Effect of Hot Aqueous Extract of Rheum Ribes Roots on Some Hormonal and Biochemical Parameters in Induced Polycystic Ovary Syndrome in Local Female Rabbits

EurAsian Journal of BioSciences Eurasia J Biosci 12, 419-423 (2018)

Effect of hot aqueous extract of Rheum ribes roots on some hormonal and biochemical parameters in induced Polycystic Ovary Syndrome in local female rabbits

Aseel Mokdad Hatam AbdulWahed 1, Salih M. Rahem Al-obaidi 2, Abdul-monaim H. M. Al-samarrai 3 1 Samarra university, College of applied sciences, Department of analysis pathological, IRAQ 2 Tikrit university, college of education and pure sciences, department of biology, IRAQ 3 Samarra university, College of education, Department of chemistry, IRAQ

Abstract The study aimed to evaluate the effect of aqueous extract of R. ribes roots and metformin in treatment of induced PCOS. 40 adult female rabbits (1000-1600g) use in this study, 30 rabbits injected with TP (100mg/Kg) for 4 days consecutive and left 3 days for developing the syndrome. The female rabbits divided to four groups: C+ve group, C-ve group, G1 (R. ribes extract 300mg/Kg) and G2 (metformin group 20mg/Kg). Treatment period was 30 days. The hormonal and biochemical assays include: AMH, Testosterone, Insulin, HOMA-IR and Glucose levels. The results showed significant increase at P≥0.05 in levels of AMH, Testosterone, Insulin, HOMA-IR and Glucose in C+ve group compared to C-ve group while the treated groups (G1 and G2) showed significant decrease in a parameters compared with C+ve except Insulin levels in G2 which show non-significant difference compared to C-ve, C+ve and G1.

Keywords: PCOS, AMH, HOMA-IR, R. ribes

AbdulWahed AMH, Al-obaidi SMR, Al-samarrai A-MHM (2018) Effect of hot aqueous extract of Rheum ribes roots on some hormonal and biochemical parameters in induced Polycystic Ovary Syndrome in local female rabbits. Eurasia J Biosci 12: 419-423.

INTRODUCTION Rheum ribes one of the most important species in polygonaceae family which contains perennial herbs Polycystic Ovary Syndrome (PCOS) one of the most and its distributes in temperate regions mainly in west common endocrine disorders in women at reproductive Asia countries (Turkey, Iraq, Iran, Lebanon, Syria). Local age (Kavitha et al. 2016). It effects in (6-15)% of women name is Rewas (Sindhu et al. 2016). Phytochemical according to studied population and used criteria studies for this plant shows presense many of active (Smaism, 2016). PCOS defined first time by stein and compounds such as emodin, aloeemodin, physcion, leventhal (1935) when they describe several women had chrysophanol, rhein, rutin (Naemi et al. 2014), it contain oligoamenorea associated with presence of small also vitamin A, B1, B2, C. ovarian cysts in both sides of ovary (Stein and Leventhal R. ribes used to treatment diabetes milletus typeII 1935). It caused by both of genetic and environmental and obesity (Hamzeh et al. 2014). It have antioxidant factors (Yang et al. 2018). PCOS features include properties (Yildirim et al. 2015). This study aimed to complication associated with ovary dysfunction and investigate effect of Rheum ribes on some hormones reproductive disturbances (Balen 2017). levels in induced PCOS in local female rabbits. Metformin is insulin sensitizer uses in inhibition Plant Collection ovarian androgen production and improve ovulation in Rheum ribes roots was collected from local markets PCOS patients (Soyman et al. 2011), contributes in of Kirkuk city, this roots and powdered with electric reduce hepatic glucose production, fatty acids synthesis pestle and kept in closed box until uses. in liver, skeletal muscles and adipose tissues (Pedersen et al. 2018). Plants that have pharmacological properties and medicinal advantages describes as medicinal plants

(Hassan, 2012). Medicinal plants is importance source of antioxidants which used to treatments diseases Received: January 2018 around the world (Rafieian-Kopaie and Badaran 2013, Accepted: August 2018 Kubentayev et al. 2018). Printed: November 2018

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EurAsian Journal of BioSciences 12: 419-423 (2018) AbdulWahed et al.

Table 1. Effect of hot aqueous extract and metformin on some hormoneal and biochemical parameters Insulin Testosterone AMH Glucose HOMA-IR T mIU/ml ng/ml pg/ml 79.2 ± 12.206 1.539±0.375 7.896±1.700 1.912±0.700 4.850±1.216 C-ve c c c bc b 104.706± 17.165 2.753±0.745 10.650±2.193 4.003±0.809 7.057±1.885 C+ve a a a a a 77.275 ±12.963 1.600±0.356 8.380±1.150 2.167±0.599 4.551±0.615 G1 c c bc b b 87.273 ±20.301 1.975±0.499 9.180±1.808 1.548±0.333 5.237±2.122 G2 b b abc c b

Preparation the Hot Aqueous Extract Hormonal Analysis 50g weight of powder roots soaked in 1000ml hot The concentration of serum anti-mullerian, distill water and left 24hours, and filtered with filter testosterone, insulin hormones were measured via papers. The extract dried in electric oven under 40oC. enzyme linked immuno sorbent assay (ELISA) with Metformin Drug using of the commercial kits (ELISA kit, Human, Metformin drug active substrate obtained from SDI Dimeditic, Germany) and the procedure was followed as (The State Company for Drugs Industry & Medical given in the kit catalog. Appliances - Samarra), which was white color powder. Biochemical Analysis Animals Fasting blood sugar concentration quantified by 40 adult female local rabbits weighting (1000-1600) following the procedure that given with kit (spinreact, g and aged (4-6) months selected for the study. The Spain), insulin resistance estimated by using animals housed in wood cages (5 rabbits/cage), homeostatic model assessment of insulin resistance maintained under 12/12h dark/light and fed with which equil: standard laboratory food pallet and tap water. HOMA (IR) = Fasting Insulin (µIU/ml) X Fasting blood sugar (mg/dl)/405 EXPERIMENTAL DESIGN HOMA (IR): Homeostasis model assessment (Bhosle et al. 2016, Ghaffari et al. 2016, Song et al. Determination of Active Dose 2017). Active dose for extracts was determined by using 12 animals, divided randomly for 4 groups. Every group Statistical Analysis All values expressed as mean ±SD data analyzed contain 3 animals. The groups injected subcutaneous done by using analysis of variance (ANOVA). with extracts at control groups injected subcutaneous with distill water. After 3h, the blood sample collected by cardiac puncture and estimate glucose level the dose RESULTS AND DISCUSSION that more hypoglycemic was active dose (Al-Esawy The results show in Table 1 significant increase in 2013). levels of AMH, Insulin, Testosterone and HOMA-IR is Induction of PCOS C+ve group compared to C-ve and treated group show PCOS induced in rabbits by inject (100mg/Kg) significant decrease compared to C+ve. Theory that intraperitoneal daily for 4days consecutive and left 3 explain increase levels of AMH is due to increase days for the developing of syndrome (Bhuvaneshwari et number of small ovarian follicles (Das et al., 2008). Many al. 2015). studies reported there is positive correlation between follicles number and AMH levels (Chen et al. 2008, Animals Divided Piltonen et al. 2005). In this experiment female rabbits randomly divided AMH severity correlate with hyperandrogenism and into four groups: ovulation in PCOS (De leo et al., 2016). This increase in Negative control (C-ve) that receive no thing only AMH levels in PCOS group agreement with Cassar et al. water and food, Positive control (C+ve) injected with (2014) and Dewailly et al. (2016). testosterone propionate (TP), group1 (G1) treated with TP injection causes increase testosterone levels in hot aqueous extract of Rheum ribes roots orally C+ve group which lead to disorders in GnRH feedback (300mg/Kg) and group2 (G2) treated with metformin in PCOS (Chaudhari and Nampothiri 2017). Further TP drug orally (20mg/Kg) period treated was 30days. causes convert testosterone E2 which mean difficult to Blood Sampling recognize it through androgen mediated mechanism After 3 days of treated, blood collected by cardiac (Barraclough 1961), also insulin levels and HOMA-IR puncture and centrifuged at 3000rpm for 10min. Serum increases in C+ve group compared to C-ve, this may be 0 was separated and stored at 20 C until analyzed for due to impair insulin synthesis compared to secretion hormonal and biochemical analysis. PCOS, IR caused by reduction in hepatic clearance and impair pancreatic sensitivity to insulin which

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EurAsian Journal of BioSciences 12: 419-423 (2018) AbdulWahed et al. compensatory cause hyperinsulinemia (Waterworth et al. 2007), which made this plant effective in treatment of al. 1997), IR lead to dyslipidemia and PCOS (Ebrahimi- PCOS. Mamaghani et al. 2015). Glucose levels shows increase G2 shows decrease in AMH, Insulin, Testosterone, in C+ve group compared to C-ve group. This agreement HOMA-IR and Glucose levels, this decrease may be with Maleedhu et al. (2014) who reported that increase through effect of Metformin on glucose levels and insulin glucose levels caused by imbalance between insulin (Adham and Naqishbandi 2015). Many studies reported sensitivity and production when produces resistance that metformin can contribute in improve insulin and finally increase glucose levels in blood (Tarkun et al. resistance through reduction of glucose absorption, 2004). suppress gluconeogenesis and hepatic glucose G1 shows decrease in levels of AMH, Testosterone, production (Bargiota and Diamanti-Kandarakis 2012). Insulin, HOMA-IR and glucose, this may be through Metformin use in PCOS patient to treatment or effects of plant extract on insulin and glucose levels due improvement insulin sensitivity (Shah and Patel 2014). to its contents of polyphenols and active compound Kurzthaler et al. (2014) reported that metformin (emodin, Aloemodin, quercetin, rutin, gallic acid) which inhibit androgen production in ovarian that theca cells. have activity to reduce glucose levels, androgen levels, Also Fiza et al. (2013) suggested that metformin can insulin and HOMA-IR (Gothai et al. 2016, Jahan et al. reduce AMH level in PCOS. 2018, Malaguti et al. 2008, Shah and Patel 2016). Metformin can increase sex hormone binding Further R. ribes have antioxidant properties (Ozturk et globulin (SHBG) in liver so causes reduce in testosterone and AMH levels (Li et al. 2014).

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