Egyptian Dermatology Online Journal, Volume 9 Number 2
EDOJ



Contents






Web
www.edoj.org.eg



Effect of ethanolic fraction of Hibiscus rosa sinensis l., leaves in androgenic alopecia

Sukirti Upadhyay1, Prashant Upadhyay1, Reetesh Vinode2 and Vinod K Dixit2

1 School of Pharmaceutical Sciences, IFTM University, Moradabad-244001, UP, India.

2 Department of Pharmaceutical Sciences, Dr H S Gaur University Sagar -470003.M P, India.

Egyptian Dermatology Online Journal 9 (2): 5, December 2013

Corresponding Author: Sukirti Upadhyay
e-mail: sukirtiupadhyay@gmail.com

Submitted: 24 September 2013
Accepted: 23 November 2013
 
Click Here for a Printable PDF version  






Abstract

Hibiscus rosa sinensis leaves are well known as hair growth promoter in traditional and folklore medicines. In this study, experiments were performed on male albino rats for induction and development of androgenic alopecia by subcutaneous dose of testosterone for three weeks (21 days). Animals were divided into three groups and group one, two and three animals were treated with vehicle, finasteride and ethanolic extract of Hibiscus rosa sinensis leaves topically along with testosterone respectively. Animals were observed for signs of alopecia on dorsal skin. After the treatment period one animal from each group was euthanized and follicular morphology and density were studied. The animals treated with testosterone and vehicle become alopecic from the second week of treatment, while animals treated with finasteride and ethanolic extract of Hibiscus rosa sinensis did not become alopecic and follicular morphology study also supported this. We concluded that ethanolic extract of H.rosasinensis posses significant anti androgenic alopecia activity as proved by this study and it is comparable to standard FDA approved drug for androgenic alopecia finasteride.

Introduction

Alopecia has a great impact on a person's self-respect, mental health, and overall quality of life. The disorder, androgenic alopecia (AGA) is the most common type of hair loss, which affects large numbers of both men and women. [1] It affects at least half of all men by the age of 50, and up to 70% of 70-year-old men. [2] AGA is caused by excessive activity of the 5alpha reductase enzyme in hair follicles. [3] At present, there are some medicines that are used to treat AGA. For example, 5alpha reductase inhibitors, finasteride and dutasteride, are used to treat androgen-related disorders. [4] But these medicines have several undesirable side effects, for example, impotence (erectile dysfunction), abnormal ejaculation, decreased ejaculatory volume, abnormal sexual function, gynecomastia, testicular pain, impairment of muscle growth, and severe myopathy. [5] Alternatively some herbal extracts are found effective in androgenic alopecia such as that of Cuscuta reflexa [6], Abrus precatorius [7] and Carthamus tinctorius [8] etc. Out of praised plants for hair growth promotion, the herb Hibiscus rosa-sinensis L. (Malvaceae) is native to China but easily available in various part of India, was found to be a promoter of hair growth and its leaves' extract is more potent than the flowers' extract. [9] As The herb is a source of antioxidants like Taraxeryl acetate, sterculic and malvalic acids, quercetin and its glycosides, cyanin and cyanidin chlorides, kaemperol-3-xylosylglucoside, thiamine, riboflavin, niacin and ascorbic acid. [9]

In the current study, we are evaluating potency of ethanolic extract of Hibisus rosa sinensis leaves on androgenic alopecia experiments, conducted in male albino rats. No previous work is reported for this aspect.

Material and Method

Plant material

Fresh leaves were collected from garden in Girls Hostel, U.T.D Sagar in the month of Feb 2008 and identified by Dr Pradeep Tiwari of Department of Botany. Dr H.S. Gaur University, Sagar, and a voucher specimen no. (Bot/H/2792) was preserved there for future references

Chemicals

Finasteride was procured from CIPLA LTD, Sikkim, India while petroleum ether, ethanol and propylene glycol were obtained from Central Drug House, Delhi, India. Testosterone is obtained from German Remedies, Mumbai, India.

Preparation of extracts

Leaves were dried in oven at 35- 37C and then it was defatted in petroleum ether(60-80)C in soxhlet apparatus then dried and macerated with ethyl alcohol overnight and then the filtrate was concentrated by vacuum under reduced pressure the yield was 30% w/w.

Animals

Male Wistar albino rats, weighing 120-150 g, 3-4 months old were used for hair growth studies. The rats were placed in cages and kept in standard animal house conditions in 12 h light and 12 h dark cycle, fed with standard pellet diet ad libitum and allowed free access to drinking water.

Testosterone test solution

Marketed preparation of testosterone Aquavirion was diluted up to 5 ml with water for injection. This was able to produce the concentration of 5mg/mL

Finasteride, extract solution

The 2% finasteride and extract solution was prepared in vehicle (ethanol/propylene glycol/water, r = 8: 1: 1).

Phytochemical Screening

Ethanolic root extract was screened by performing various microchemical tests for the presence of active phyto-constituents.

Toxicity studies

Toxicity studies of extract on skin of animals were carried out by the application of ethanolic extract in a concentration of up to 10% for seven days on the shaved denuded skin of albino rats. They were further observed for signs of toxicity such as swelling, itching, erythema etc. [10]

Treatment of animals

The animals were divided into three groups of six animals in each group. Group 1M, 2M and 3 M consists of male albino rats. [11] All animals were treated with 0.1 mL of testosterone daily for three weeks and group1M, 2M and 3M receives topical massage of vehicle, finasteride and ethanolic extract of leaf respectively.

Results and Discussion
 

Phytochemical screening

Hibiscus rosa sinensis leaf ethanolic extract shows presence of carbohydrates, flavonoids and phenolics.

Toxicity studies

The extract was considered safe for topical use as no signs of toxicity as erythrema, swellings, irritation were produced in rats for seven days study.

Effect of hair growth

The 1M group of animals on a dose of testosterone and vehicle become alopecic as hair shedding starts from day fourteen from the dorsal skin and alopecia progressed from the cranial to the caudal region while in groups 2M and 3M the animals did not develop signs of alopecia up to 21 days of the study. The finasteride and ethanolic extract treated animals (Fig 2) did not develop signs of alopecia and the skin section study shows higher follicle density, anagenic follicles while the vehicle treated animals developed alopecia (Fig 1) which shows that the vehicle does not have anti androgenic alopecia activity while finasteride and the extract successfully combat alopecia. The statistical study of hair follicles by occular micrometer also supports the observed results significantly (Fig 2, 3, 4). Skin section of the animal shows hair follicles in anagenic stage and very few follicle in telogenic stage. In figure 4 the column graph shows a high percentage of anagenic follicles in extract treated animals followed by finasteride treated animals while the animals on vehicle shows least percent of anagenic follicles. The quantitative study of hair follicle density also shows maximum hair density in extract treated animals. This activity in extract may be due to the presence of antioxidant potential of the plant or presence of flavonoids. [12] One of the major causes of androgenic alopecia is the reduced blood supply to the androgen sensitive and genetically predisposed hair follicles. However, a number of investigators have shown that polar compounds like flavonoids possess hair growth promoting activity by strengthening the capillary wall of the smaller blood vessels supplying the hair follicles; improve blood circulation to nourish the hair follicles and thereby promoting hair growth. [13,14,15] . As the extract is rich in flavonoid content, strengthening capillary wall may be the reason for increased blood circulation and combating the effect of androgen supplied by the subcutaneous route.




Fig 1: The dorsal skin of animal on dose of testosterone and vehicle (1M).

 




Fig 2: The dorsal skin of extract treated animal along with testosterone (3M) does not develop alopecia.

 




Fig 3: Anagen/ Telogen ratio of different groups of animals.

 




Fig 4: Effect of various treatment along with testosterone on Follicle density of male rats.

 

Conclusion

Ethanolic extract of leaves posses effect against androgenic alopecia in male rats so conclusively it may be used in the future as a hair restoring agent for males suffering from alopecia.

Acknowledgement

One of the authors SU is grateful to Prof MD Kharya, Head, Department of Pharmaceutical Sciences, Dr H. S. Gaur University, Sagar, for providing research facilities and UGC for fellowship.

References


1. Sinclair RD. Male androgenetic alopecia. Journal of Men's Health and Gender 2004; 1, 319-327.

2. Treb RM. Molecular mechanisms of androgenetic alopecia. Experimental Gerontology 2002; 37, 981-990.

3. Sawaya ME. Novel agents for the treatment of alopecia. Seminars in Cutaneous Medicine and Surgery 1998;17, 276-283.

4. Robinson A J DeLucca I Drummond S Boswell G A. Steroidal nitrone inhibitor of 5alphareductase. Tetrahedron Letter 2003; 44, 4801-4804.

5. Lacy CF Armstrong L L Goldman M P Lance L Drug Information Handbook with International Trade Names Index, 17th ed. Lexi .Comp Inc., United States, 2008; pp. 652-653.

6. Chauhan NS, Dixit VK Effect of Cuscuta reflexa Roxb on androgen-induced alopecia. J Cosmet Dermatol 2008; 7: 199-204.

7. Upadhyay S ,Ghosh A K , Singh V and Dixit V K Effect of petroleum ether and ethanol fractions of Abrus precatorius on androgenic alopecia. Revista Brasileira de armacognosia, 2012.22(2): 359-363.

8. Kumar N et al., 5 alpha reductase inhibition and hair growth promotion of some Thai plants traditionally used for hair treatment Journal of Ethnopharmacology 2012; 139 765- 771.

9. Adhirajan N, et al, In vivo and in vitro evaluation of hair growth potential of Hibiscus rosa-sinensis Linn. J Ethnopharmacol. 2003; 88(2-3):235-9.

10. ASTM Standard practice for testing biomaterials in rabbits for primary skin irritation, Philadelphia: American Society for Testing of Materials, F 1998; 719-781: 178-179.

11. Matias JR, Malloy V, Orentreich N. Animal models of androgen-dependent disorders of the pilosebaceous apparatus. 1. The androchronogenetic alopecia (AGA) mouse as a model for male-pattern baldness. Arch Dermatol Res 1989; 281: 247-253.

12. Divya MJ, Sowmia C, Dhanya KP, and Joona K 2. Screening of Antioxidant, Anticancer Activity and Phytochemicals in Methanolic Extract of Hibiscusrosa-Sinensis Leaf Extract. Research Journal of Pharmaceutical, Biological and Chemical Sciences 2013 ;4 (2):1308-16

13. Kobayashi N, Suzuki R, Koide C, Suzuki T, Matsuda H, et al. Effect of leaves of Ginkgo biloba on hair growth in C 3Hstrain mice. Yakugaku Zasshi.1993;113: 718-724.

14. Awe EO, Makinde JM . The hair growth promoting effect of Russelia equisetiformis (Schclect&Chan). J Natl Prods 2009; 2: 70-73.

15. Kawano M, Han J, Kchouk ME, Isoda H Hair growth regulation by the extract of aromatic plant Erica multiflora. J Nat Med 2009; 63: 335-339.

2013 Egyptian Dermatology Online Journal