桂枝茯苓丸对痰湿型多囊卵巢综合征模型大鼠的改善作用及机制研究
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篇名: 桂枝茯苓丸对痰湿型多囊卵巢综合征模型大鼠的改善作用及机制研究
TITLE: Study on Improvement Effects and Mechanism of Guizhi Fuling Pills on Phlegm-dampness Type PCOS Model Rats
摘要: 目的:研究桂枝茯苓丸对痰湿型多囊卵巢综合征(PCOS)模型大鼠的改善作用及可能机制。方法:将50只动情周期规律的雌性SD大鼠随机分为空白对照组(n=10)和造模组(n=40)。造模组大鼠采用灌胃来曲唑[1mg/(kg·d),连续21d]的方法建立痰湿型PCOS模型。将造模成功的40只大鼠再随机分为模型组、二甲双胍组[阳性对照,0.135g/(kg·d)]和桂枝茯苓丸低、高剂量组[4.49、17.96g/(kg·d)],每组10只。各药物组大鼠均灌胃给药,每天1次,连续给药30d;空白对照组和模型组大鼠灌胃等体积生理盐水。末次给药结束后,采用口服葡萄糖耐量实验检测大鼠灌胃葡萄糖溶液0、30、60、120min后的血糖水平;采用酶联免疫吸附法检测大鼠血清中促卵泡生成激素(FSH)、促黄体生成素(LH)、雌二醇(E2)、睾酮(T)水平;称定大鼠体质量和子宫、双侧卵巢、性腺旁脂肪的质量;采用苏木精-伊红染色法观察大鼠卵巢组织的形态学变化;分别采用实时荧光定量-聚合酶链式反应法和West-ernblotting法检测大鼠卵巢组织中芳香化酶(CYP19a1)和子宫组织中葡萄糖转运蛋白4(Glut4)的mRNA及蛋白表达水平。结果:与空白对照组比较,模型组大鼠灌胃葡萄糖溶液30、60、120min后的血糖水平以及血清中LH、E2、T水平均显著升高(P<0.05);体质量和子宫、双侧卵巢、性腺旁脂肪的质量均显著增加(P<0.05);卵泡膜细胞体积明显增大、呈圆球形;卵巢组织中CYP19a1和子宫组织中Glut4的mRNA及其蛋白表达水平均显著降低(P<0.05或P<0.01)。与模型组比较,各给药组大鼠上述指标水平均显著改善(P<0.05或P<0.01);桂枝茯苓丸低、高剂量组大鼠卵泡膜细胞层厚度和卵泡膜细胞体积也有一定改善。结论:桂枝茯苓丸能够上调卵巢组织中CYP19a1基因及子宫组织中Glut4基因的表达,从而改善痰湿型PCOS模型大鼠糖代谢异常、减轻胰岛素抵抗、下调血清雌激素水平、减轻卵巢多囊样改变程度,最终起到治疗PCOS的作用。
ABSTRACT: OBJECTIVE:To stu dy the improvem ent effect and mechanism of Guizhi fuling pill (GZFLP) on rats with phlegm-dampness type polycystic ovary syndrome (PCOS).METHODS:Totally fifty female SD rats with regular estrous cycle were randomly divided into blank control group (n=10) and model group (n=40). The model group received letrozole intragastrically [1 mg/(kg·d),for 21 d ] to induce phlegm-damp PCOS rat model. Forty model rats were randomly divided into model group ,metformin group [positive control ,0.135 g/(kg·d)] and GZFLP low-dose ,high-dose groups [4.49,17.96 g/(kg· d)],with 10 rats in each group. Each medicine group was given intragastric administration ,once a day ,for 30 consecutive days. Blank control and model group were given intragastric administration of equal volume normal saline.After last medication ,oral glucose tolerance test (OGTT)was used to detect the level of blood glucose in rats at 0,30,60 and 120 min. ELISA assay was used to detect the levels of follicle-stimulating hormone (FSH),luteinizing hormone (LH),estradiol(E2)and testosterone (T); body weight ,the weight of uterus ,bilateral ovaries and paragonadal fat were weighed. HE staining was used to observe morphological changes of ovarian tissue sections. RT-PCR method and Western blotting assay were used to detect mRNA and protein expression of CYP 19a1 in ovary and Glut 4 in uterine. RESULTS :Compared with blank control group ,the levels of blood glucose 30,60,120 min after intragastric administration of glucose solution as well as serum levels of LH ,E2 and T were increased significantly in model group (P<0.05);the body weight ,the weight of uterus ,bilateral ovaries and paragonadal fat were increased significantly (P<0.05);the volume of thecal cells increased significantly and showed spherical shape ;the mRNA and protein expression of CYP 19a1 in ovarian tissue and Glut 4 in uterine tissue were decreased significantly (P<0.05 or P<0.01). Compared with model group ,above indexes of administration groups were improved significantly (P<0.05 or P<0.01);the thickness of theca cell layer and the volume of theca cells in GZFLP low-dose and high-dose groups were also improved. CONCLUSIONS:GZFLP can up-regulate the expression of CYP19a1 in ovary and Glut4 in uterus ,so as to improve the abnormal glucose metabolism in phlegm-dampness type PCOS model rats ,reduce insulin resistance ,lower serum estrogen levels ,and reduce the degree of polycystic changes in ovary ,thereby playing a role in the treatment of PCOS.
期刊: 2021年第32卷第01期
作者: 余璟玮,钟晓莹,曾蕾,郑泳霞
AUTHORS: YU Jingwei ,ZHONG Xiaoying ,ZENG Lei,ZHENG Yongxia
关键字: 桂枝茯苓丸;痰湿型多囊卵巢综合征;胰岛素抵抗;芳香化酶;葡萄糖转运蛋白4
KEYWORDS: Guizhi fuling pills ;Phlegm-dampness type polycystic ovarian syndrome ;Insulin resistance ;Aromatase;Glucose
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