Balasubramanian Arul, Pachiappan Sudhakar, Mohan Surendiran, Adhikesavan Harikrishnan, Karuppasamy Indira, Ramalingam Kothai
Department of Pharmacology, Vinayaka Mission's College of Pharmacy,Vinayaka Mission's Research Foundation (Deemed to be University), Salem, 636308, Tamil Nadu, India.
Department of Pharmacology, Swamy Vivekanandha College of Pharmacy, Elayampalayam, Tiruchengode, Tamil Nadu, India.
Heliyon. 2023 Apr 21;9(5):e15488. doi: 10.1016/j.heliyon.2023.e15488. eCollection 2023 May.
This study aimed to develop an effective alternative medicine with multi potential herbs against polycystic ovarian syndrome (PCOS) in rats induced by letrozole treatment.
Polyherbal syrup was prepared with a combination of bark leaves aerial parts stem bark seeds, and roots ethanolic extract. cell viability study, adenosine monophosphate-activated protein kinase (AMPK), and glucose transporter 4 (GLUT4) gene expression assay were carried out on the Chinese Hamster Ovarian (CHO) cell line. For the PCOS induction letrozole (1 mg/kg ) was given for 21 consecutive days. The PCOS induction was confirmed by measuring estrus irregularity, insulin resistance by oral glucose tolerance test (OGTT), and hyperandrogenism by measuring serum total testosterone level 21 days after completion of letrozole treatment. After induction of PCOS, metformin (155 mg/kg ), and polyherbal syrup (100 mg/kg, 200 mg/kg, and 400 mg/kg ) were administered for further 28 days. The treatment efficacy was measured by measuring serum lipid profile, fasting insulin level, sex hormones level, ovarian steroidogenic enzymes, ovarian tissue insulin receptor, AMPK, and GLUT4 protein expression levels, and histomorphological studies. The post-treatment effect was confirmed by reproductive performance studies.
Letrozole-induced PCOS rats showed significant estrus irregularity, abnormal sex hormones levels, and hyperandrogenism indicated by showing increased free androgenic index and decreased sex hormones binding globulin (SHBG) level. The insulin resistance in PCOS rats was indicated by increased fasting glucose levels with impaired glucose clearance in the OGT test. Homeostasis Model Assessment Index of Insulin Resistance (HOMA-IR) increased level, also decreases INSR, GLUT4, and AMPK mRNA expression in ovarian cells confirming the insulin resistance in PCOS rats. Ovarian histology in PCOS rats also showed many follicular cysts, atretic follicles, and the absence of corpus luteum. The administration of polyherbal syrup, in a dose-dependent manner, effectively restored these alterations. The treatment of polyherbal formulation 400 mg/kg possesses highly significant efficacy over the treatment of metformin in PCOS rats. It mainly acts by reducing peripheral and ovarian hyperandrogenism and improves insulin sensitivity via activating the insulin receptor and AMP-activated kinase-mediated transcription and translation of GLUT4 from the cytoplasm to the ovarian membrane improves glucose uptake and promotes the follicular development and ovulation. The higher fertility rate, delivery index, and survival of delivered pups confirm the broader and superior efficacy of PCOS. These beneficial actions are mainly attributable to the formulation's inclusion of the key secondary metabolites flavonoids and phytosterols. In conclusion, the prepared polyherbal syrup was found to be the safest and most effective alternative medicine for both endocrinal and metabolic complications of PCOS women
本研究旨在开发一种由多种具有潜在功效的草药制成的有效替代药物，用于治疗来曲唑诱导的大鼠多囊卵巢综合征（PCOS）。
用树皮叶、地上部分、茎皮、种子和根的乙醇提取物制成多草药糖浆。在中国仓鼠卵巢（CHO）细胞系上进行细胞活力研究、单磷酸腺苷激活蛋白激酶（AMPK）和葡萄糖转运蛋白4（GLUT4）基因表达测定。为诱导PCOS，连续21天给予来曲唑（1mg/kg）。通过测量发情周期不规律、口服葡萄糖耐量试验（OGTT）评估胰岛素抵抗以及在来曲唑治疗完成21天后测量血清总睾酮水平评估高雄激素血症，以确认PCOS的诱导情况。诱导PCOS后，再给予二甲双胍（155mg/kg）和多草药糖浆（100mg/kg、200mg/kg和400mg/kg）28天。通过测量血清脂质谱、空腹胰岛素水平、性激素水平、卵巢类固醇生成酶、卵巢组织胰岛素受体、AMPK和GLUT4蛋白表达水平以及组织形态学研究来评估治疗效果。通过生殖性能研究确认治疗后的效果。
来曲唑诱导的PCOS大鼠表现出明显的发情周期不规律、性激素水平异常和高雄激素血症，表现为游离雄激素指数升高和性激素结合球蛋白（SHBG）水平降低。PCOS大鼠的胰岛素抵抗表现为空腹血糖水平升高以及OGT试验中葡萄糖清除受损。胰岛素抵抗的稳态模型评估指数（HOMA-IR）升高，同时卵巢细胞中胰岛素受体（INSR）、GLUT4和AMPK的mRNA表达降低，证实了PCOS大鼠存在胰岛素抵抗。PCOS大鼠的卵巢组织学也显示出许多卵泡囊肿、闭锁卵泡以及黄体缺失。多草药糖浆以剂量依赖的方式有效恢复了这些改变。400mg/kg的多草药制剂治疗PCOS大鼠的疗效显著优于二甲双胍。它主要通过降低外周和卵巢高雄激素血症，并通过激活胰岛素受体和AMP激活激酶介导GLUT4从细胞质到卵巢膜的转录和翻译来提高胰岛素敏感性，从而改善葡萄糖摄取并促进卵泡发育和排卵。较高的生育率、分娩指数和所产幼崽的存活率证实了多草药糖浆对PCOS更广泛和更优的疗效。这些有益作用主要归因于该制剂中含有关键次生代谢产物黄酮类化合物和植物甾醇。总之，所制备的多草药糖浆被发现是治疗PCOS女性内分泌和代谢并发症最安全、最有效的替代药物。
