癸源煎配方颗粒对卵巢储备功能下降模型小鼠的改善作用及机制研究-中国药房

癸源煎配方颗粒对卵巢储备功能下降模型小鼠的改善作用及机制研究

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篇名：	癸源煎配方颗粒对卵巢储备功能下降模型小鼠的改善作用及机制研究
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摘要：	目的：研究癸源煎配方颗粒（GDFG）对卵巢储备功能下降（DOR）模型小鼠的改善作用及机制。方法：将动情周期正常的42只雌性ICR小鼠随机分为对照组、模型组、戊酸雌二醇组（阳性对照，0.15mg/kg）和GDFG低、中、高剂量组（0.75、1.49、2.98g/kg），每组7只。除对照组外，其余各组小鼠均腹腔注射顺铂（3mg/kg）复制DOR模型。造模成功后，给药组小鼠灌胃相应药物，模型组和对照组小鼠灌胃生理盐水，每天1次，连续4周。末次给药后，采用酶联免疫吸附试验测定小鼠血清中抗米勒管激素（AMH）和促卵泡生成素（FSH）水平；采用苏木精-伊红（HE）染色法观察小鼠卵巢组织病理学形态；采用免疫组化法观察小鼠卵巢组织中AMHR受体Ⅱ（AMHRⅡ）、Smad4蛋白的分布情况；采用Westernblot法检测小鼠卵巢组织中AMHRⅡ、Smad4蛋白的表达水平。结果：与对照组比较，模型组小鼠血清中AMH水平和卵巢组织中AMHRⅡ、Smad4蛋白表达水平均显著降低（P＜0.01），血清中FSH水平显著升高（P＜0.01）；卵泡组织中可见卵泡皱缩、卵泡核丢失、卵巢间质纤维化、黄体疏松；AMHRⅡ、Smad4蛋白主要分布在卵泡膜上和卵巢间质中。与模型组比较，GDFG各剂量组小鼠血清中AMH水平和卵巢组织中AMHRⅡ、Smad4蛋白表达水平均显著升高（P＜0.01），血清中FSH水平均显著降低（P＜0.05或P＜0.01）；卵巢组织中可见各级卵泡、卵泡形态改善，未见明显核丢失和卵丘形成；AMHRⅡ、Smad4蛋白主要分布在卵泡核（GDFG高剂量组除外）及卵巢颗粒细胞膜上（GDFG中剂量组以分布在窦卵泡为主），成熟卵泡核周围或黄体中有少许分布。结论：GDFG可改善DOR模型小鼠的卵巢功能，其机制可能与升高血清中AMH水平和卵巢组织中AMHRⅡ、Smad4蛋白表达水平，改善AMHRⅡ、Smad4蛋白在卵巢颗粒细胞膜和卵泡核内的分布以及降低血清中FSH水平有关。
ABSTRACT：	OBJECTIVE：To study the improvement eff ects and its mech anism of Guiyuan decoction formula granules （GDFG） on model mice with decreased ovarian reserve （DOR）. METHODS ：Totally 42 female ICR mice whith with normal estrous cycle were randomly divided into control group ，model group ，estradiol valerate group （positive control ，0.15 mg/kg）and GDFG low-dose，medium-dose and high-dose groups （0.75，1.49，2.98 g/kg），with 7 mice in each group. Except for control group ，other groups were given cisplatin （3 mg/kg）intraperitoneally to establish DOR model. After modeling ，administration groups were given relevant medicine intragastrically；model group and control group were given normal saline intragastrically ，once a day ，for consecutive 4 weeks. After last administration ，ELISA assay was used to measure the serum levels of anti-Müllerian hormone （AMH）and follicle-stimulating hormone （FSH）in mice. Histopathological morphology of ovarian was observed by HE staining. Protein distribution of AMH receptor Ⅱ（AMHRⅡ）and Smad 4 in ovarian tissue were observed by immunohistochemistry. Protein expression of AMHR Ⅱ and Smad 4 were detected by Western blot assay. RESULTS ：Compared with control group ，theserum level of AMH ，the expression of AMHR Ⅱ and Smad 4 protein in ovarian tissue in model group were significantly decreased （P＜0.01），while the FSH level in serum was significantly increased （P＜0.01）；follicles were crumpled and lost nucleus ，ovarian interstitial were fibrosis ，luteum were loose ； AMHRⅡ and Smad 4 protein in ovarian tissue were mainly distributed in the follicle membrane and ovarian interstitial. Compared with model group ，the serum level of AMH ，the expression of AMHR Ⅱ and Smad 4 protein in ovarian tissue was increased significantly in GDFG groups （P＜0.01），while the serum level of FSH was decreased significantly （P＜0.05 or P＜0.01）；in ovarian tissue ，follicles at all levels could be found and follicle morphology was improved ，and no obvious nuclear loss and cumulus formation were found ；AMHRⅡ and Smad 4 protein were mainly distributed in the follicular nucleus （except for GDFG high-dose group) and the granular cell membrane （mainly distributed in the sinus follicles of GDFG medium-dose group ）；they were slightly distributed around the mature follicular nucleus or in corpus luteum. CONCLUSIONS ：GDFG can improve ovarian function of DOR model mice. The mechanism may be related with promoting serum level of AMH ，protein expression of AMHR Ⅱ and Smad 4，improving the distribution of AMHR Ⅱ and Smad 4 protein in ovarian granulosa cell membrane and follicular nucleus ， reducing FSH levels.
期刊：	2021年第32卷第09期
作者：	王月娇，孙兆贵，徐莲薇，郁琳，张瑜，刘小菲，李盛楠，丛超，赵莉
AUTHORS：
关键字：	卵巢储备功能下降；癸源煎配方颗粒；抗米勒管激素；促卵泡生成素；抗米勒管激素Ⅱ受体；Smad4；小鼠
KEYWORDS：	Decreased ovarian reserve ；Guiyuan decoction formula granules ；Anti-Müllerian hormone ；Follicle stimulating
阅读数：	285 次
本月下载数：	5 次

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