【作者】 韩旭；

【导师】 马宝璋；

【作者基本信息】 黑龙江中医药大学 ， 中医妇科学， 2009， 博士

【摘要】 目的：本实验通过对炎克宁V号方作用于慢性盆腔炎模型大鼠其免疫调节、凋亡调控、信号传导等关键产物的观察分析，探讨CPID的发病机制及炎克宁V号方治疗的作用机理，为炎克宁V号方治疗CPID的临床应用和推广提供理论基础和实验依据。方法：以Wistar雌性大鼠为受试对象，用混合菌(大肠杆菌、乙型溶血性链球菌和金黄色葡萄球菌)接种法建立慢性盆腔炎大鼠模型。将69只大鼠随机分为正常组、假手术组、模型组、炎克宁V号方高、中、低剂量组和中药对照组。分别应用光镜和电镜方法对各组大鼠子宫组织进行形态学观察；应用免疫组织化学反应法检测各组大鼠子宫组织中INF-γ和Caspase-3的表达；应用酶联免疫吸附测定(ELISA)法检测各组大鼠血清中IL-2和IL-6含量；应用逆转录聚合酶链式反应(RT-PCR)法检测各组大鼠子宫组织MMP-2 mRNA，Fas／FasL mRNA和p53 mRNA的表达，硝酸还原法检测血清中NO含量及化学比色法测定血清NOS活性。结果：慢性盆腔炎模型组大鼠血清中IL-2、IL-6、NO的含量、子宫组织中INF-γ的表达、NOS的活性均明显高于正常组(P＜0．05)，子宫组织中Caspase-3，MMP-2mRNA，Fas／FasLmRNA和p53mRNA的表达均明显低于正常组(P＜0．05)。炎克宁V号方明显降低了慢性盆腔炎模型大鼠血清中IL-2、IL-6、NO的含量及NOS的活性(P＜0．05)，明显减弱了模型大鼠子宫组织中INF-γ的表达(P＜0．05)，同时增强了模型大鼠子宫组织中Caspase-3，MMP-2 mRNA，Fas／FasL mRNA和p53 mRNA的表达(P＜0．05)。其中，炎克宁V号中剂量组的作用均明显优于中药对照组(P＜0．05)。结论：炎克宁V号方通过降低慢性盆腔炎模型大鼠血清中IL-2，IL-6含量，使它们接近正常水平，恢复免疫平衡，从而控制慢性盆腔炎的病理发展；通过增强模型大鼠子宫组织中MMP-2 mRNA的表达，降解细胞外基质，从而减轻慢性盆腔炎的纤维组织增生；通过增加模型大鼠子宫组织中Caspase-3，Fas／FasL mRNA和p5 3 mRNA的表达，诱导炎细胞凋亡，抑制炎细胞生长，从而减轻炎症反应，有利于慢性盆腔炎症的消除。炎克宁V号能明显降低NO含量和NOS的活性，减轻组织细胞的损伤，从而抑制炎症细胞的形成，达到CPID的治疗作用。综上，炎克宁V号方是治疗慢性盆腔炎的有效方剂。更多还原

【Abstract】 Objective:To explore the effect of a defined formula of Chinesemedicinal herbs-Yankening V (also known as the pelvic inflammationrelieving formula V)and the mechanism of Yankening V in treating CPIDon the model rats with chronic pelvic inflammatory disease (CPID)byobserving the roles that it may play in the processes of immunoregulation,the control of cell apoposis and signal transduction,which could provideevidence to the clinical application of YankeNing V.Methods:The models of rats with CPID (Wistar female rats)weresuccessfully made by infecting three various bacteria into the rats.Sixty-seven female Wistar rats were randomly classified into the followingdifferent groups:the normal group,the simulative model group,the modelgroup,the high dose of YankeningV group,the medium dose of YankeningV group,the low dose of YankeNing V group and the control group withanother kind of Chinese medicinal formula.The light microscope andelectron microscope were used in the morphologic research and the methodof immuno-histochemistry was used to detect the expression of INF-γandCaspase-3 in the uterine tissues of the model rats with CPID.The method ofenzyme linkedimmunosorbent assay (ELISA)was used to detect the contentof IL-2 and IL-6 in the serum of the model rats.The method of reversetranscription follow by PCR (RT-PCR)was used to detect the expression ofINF-γmRNA,MMP-2mRNA,Fas/FasLmRNA and p53mRNA in the uterinetissues of the model rats with CPID.The content of NO and NOS in serumwas respectively detected with the method of nitrate reductase and chemicalcolorimetry.Results:The contents ofIL-2,IL-6 and NO in the serum of the modelrats were significantly higher than those of the normal rats (P＜0.05),sowere the activity of NOS and the expressions of INF-γin the uterine tissuesof the model rats with CPID (P＜0.05).The expressions of Caspase-3,Fas/FasLmRNA,p53mRNA and MMP-2mRNA in the uterine tissues of the model rats were significantly weaker than those of the normal rats (P＜0.05).Yankening V has significantly decreased the contents ofIL-2,IL-6 and NOin the serum of the model rats,the activity of NOS and the expressions ofINF-γin the uterine tissues of the model rats with CPID(P＜0.05).It has alsosignificantly increased the expressions of Caspase-3,MMP-2mRNA,Fas/FasLmRNA and p53mRNA in the uterine tissues of the model rats(P＜0.05).The medium dose of Yankening V group functioned significantlybetter than the control group with another kind of Chinese medicine(P＜0.05)Conclusions:It was through decreasing the contents of IL-2 and IL-6 inthe serum of the model rats and the expressions of INF-γin the uterinetissues of the model rats with CPID that Yankening V play the roles ofrestore the immune disequilibrium,thus relieving the pelvic inflammation.Itwas through increasing the expressions of MMP-2 mRNA in the uterinetissues of the model rats that Yankening V degrades extracellular matrix andinhabits the fibroplasia and then facilitate to remove CPID.It was throughincreasing the expressions of Caspase-3,Fas/FasL mRNA and p53 mRNA inthe uterine tissues of the model rats that Yankening V plays the roles ofinducing the programmed inflammation cell death,thus relieving andremoving CPID.It can be concluded that the effects of degrading the lever ofNO and the activity of NOS by Yankening V was through decreasing theinjury of the cell to treat CPID.Yankening V is a kind of effective definedformulas of Chinese medicinal herbs in treating CPID.更多还原