statacorp, Texas, USA). The blinding was decoded only (RFT)
after the statistical analysis.
Their results showed that OS suppressed the high levels of Tulsi extract and placebo administration Liver function tests were monitored by measuring SGOT
serum lipid profile and hepatic lipid content without ■
Results [Serum glutamate oxaloacetate transferase], SGPT [Serum
significant effects on fecal lipid excretion. Fecal bile acids Ethanolic (70%) extract of Ocimum sanctum Linn. leaves; glutamate pyruvate transferase] and also total bilirubin
excretion was increased in HC rats treated with OS. The ‘Tulsi extract’ was supplied by the Central Council for The volunteers of two groups were comparable on basic levels of serum in few subjects. It was found that there were
high serum levels of TBARS [Thiobarbituric Acid Research in Ayurveda and Siddha, Department of AYUSH, anthropometric and physiological parameters. We had a no significant differences during pre and post intervention
Reactive Substances] as well as AST [Aspartate Ministry of Family Health and Welfare, Government of wide range of age in inclusion criteria but subjects in periods in either group. There were no significant changes
Aminotransferase], ALT [Alanine Aminotransferase], AP India. Placebo (sucrose, nutrition grade) was supplied by relatively young age ranging from 22 to 37 years in the blood urea nitrogen (p=0.511) serum protein
[Alkaline Phosphatase], LDH [Lactate Dehydrogenase], the Dabur Pharmaceutical (India) Ltd, Ghaziabad (U.P.), participated in the study. The subjects had a healthy BMI (p=0.838) and creatinine (p=0.766) levels in either the
CK-MB [Creatine Kinase MB subunit] significantly India. Capsulation facilities were provided by the Dabur ranging from 17.3 to 25.64 kg/m2. Clinical biochemistry placebo or the Tulsi extract groups (Table 2).
decreased in HC rats treated with OS. OS treatment also Pharmaceutical to make 300 mg capsules using Tulsi parameters of the subjects were monitored at both pre and
suppressed the high level of TBARS and raised the low extract or placebo. Packaging of both, Tulsi extract and post intervention periods during the study. Total
activities of GPx [Glutathione Peroxides] and CAT placebo, were similar to avoid identification. The cholesterol (TC), triglycerides (TG), fasting blood glucose Triglycerides and total Cholesterol
[Catalase] without any impact on SOD [Superoxide intervention was allocated as per the randomized sequence levels (FBG), liver function test, renal function tests were
Dismutase] in the liver. Histopathological results also once the volunteer met the inclusion criteria and given a recorded. Table 1 shows the comparison of basic and There was no significant (p=0.128 & p=0.232) difference
confirmed that OS preserved the liver and myocardial written informed consent. Total period of trial was of 11 biochemical parameters of placebo and Tulsi extract at the baseline in triglycerides and cholesterol in both the
tissues. Present study was undertaken to validate the weeks in which two active administration period of 4 intervention groups at baseline. groups respectively (Table 2, 3). However it was found that
traditional claims of Tulsi regarding its usefulness in weeks each separated with a three weeks washout period in few individual who otherwise had an apparently normal
general health protection and to monitor various between. One capsule [code named, having Tulsi extract or Independent t-test applied. medical history had high cholesterol and triglycerides
biochemical parameters of healthy volunteers after placebo] of 300 mg was administered once daily, as per levels. It was observed that there was a significant decrease
administration of 300 mg of Tulsi extract or placebo on randomization, on empty stomach for four weeks followed Fasting blood glucose in the elevated levels of cholesterol (intra-subject p=0.003)
empty stomach for a period of 4 weeks. by a wash out period of three weeks before the volunteer of these individuals (n=6) in the Tulsi extract group and this
was crossover for the next intervention. There was a difference of about 1.56 mg/dl decrease in the decrease did not continue when subjects crossed-over to
■
Material and methods Tulsi extract treated groups (Table 2). When two-way the placebo group.
Collection of blood samples ANOVA for crossover design was applied a statistically
Design of the study
Venous blood samples of 5 ml were collected at four
Table 1: Comparison of Anthropometric and Physiological Parameters at
This was a double blind randomized controlled trial of total different time points i.e. i) before administration of
Baseline (before administration of capsules)
11 weeks in a crossover format which included a 3 weeks capsule, ii) after administration of placebo or Tulsi extract
wash-out period, to study the effects of ethanolic extract of capsules for four weeks iii) after the end of three weeks Basic parameters Baseline of Placebo-Tulsi Baseline of Tulsi- Placebo P-Value
Tulsi leaf on clinical biochemistry parameters of healthy washout period and finally iv) after completion of four sequence (n=12) sequence (n=10)
volunteers. The study was approved by the institutional weeks in crossover intervention period. The sampling was Age (years) 27.5 ±
4.75 (22-37) 26.5 ±
3.43 (23-35) 0.585
ethics committee on research involving human subjects done in fasting state [at least 12h] between 8:30 am to 10:00
and the trial was registered with Clinical Trial Registry of am at Department of Physiology, AIIMS, New Delhi. Height (cm) 166.16 ±
7.62 (153-180) 166.1 ±
5.40 (157-176) 0.981
India (No. CTRI/2009/091/000350). Subjects were contacted personally or telephonically twice Weight (kg) 62.5 ±
10.52 (50-80) 65.8 ±
6.05(54-78) 0.391
a week for better compliance and unused capsules were
2
Screening of Volunteers taken back from the subjects. Two volunteers did not BMI (kg/m ) 22.36 ±
2.52(17.3-25.43) 23.80 ±
1.13 (21.91-25.64) 0.111
continue the intervention due to their personal choice. Systolic BP (mm Hg) 109.16 ±
7.50 (100-120) 113.2 ±
7.37 (100-124) 0.220
Forty five volunteers were screened to enroll in the study. Clinical biochemical parameters were studied using
Twenty four healthy volunteers, who met the inclusion commercially available kits [Transasia, India; Randox, Diastolic BP (mm Hg) 74.16 ±
4.62 (68-80) 76.2 ±
3.45 (70-80) 0.265
criteria and gave their consent, were enrolled. Following United Kingdom] and semi-automatic biochemistry All values presented in mean ±
SD (range).P-value £
0.05 was considered as significant.
analyzer [Labmate-20, Invitron, Bangalore, India].

178 J. Preventive Cardiology Vol. 1 ■

■ No. 4 ■May 2012 J. Preventive Cardiology Vol. 1 ■
■ No. 4 ■ May 2012 179
 Mondal S, et al Tulsi and CVD

according to Jones and Kenward (2003)11, second 4. Mondal S, Mirdha BR, Mahapatra SC. The science behind
Table 2: Intervention Effects of Tulsi Extract or Placebo (Sucrose) Capsules on Biochemical sacredness of Tulsi (Ocimum sanctum Linn). Indian J Physio
intervention was not taken into consideration and results Pharmacol, 2009; 53: 291-302.
Parameters After 4 Weeks were analyzed based on first intervention only. When TGs 5. Bhargava KP, Singh N. Antistress activity of Ocimum sanctum Lin.
Biochemical Sequence of Baseline 4 weeks after 4 weeks after Baseline Period Carryover Intervention levels were analyzed, it was found that the reduction was Indian J Med. Res, 1981; 73: 443-451.
statistically not significant (Table 3). However, it was 6. Devi PU, Ganasoundari A. Radioprotective effect of leaf extract of
parameters interventions first second comparison effects effects effects Indian medicinal plant Ocimum sanctum. Indian J Exp Biol, 1995;
intervention intervention noticed that TGs levels were reduced in few subjects who 33:205-8.
had elevated levels. Thus, though not significant 7. Sharma M, Kishore K, Gupta SK, Joshi S, Arya DS.
First Second (at 0 week) (4th week) (11th Week) P-value# P-valuea P-valueb P-valuec statistically, a reduction trend was observed in persons with Cardioprotective potential of Ocimum sanctum in isoproterenol
induced myocardial infarction in rats. Mol Cell Biochem, 2001;
higher than the normal physiological reference ranges of 225(9): 75-83.
Fasting P TE 99.25 ±
3.84 99.58 ±
4.39 96.33 ±
3.60
blood (n=12)
TGs (Table 3). Therefore, the observed reduction in 8. Sood S, Narang D, Dinda, AK, Maulik SK. Chronic oral
glucose 0.546 0.109 0.940 0.0570 elevated levels of TGs and cholesterol in Tulsi extract administration of Ocimum sanctum Linn. augments cardiac
TE P 98 ±
3.85 97.7 ±
3.83 98 ±
3.85 treated group was due to treatment only. Earlier studies endogenous antioxidants and prevents isoproterenol-induced
(mg/dl) myocardial necrosis in rats. J Pharmac Pharmacol, 2005; 57: 127-
(n=10) have noted significant reduction in triglycerides, LDL [low 133.
Cholesterol P TE 187 ±
48.04 188.91 ±
44.97 174 ±
16.49 density lipoprotein] and VLDL [very low density 9. Arya D S, Nandave M, Ojha S K, Kumari S, Joshi S, Mohanty I.
lipoprotein] in the clinical trial of Tulsi leaves on NIDDM Myocardial salvaging effects of Ocimum sanctum in experimental
(mg/dl) 0.232 0.123 0.827 0.003* model of myocardial necrosis: a haemodynamic, biochemical and
TE P 206.18 ±
42.76 165.25 ±
12.92 200.6 ±
42.76 [Non-insulin dependent diabetes mellitus] patients.13,14 histoarchitectural assessment. Current Science, 2006; 91: 10
Animal studies had also showed that feeding of Tulsi leaves 667-672.
Total P TE 7.34 ±
0.76 7.59 ±
0.63 7.27 ±
0.79 15
reduces cholesterol levels. Present study also in consent 10. Suanarunsawat T, Ayutthaya WD, Songsak T, Thirawarapan S,
protein 0.515 0.711 0.608 0.838 Poungshompoo S. Lipid-lowering and antioxidative activities of
(g/ml) TE P 7.16 ±
0.54 7±
0.52 7.163 ±
0.54 with the findings of these studies. aqueous extracts of Ocimum sanctum L. leaves in rats fed with a
high-cholesterol diet. Oxid Med Cell Longev, 2011;
Blood Urea P TE 22 ±
5.93 22.25 ±
5.70 21.91 ±
5.5 Higher levels of TGs and cholesterol are one of the risk doi:10.1155/2011/962025.
Nitrogen factors of CAD in humans.
16,17
Thus, reduction in 11. Jones B, Kenward MG. Design and analysis of cross-over trials.
0.365 0.406 0.555 0.511
(mg/dl) TE P 19.08 ±
2.48 19.8 ±
2.484 21.9 ±
7.60 2nd Ed. Chapman & Hall/CRC. New York. US. 2003.
cholesterol and TGs seen in our study population is a very 12. Singh S, Majumdar DK, Yadav MR. Chemical and
Creatinine P TE 0.84 ±
0.25 0.82 ±
0.22 0.83 ±
0.21 positive outcome for reduction of dyslipidemia related pharmacological studies of Ocimum sanctum fixed oil. Indian J
(mg/dl) 0.780 0.139 0.819 0.766 CVD risks. This reinforces the traditional claim18 that Tulsi Exp Biol, 1996; 34: 1212-1215.
TE P 0.85 ±
0.07 0.83 ±
0.12 0.85 ±
0.09 13. Rai V, Mani UV, Iyer UM. Effect of Ocimum sanctum leaf powder
is good for heart. However caution should be taken in to on blood lipoproteins, glycated protein and total amino acids in
consideration while translating these findings in patients patients with non-insulin-dependent diabetes mellitus. J Nutr
All values presented in mean ± SD. P- Placebo, TE- Tulsi extract, #independent t-tests used. abcTwo-way ANOVA for crossover design. that our study population was of normal healthy person Environ Med. 1997;7:113–118.
#ac b 14. Agrawal P, Rai V, Singh RB. Randomized placebo-controlled,
p-Value £ 0. 05 and P-value £0. 10 were considered as significant. *Significant decrease [Intra-subject, n=6]. with no history of hypercholesterolemia or hyper- single blind trial of holy basil leaves in patients with noninsulin-
triglyceriglycemia. Thus, in patients with history of dependent diabetes mellitus. Int J Clin Pharmacol Ther.
■
Discussion three weeks was sufficient to undo the effects of first hypercholesterolemia or hyper-triglyceriglycemia this 1996;34:406–409.
intervention. property may be further explored. 15. Gupta S, Mediratta PK, Singh S, Sharma KK, Shukla R.
Antidiabetic, antihypocholestrolaemic and antioxidant effect of
All the subjects were relatively young, average age of both Ocimum sanctum (Linn) seed oil. Indian J Exp Biol, 2006; 44: 300-
groups were 27 ± 4.09 years, ranging from 22-37 years. Tulsi is a non-toxic plant and its LD50 value is very high5,6,12 ■
References 304.
They had a normal BMI, 23.08 ± 3.65 (ranging from 17.3 - however, for any side effects of Tulsi, biochemical 16. Gupta S, Mediratta PK, Singh S, Sharma KK, Shukla R.
2 1. Wealth of India. CSIR, Publication and information directorate, Antidiabetic, antihypocholestrolaemic and antioxidant effect of
25.43 kg/m ). There was no significant period or carryover parameters were evaluated. It was observed that Ocimum sanctum (Linn) seed oil. Indian J Exp Biol, 2006; 44: 300-
intervention with 300 mg capsules of Tulsi extract for 28 New Delhi, 1991; VII: 79-89.
effects observed (Table 2).Thus, the washout period of 2. WHO Monogram on selected medicinal plants. Dr Xiaorui Zhang, 304.
days on empty stomach did not show any toxic effects as Acting Coordinator, Traditional Medicine, Department of Essential 17. Talayero BG, Sacks FM. The role of triglycerides in
evidenced by basic biochemical results. There were no Drugs and Medicines Policy, World Health Organization. Geneva, atherosclerosis. Curr Cardiol Rep. 2011; 13(6):544-52.
Table 3: Effects of Tulsi Extract or Placebo significant changes observed in BMI, BP, fasting blood 2002; Vol 2: 206- 216. 18. Saggini A, Anogeianaki A, Angelucci D, et al. Cholesterol and
(Sucrose) Capsules on Triglycerides After 3. Gupta SK, Jai Prakash and Srivastava S. Validation of traditional vitamins: revisited study. J Biol Regul Homeost Agents. 2011;
sugar, LFT and RFT. However, some of the subjects (n=6) claim of Tulsi, Ocimum sanctum Linn. as a medicinal plant. Indian 25(4):505-15.
4 Weeks of Intervention who had higher than the normal physiological ranges of J Exp Biol, 2002; 40: 765-773.
Address for correspondence
Triglycerides (mg/dl) cholesterol showed significant reduction (intra-subject, p= Dr. Sushil Chandra Mahapatra: Email: scmahapatra@gmail.com
Baseline After 4 weeks of 0.003) in the elevated levels after taking capsules of Tulsi
intervention extract for 4 weeks. However, no significant reduction was
observed in the subjects who had cholesterol levels within
Placebo (n=12) 57.5 (25-194) 61.5 (39-190) normal physiological reference ranges.
Tulsi extract (10) 78.5 (25-350) 74 (31-282)
P-value 0.128 0.350 The reduction were also observed in the subjects who had
higher than the normal physiological ranges of TGs levels,
Value presented in median (range). P-value 0.05 was considered as
significant. Wilcoxon singed-rank test applied.
however when tested for normal distribution of the data, it
showed the carryover effects after washout period. Thus,

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No. 4 ■ May 2012 181
 Mondal S, et al Tulsi and CVD

according to Jones and Kenward (2003)11, second 4. Mondal S, Mirdha BR, Mahapatra SC. The science behind
Table 2: Intervention Effects of Tulsi Extract or Placebo (Sucrose) Capsules on Biochemical sacredness of Tulsi (Ocimum sanctum Linn). Indian J Physio
intervention was not taken into consideration and results Pharmacol, 2009; 53: 291-302.
Parameters After 4 Weeks were analyzed based on first intervention only. When TGs 5. Bhargava KP, Singh N. Antistress activity of Ocimum sanctum Lin.
Biochemical Sequence of Baseline 4 weeks after 4 weeks after Baseline Period Carryover Intervention levels were analyzed, it was found that the reduction was Indian J Med. Res, 1981; 73: 443-451.
statistically not significant (Table 3). However, it was 6. Devi PU, Ganasoundari A. Radioprotective effect of leaf extract of
parameters interventions first second comparison effects effects effects Indian medicinal plant Ocimum sanctum. Indian J Exp Biol, 1995;
intervention intervention noticed that TGs levels were reduced in few subjects who 33:205-8.
had elevated levels. Thus, though not significant 7. Sharma M, Kishore K, Gupta SK, Joshi S, Arya DS.
First Second (at 0 week) (4th week) (11th Week) P-value# P-valuea P-valueb P-valuec statistically, a reduction trend was observed in persons with Cardioprotective potential of Ocimum sanctum in isoproterenol
induced myocardial infarction in rats. Mol Cell Biochem, 2001;
higher than the normal physiological reference ranges of 225(9): 75-83.
Fasting P TE 99.25 ±
3.84 99.58 ±
4.39 96.33 ±
3.60
blood (n=12)
TGs (Table 3). Therefore, the observed reduction in 8. Sood S, Narang D, Dinda, AK, Maulik SK. Chronic oral
glucose 0.546 0.109 0.940 0.0570 elevated levels of TGs and cholesterol in Tulsi extract administration of Ocimum sanctum Linn. augments cardiac
TE P 98 ±
3.85 97.7 ±
3.83 98 ±
3.85 treated group was due to treatment only. Earlier studies endogenous antioxidants and prevents isoproterenol-induced
(mg/dl) myocardial necrosis in rats. J Pharmac Pharmacol, 2005; 57: 127-
(n=10) have noted significant reduction in triglycerides, LDL [low 133.
Cholesterol P TE 187 ±
48.04 188.91 ±
44.97 174 ±
16.49 density lipoprotein] and VLDL [very low density 9. Arya D S, Nandave M, Ojha S K, Kumari S, Joshi S, Mohanty I.
lipoprotein] in the clinical trial of Tulsi leaves on NIDDM Myocardial salvaging effects of Ocimum sanctum in experimental
(mg/dl) 0.232 0.123 0.827 0.003* model of myocardial necrosis: a haemodynamic, biochemical and
TE P 206.18 ±
42.76 165.25 ±
12.92 200.6 ±
42.76 [Non-insulin dependent diabetes mellitus] patients.13,14 histoarchitectural assessment. Current Science, 2006; 91: 10
Animal studies had also showed that feeding of Tulsi leaves 667-672.
Total P TE 7.34 ±
0.76 7.59 ±
0.63 7.27 ±
0.79 15
reduces cholesterol levels. Present study also in consent 10. Suanarunsawat T, Ayutthaya WD, Songsak T, Thirawarapan S,
protein 0.515 0.711 0.608 0.838 Poungshompoo S. Lipid-lowering and antioxidative activities of
(g/ml) TE P 7.16 ±
0.54 7±
0.52 7.163 ±
0.54 with the findings of these studies. aqueous extracts of Ocimum sanctum L. leaves in rats fed with a
high-cholesterol diet. Oxid Med Cell Longev, 2011;
Blood Urea P TE 22 ±
5.93 22.25 ±
5.70 21.91 ±
5.5 Higher levels of TGs and cholesterol are one of the risk doi:10.1155/2011/962025.
Nitrogen factors of CAD in humans.
15,16
Thus, reduction in 11. Jones B, Kenward MG. Design and analysis of cross-over trials.
0.365 0.406 0.555 0.511
(mg/dl) TE P 19.08 ±
2.48 19.8 ±
2.484 21.9 ±
7.60 2nd Ed. Chapman & Hall/CRC. New York. US. 2003.
cholesterol and TGs seen in our study population is a very 12. Singh S, Majumdar DK, Yadav MR. Chemical and
Creatinine P TE 0.84 ±
0.25 0.82 ±
0.22 0.83 ±
0.21 positive outcome for reduction of dyslipidemia related pharmacological studies of Ocimum sanctum fixed oil. Indian J
(mg/dl) 0.780 0.139 0.819 0.766 CVD risks. This reinforces the traditional claim17 that Tulsi Exp Biol, 1996; 34: 1212-1215.
TE P 0.85 ±
0.07 0.83 ±
0.12 0.85 ±
0.09 13. Rai V, Mani UV, Iyer UM. Effect of Ocimum sanctum leaf powder
is good for heart. However caution should be taken in to on blood lipoproteins, glycated protein and total amino acids in
consideration while translating these findings in patients patients with non-insulin-dependent diabetes mellitus. J Nutr
All values presented in mean ± SD. P- Placebo, TE- Tulsi extract, #independent t-tests used. abcTwo-way ANOVA for crossover design. that our study population was of normal healthy person Environ Med. 1997;7:113–118.
#ac b 14. Agrawal P, Rai V, Singh RB. Randomized placebo-controlled,
p-Value £ 0. 05 and P-value £0. 10 were considered as significant. *Significant decrease [Intra-subject, n=6]. with no history of hypercholesterolemia or hyper- single blind trial of holy basil leaves in patients with noninsulin-
triglyceriglycemia. Thus, in patients with history of dependent diabetes mellitus. Int J Clin Pharmacol Ther.
■
Discussion three weeks was sufficient to undo the effects of first hypercholesterolemia or hyper-triglyceriglycemia this 1996;34:406–409.
intervention. property may be further explored. 15. Gupta S, Mediratta PK, Singh S, Sharma KK, Shukla R.
Antidiabetic, antihypocholestrolaemic and antioxidant effect of
All the subjects were relatively young, average age of both Ocimum sanctum (Linn) seed oil. Indian J Exp Biol, 2006; 44: 300-
groups were 27 ± 4.09 years, ranging from 22-37 years. Tulsi is a non-toxic plant and its LD50 value is very high5,6,12 ■
References 304.
They had a normal BMI, 23.08 ± 3.65 (ranging from 17.3 - however, for any side effects of Tulsi, biochemical 16. Talayero BG, Sacks FM. The role of triglycerides in
2 1. Wealth of India. CSIR, Publication and information directorate, atherosclerosis. Curr Cardiol Rep. 2011; 13(6):544-52.
25.43 kg/m ). There was no significant period or carryover parameters were evaluated. It was observed that 17. Saggini A, Anogeianaki A, Angelucci D, et al. Cholesterol and
intervention with 300 mg capsules of Tulsi extract for 28 New Delhi, 1991; VII: 79-89.
effects observed (Table 2).Thus, the washout period of 2. WHO Monogram on selected medicinal plants. Dr Xiaorui Zhang, vitamins: revisited study. J Biol Regul Homeost Agents. 2011;
days on empty stomach did not show any toxic effects as Acting Coordinator, Traditional Medicine, Department of Essential 25(4):505-15.
evidenced by basic biochemical results. There were no Drugs and Medicines Policy, World Health Organization. Geneva,
Table 3: Effects of Tulsi Extract or Placebo significant changes observed in BMI, BP, fasting blood 2002; Vol 2: 206- 216. Address for correspondence
(Sucrose) Capsules on Triglycerides After 3. Gupta SK, Jai Prakash and Srivastava S. Validation of traditional Dr. Sushil Chandra Mahapatra: Email: scmahapatra@gmail.com
sugar, LFT and RFT. However, some of the subjects (n=6) claim of Tulsi, Ocimum sanctum Linn. as a medicinal plant. Indian
4 Weeks of Intervention who had higher than the normal physiological ranges of J Exp Biol, 2002; 40: 765-773.
Triglycerides (mg/dl) cholesterol showed significant reduction (intra-subject, p=
Baseline After 4 weeks of 0.003) in the elevated levels after taking capsules of Tulsi
intervention extract for 4 weeks. However, no significant reduction was
observed in the subjects who had cholesterol levels within
Placebo (n=12) 57.5 (25-194) 61.5 (39-190) normal physiological reference ranges.
Tulsi extract (10) 78.5 (25-350) 74 (31-282)
P-value 0.128 0.350 The reduction were also observed in the subjects who had
higher than the normal physiological ranges of TGs levels,
Value presented in median (range). P-value 0.05 was considered as
significant. Wilcoxon singed-rank test applied.
however when tested for normal distribution of the data, it
showed the carryover effects after washout period. Thus,

180 J. Preventive Cardiology ■ Vol. 1 ■

No. 4 ■ May 2012 J. Preventive Cardiology ■ Vol. 1 ■
No. 4 ■ May 2012 181