Zihuai recipe alleviates cyclophosphamide-induced diminished ovarian reserve via suppressing PI3K/AKT-mediated apoptosis

Autor: Shaohu Zhou, Qi Chen, Zhiwei Weng, Jiaming Feng, Zhidan Liu, Truong Nam Nguyen, Weiping Liu
Rok vydání: 2020
Předmět:
endocrine system
medicine.medical_specialty
Hypothalamo-Hypophyseal System
Hormone Replacement Therapy
medicine.medical_treatment
Intraperitoneal injection
Hypothalamic–pituitary–gonadal axis
Apoptosis
Rats
Sprague-Dawley
03 medical and health sciences
Phosphatidylinositol 3-Kinases
0302 clinical medicine
Ovarian Follicle
Internal medicine
Drug Discovery
medicine
Medroxyprogesterone acetate
Animals
Ovarian reserve
Ovarian Reserve
Antineoplastic Agents
Alkylating
Cyclophosphamide
030304 developmental biology
Pharmacology
0303 health sciences
Granulosa Cells
Dose-Response Relationship
Drug
Estradiol
business.industry
Estradiol valerate
Antral follicle
Rats
Endocrinology
030220 oncology & carcinogenesis
Female
Luteinizing hormone
business
Proto-Oncogene Proteins c-akt
medicine.drug
Hormone
Drugs
Chinese Herbal
Zdroj: Journal of ethnopharmacology. 277
ISSN: 1872-7573
Popis: Ethnopharmacological relevance Zihuai recipe (ZHR), a Chinese herbal prescription, is widely used for the clinical treatment of Diminished ovarian reserve (DOR) infertility. However, little is known regarding its underlying mechanisms of DOR treatment. Aim of the study This study aimed to investigate the beneficial effects of ZHR on the treatment of DOR and to reveal the underlying mechanisms. Materials and methods Sixty 8-week-old Sprague–Dawley female rats were randomly divided into the following six groups (n=10 per group): control group, DOR group, low-dose (2.7 g/kg/day) ZHR (L-ZHR) group, medium-dose (5.4 g/kg/day) ZHR (M-ZHR) group, high-dose (10.8 g/kg/day) ZHR (H-ZHR) group, and hormone replacement therapy (HRT) group. The DOR model was established in all the groups, except the control group, by a single intraperitoneal injection of 90 mg/kg cyclophosphamide (CTX). After the induction of the DOR model, rats were weighed and administered either the relevant dose of ZHR or an equal volume of saline solution (in the control and DOR groups). Rats in the HRT group received estradiol valerate tablets (0.16 mg/kg/day) for 4 days， with medroxyprogesterone acetate tablets (0.86 mg/kg/day) added on day 4. After 32 days of treatment, the rats were euthanized and the ovaries were collected for sampling. Ovarian morphology was observed by hematoxylin and eosin staining and the number of follicles was counted under a microscope. The serum levels of anti-Mullerian hormone (AMH), gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) were quantified by ELISA. A TUNEL assay was used to analyze the level of apoptosis of the ovarian cells. The protein expressions of p-PI3K, p-AKT, PI3K, AKT, cleaved caspase-3, BAX, and Bcl-2 were measured by western blotting and immunohistochemistry. Data analysis was performed with SPSS 20.0 software. Results ZHR administration increased the ovarian index and the serum levels of AMH, GnRH, and E2, while it lowered those of FSH and LH. ZHR treatment also increased the numbers of primordial, primary, secondary, and antral follicles, as well as the numbers of corpora lutea, but decreased the number of atretic follicles. Furthermore, ZHR administration decreased the percentage of TUNEL-positive ovarian cells. After treatment with ZHR, the protein expression levels of p-PI3K/PI3K, p-AKT/AKT, cleaved caspase-3 and BAX were decreased, whereas the level of Bcl-2 was increased. Conclusions ZHR improved the ovarian reserve in CTX-induced DOR rats. The mechanisms of ZHR on DOR may be mediated through the regulation of gonadal hormones of the hypothalamic-pituitary-ovarian axis, and the inhibition of PI3K/AKT-mediated apoptosis in granulosa cells.
Databáze: OpenAIRE
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