Study of the Antidiabetic Activity of Huernia Sp. Nov. aff. Boleana Growing in High Altitude Areas of Southwest Saudi Arabia

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1 ISSN: RESEARCH ARTICLE Annals of Biological Sciences 2015, 3 (4):15-20 Study of the Antidiabetic Activity of Huernia Sp. Nov. aff. Boleana Growing in High Altitude Areas of Southwest Saudi Arabia Saleh O. Alzahrani 1, Abdullah M. Alwagdani 1, Ahmed M. Alotaibi 1, Mohamed Ghazi Hamaidi 2, Abdelrahman Nasr 3, Mayyas Al-Remawi 2, Yaser G. Gouda 4, 5* and Khaled M. Mohamed 4, 5 1 6th year students, Department of Clinical Pharmacy, College of Pharmacy, Taif Univ., Taif, Al- Haweiah, P.O. Box 888, KSA 2 Department of Pharmaceutics and Pharmaceutical Technology, College of Pharmacy, Taif Univ., Taif, Al-Haweiah, P.O. Box 888, KSA 3 Department of Pharmacology and Toxicology, College of Pharmacy, Taif Univ., Taif, Al-Haweiah, P.O. Box 888, KSA 4 Department of Pharmacognosy, College of Pharmacy, Taif Univ., Taif, Al-Haweiah, P.O. Box 888, KSA 5 Departmentof Pharmacognosy, Faculty of Pharmacy, Assiut Univ., Assiut, 71526, Egypt * Corresponding ghallab68@yahoo.com ABSTRACT In this study a new plant species known as Huernia Sp. Nov. aff. Boleana (Apocyanaceae, formerly known as Asclepiadaceae) growing in high altitude Southwest areas of KSA have been investigated for its use as antidiabetic. Both total methanolic and chloroform extracts showed significant hypoglycemic activity over all time intervals for STZ diabetic rats compared to the control group. Keywords: Huernia Sp. Nov. aff. Boleana, Apocyanaceae, Antidiabetic. INTRODUCTION Diabetes is a common disease that affects glucose metabolism and results in high blood glucose level over a long period. Many antidiabetic agents were used to achieve normal glycemic level [1].Far from chemical drugs, different ways for controlling glucose level have been investigated such as low sugar diets and exercise [2]. However the search for better and safer ways of reducing blood glucose level throughout human history led to the use of herbs as antidiabetic medicines such as cinnamon and fenugreek [3]. One interesting plant that has been used in folk medicine as antidiabetic is Huernia Sp. Nov. aff. Boleana belongs to Apocynaceae family (formally known as Asclepiadaceae). It is a wild plant grows on mountains of south west region of Kingdom of Saudi Arabia and known locally as (Zaidah). Asclepiadaceae now is treated as a subfamily (Asclepiadoideae) in the Apocynaceae which has about 348 genera and 20 species [4]. Many plants of Asclepiadoideae were reported to have antidiabetic activity as Gymnema sylvestre[5]. Other plants belong to the same subfamily such as Marsdenia tenacissima[6], Pergularia daemia[7], Hemidesmus indicus[8], Caralluma umbellate [9] and Cryptolepis buchanani[10] are used in folkloric medicine to treat cough, asthma, some types of cancer, liver disorders, and as anti-nociceptive and antiinflammatory herbal drugs [6-10]. Genus Huernia comprises about 44 species [11] with a little knowledge about their medicinal uses and biological activities. It was previously reported the acetylcholinesterase inhibition, antioxidant, anti-inflammatory, antimicrobial and phytochemical properties of Huernia hystrix [12]. 15

2 In the present study a new species known as Huernia Sp. Nov. aff. Boleana grows on mountains of south west region of Kingdom of Saudi Arabia has been investigated for its use as antidiabetic based on the finding of Asclepiadoideae and the traditional use of the Huernia sp. Nov. aff. Boleana in the folkloric medicine. According to the best of our knowledge, no data could be traced about the pharmacognostical, phytochemical or biological studies on this species. MATERIALS AND METHODS Plant materials The plant material of Huerina Sp. Nov. aff. Boleana was collected from Rusaba, southern west region, Kingdom of Saudi Arabia during March The Plant was kindly identified by Prof. Dr. Yassin M. Al-Sodany, Botany Dept., College of Science, Taif University, Taif, Saudi Arabia. A voucher specimen was deposited at the Herbarium of Pharmacognosy department, College of Pharmacy, Taif University, Taif, KSA. Standard materials Methanol, n-hexane, Chloroform and Streptozotocin were purchased from Sigma Aldrich (USA). Glucose level was monitored using Glucometer (Bionime GM300, USA Corp., USA). Preparation of the crude plant extracts The air-dried powdered plant of Huerina Sp. Nov. aff. Boleana (4g) was extracted with methanol till exhaustion. The total methanolic extract was concentrated under vacuum using rotary evaporator to give dried total methanolic extract (87g).Forty seven grams of the total methanolic extract was suspended in a little amount of distilled water and successively fractionated with n-hexane then chloroform to afford dried n-hexane fraction (3.4g), chloroform fraction (23.8g) and the aqueous fraction which was lyophilized giving (16.4g). All fractions were kept in refrigerator until used. Animals Wistar male rats ( g) were obtained from the animal house of King Abdul-Aziz University, Jeddah, KSA. The rats were housed in standard environmental conditions and fed standard laboratory diet with free access to water and kept for one day with 12 h light/dark cycle prior conducting the anti-diabetic study. All applicable institutional and/or national guidelines for the care and use of animals were followed. The animal studies were carried out according to the European Community Council Directives published in November 24, 1986 (86/9/EEC). Anti-diabetic activity test Streptozotocin (STZ, 1 mg/dl) was given intraperitoneally to the tested rats. After 1 week, rats having a glucose level more than 250 mg/dl were selected for the study. Animals were classified into five groups (each of six rats). The total methanolic extract, n-hexane, chloroform and aqueous fractions and distilled water as (control group) were given orally at a dose of 2ml (75 mg/kg body weight) as a preliminary screening dose using stomach tube. Statistical analysis The statistical significances were examined using analysis of variance one way ANOVA. All statistical analysis was conducted via (SPSS program version ) MS Excel The significant was considered when p-value less than Table 1: The antidiabetic activity of total methanolic, n-hexane, chloroform and aqueous extracts of Huerina Sp. Nov. aff. Boleana: Mean SE % Mean Total SE % Mean n-hexane SE % Mean Chloroform SE % Mean Aqueous SE % Mean: Arithmetic mean, SE: Standard error 16

3 Hexane Extract Figure 1: Glucose level of STZ-induced diabetic rats given an oral dose (75 mg extract/kg) of Huernia n-hexane extract and total extract compared to control Chloroform Extract Figure 2: Glucose level of STZ-induced diabetic rats given an oral dose (75 mg extract/kg) of Huernia chloroform extract and total extract compared to control. 17

4 Aqueous Extract Figure 3: Glucose level of STZ-induced diabetic rats given an oral dose (75 mg extract/kg) of Huernia aqueous extract and total extract compared to control Average Figure 4: Glucose level of STZ-induced diabetic rats given an oral dose (75 mg extract/kg) of Huernia n-hexane extract, chloroform extract, aqueous extract (average) and total extract compared to control RESULTS AND DISCUSSION Figures 1-3 and table 1 showed the percent change in glucose level of Huerina Sp. Nov. aff. Boleana extracts given to STZ-diabetic rats on fasted state compared to a control group. The obtained results indicated high hypoglycemic activity of all extracts compared to the values obtained by the control. 18

5 As shown in figure 1 and table 1, the total methanolic extract showed significant hypoglycemic activity over all time intervals (8 hours, one hour interval).the blood glucose level were lowered by 8.50%, 4.27%, 11.33%, 9.19%, 25.57%, 24.71%, 23.73% and 17.36%respectively(p<0.05) compared to the initial. Also, the n-hexane extract showed significant hypoglycemic activity nearly over all time intervals except after 2 hours where the level was increased by 4.18%. The percent decrease in blood glucose level was2.35%, 12.72%, 5.21%, 12.85%, 5.25%, 4.41% and 11.74%after 1,3,4,5,6,7,8 hours respectively (p<0.05)compared to the initial. From the above data, the total methanolic extract showed higher decrease in glucose level compared to that of n-hexane extract at nearly all time intervals. The chloroform extract showed significant hypoglycemic activity (p<0.05) over all time intervals for STZ induced diabetic rats compared to the initial (figure 2 and table 1,)and the decrease of blood glucose level were14.52%, 19.53%, 22.19%, 26.31%, 23.98%, 29.17%, 30.88% and 19.75% respectively. The chloroform showed higher decrease in glucose level than the total methanol extract after 2, 3 and 4 hours. As shown in figure 3 and table 1,aqueous extract revealed significant hypoglycemic activity nearly over all time intervals except the last hour (increase by 2.69%) and the percent decrease in blood glucose level was3.08%, 9.15%, 4.00%, 10.92%, 14.08%, 15.96%and 13.67% respectively (p<0.05) compared to the initial. The total methanolic extract showed higher decrease in glucose level than the aqueous extract (p<0.05) at nearly all time intervals. The mathematical average of hypoglycemic effect of all extracts was compared to the total extract as shown in figure 4. It was obvious that the plots were almost similar (p>0.05). Thus, ensures that the finding of hypoglycemic activity of total methanolic extract was much related to that of the mathematical average for all portions. The antidiabetic activity of herbal drugs is attributed to both polar and non-polar compounds [13, 14]. In case of Huerina Sp. Nov. aff. Boleana the activity is expected to be for both non-polar and less polar compounds which are present in n-hexane and chloroform fractions. Similar findings were observed for different plants having hypoglycemic activity; for example, a study on Malva parviflora leaves indicated the hypoglycemic activity of hexane extract using STZ-induced diabetic rat model. The study showed that the best hypoglycemic activity came from an extremely hydrophobic solvent rather than chloroform or methanol. This confirms the hydrophobic nature of the therapeutic active components and their pharmacological activities [13].However another study showed the preference of hydrophilic extracts on activity of the methanolic extract of Caralluma tuberculata as well as its chloroform, n-butanol and the remaining water fractions in STZ-induced diabetic rats [14]. Both methanolic extract and water fractions showed high potency, however methanolic extract had superior activity [14]. Investigation of the hypoglycemic activity of Ficus deltoidea leaves extracts (methanol, n-hexane, chloroform, and n-butanol) showed that the methanol extract exhibited higher blood glucose lowering activity than n-butanol extract[15]. Further studies will be done on this plant including the isolation and structure elucidation of chemical component(s) responsible for this activity and other activities. REFERENCES [1] American Diabetes Association, Diabetes Care. Diagnosis And Classification Of Diabetes Mellitus, 2010, 33(Suppl 1): P. S62-S69. [2] E.J. Stephenson, W. Smiles, And J.A. Hawley, Med Sport Sci.2014,, [3] R. Deng, Recent Pat Food Nutr Agric,2012,4(1), 50-. [4] M.E. Endress And P.V. Bruyns, Bot. Review,2000,66(1), [5] S. Sathya, R. Kokilavani, K. Gurusamy,Ancient Science Of Life.2008,28(2): [6] B. Ye, J. Li, Z. Li, J. Yang, T. Niu And S. Wang, Journal Of Ethnopharmacology,2014,153(1), [7] S.V. Sureshkumar And S.H. Mishra, Journal Of Ethnopharmacology,2006,107(2), [8] P.R. Verma, A.A. Joharapurkar, V.A. Chatpalliwar And A.J. Asnani, Journal Of Ethnopharmacology, 2005,102(2), [9] M. Ramesh, Y.N. Rao, M.R. Kumar, M.C. Prabhakar And B.M. Reddy, Journal Of Ethnopharmacology, 1999,68, [10] P. Laupattarakasem, T. Wangsrimongkol, R. Surarit And C. Hahnvajanawong, Journal Of Ethnopharmacology, 2006,108(3), [11] Wikipedia, The Free Encyclopedia. [12] S.O. Amoo, J.F. Finnie, And J.V. Staden, Phytotherapy Research,2012,26(5), [13] P. Gutierrez, Food Funct.,2012, 3(4), [14] E.A. Abdel-Sattar, H.M. Abdallah, A. Khedr, A.B. Abdel-Naim And I.A. Shehata, Food Chem Toxicol,2013,59,

6 [15] Y. Ilyanie, T.W. Wong, And C.Y. Choo, J Complement Integr Med,2011,8,