补肾化痰法调节PCOS卵泡基底膜分子筛功能的研究

Research on using reinforcing kidney and eliminating phlegm method to accommodate the function of molecular sieve in follicle basement membrane of PCOS

目的:从分子生物学水平研究补肾化痰中成药灵术颗粒调控多囊卵巢综合征(PCOS)模型大鼠卵泡基底膜分子筛功能,调节两细胞间信息交换,促进卵泡发育的可能作用靶点,阐明其疗效的分子机制;为灵术颗粒治疗PCOS的临床应用提供科学依据。方法:应用Poresky方法制造PCOS大鼠,采用随机对照实验的方法,分为灵术颗粒组、二甲双胍组、空白对照组及正常对照组,各组连续灌胃14d。测定各组大鼠治疗后卵泡基底膜厚度及颗粒细胞、卵泡内膜细胞数目及卵泡基底膜蛋白(Ⅳ型胶原、层连接蛋白及串珠养蛋白聚糖)表达含量。结果:4组大鼠治疗后卵泡基底膜厚度均数比较,空白对照组基底膜厚度显著高于其他3组,而灵术颗粒组与二甲双胍组基底膜厚度接近正常对照组。4组间大鼠卵泡基底膜Ⅳ型胶原、连接蛋白及串珠养蛋白聚糖的表达有显著差异。Ⅳ型胶原的比较,空白对照组表达最多,灵术颗粒组次之,正常对照组再次之,二甲双胍组最少。而层连接蛋白和串珠养蛋白聚糖的比较,以空白对照组表达最多,灵术颗粒组与二甲双胍组效果无差别,正常对照组层连接蛋白表达最少。结论:补肾化痰中成药灵术颗粒和二甲双胍均可通过减少PCOS卵泡基底膜蛋白表达,改变卵泡基底膜厚度,改善卵泡基底膜分子筛功能,调节两细胞间信息交换,促进卵巢排卵;灵术颗粒和二甲双胍效果相当。

Objective：Study how the reinforcing kidney and eliminating phlegm method regulating and controlling the function of molecular sieve in follicle basement membrane in polycystic ovarian syndrome（PCOS） model rats from the level of molecular biology.Study how it adjusted information exchange between two cells,maintain the function balance between ‘LHtheca cells axes＇ and ‘FSH-granular cell axes＇,and promote the possible targets of follicular development.Clarify the molecular mechanisms of its efficacy.Provide a scientific basis for using reinforcing kidney and eliminating phlegm method in clinical treatment of PCOS.Methods： This study is basis of Poresky Methods to make the formers of PCO rats.The rats were divided into four groups by randomization and comparison,study group,control group,blank control group and normal control group.Each group of rats was received intragastric administration for 14 days.Determined each group of rats with the thickness of follicle basement membrane and the quantity of granular cells and theca cells,and the content of follicular basement membrane proteins（collagen Ⅳ,Laminin and Perlecan）.Result：The comparison of the mean thickness of follicle basement membrane of 4 groups of rats after treatment showed that the thickness of blank control group was significantly higher than the other three groups,while the thickness of basement membrane of the study group and control group were close to the normal control group.The blank control group had the least number of granule cells.It showed that there was significant difference between the 4 Groups in measurement of follicular basement membrane of collagen Ⅳ,Laminin and Perlecan.Comparison of the thickness of collagen Ⅳ showed that the blank control group is the most of the expression of collagen Ⅳ,the second is the study group,the third is the normal control group,the last is the control group.The comparison of the expression of Laminin and Perlecan showed that the blank control group is the most of the expression of both,the second is the study group and the control group; the last is the normal control group.Conclusion.The Chinese medicine Ling-Zhu Granule,which based on the reinforcing kidney and eliminating phlegm method to treat PCOS could reduce the expression of follicle basement membrane protein,alter the thickness of follicle basement membrane,improve the function of molecular sieve in follicle basement membrane,adjusted information exchange between two cells,maintain the function balance between ‘LH-theca cells axes＇ and ‘FSH-granular cell axes＇,promote ovulation as well as Metformin.