高氏盆炎方三号对慢性盆腔炎大鼠炎症介质和生殖功能的影响

Effect of Gao's Prescription No.3 on Inflammatory Factors and Reproductive Function in Rats with Chronic Pelvic Inflammatory Disease

目的:观察高氏盆炎方三号对慢性盆腔炎大鼠炎症介质和生殖功能的影响并探讨其可能的作用机制。方法:将119只SD大鼠分为药效学观察组(药效组)和生殖功能观察组(生殖组),采用机械损伤联合化学烧伤方法制备慢性盆腔炎大鼠模型(10 d),并按随机数字表法分为模型组、蒲苓组、人胎盘组织液组(简胎盘组)、高氏盆炎方三号高、中、低剂量组(中药高、中、低剂量组),另设正常动物为空白组,药效组,每组10只,生殖组每组7只,给药20 d。药效组处死大鼠,酶联免疫吸附试验法检测粒细胞-巨噬细胞集落刺激因子(GM-CSF)、白细胞介素(IL)-4的水平;光镜下观察各组大鼠子宫病理学变化;免疫组化法和RT-PCR法分别检测核因子κB蛋白和mRNA的表达。生殖组以7﹕2比例进行雌雄合笼,观察60 d内大鼠妊娠率、产仔数等变化。结果:光镜下模型组大鼠子宫组织呈现明显炎细胞浸润,内膜充血水肿,管腔壁增厚,组织结构紊乱,腺体紊乱萎缩变形等病理改变。与模型组比较,高氏盆炎方三号高、中、低剂量组大鼠血清GM-CSF水平均显著降低,IL-4水平升高,大鼠子宫组织核因子κB蛋白和基因表达均明显降低(P<0.05);生殖组中模型组妊娠率为0,中药高、中、低剂量组总妊娠率较高,且大鼠受孕率、产仔数、平均产仔数及子鼠存活率都较高,中药高剂量组受孕率与空白组比较差异无统计学意义(P>0.05)。结论:高氏盆炎方三号对慢性盆腔炎大鼠及炎性不孕大鼠有良好的治疗作用,其作用机制可能是通过促进抗炎细胞因子IL-4分泌水平,抑制核因子κB的激活,进而降低GM-CSF释放,减轻盆腔炎炎症反应,改善大鼠的生殖功能。

Objective:To investigate the effect of Gao's Prescription No.3 on inflammatory factors and reproductive function in rats with chronic pelvic inflammatory disease(CPID)and explore its possible mechanism.Methods:A total of 119 SD rats were randomly divided into two major groups,namely the pharmacodynamics experimental groups(referred to as the pharmacodynamic groups)and the reproductive function experimental groups(referred to as the reproductive groups).The rat model of CPID was prepared by mechanical injury combined with chemical burn for 10 days.The rats with CPID were randomly divided into a model group,a Puling group,a placenta group,and three high,medium,and low-dose Gao's Prescription No.3 groups.The normal rats were divided into the control group.There were 10 rats in each pharmacodynamic group and 7 rats in each reproductive group.The administration lasted for 20 days.Rats in the pharmacodynamic groups were sacrificed,and the levels of granulocyte-macrophage colony-stimulating factor(GM-CSF)and interleukin-4(IL-4)were determined by enzyme-linked immunosorbent assay(ELISA).The pathological changes in the uterus of rats in each group were observed under a light microscope.Immunohistochemical assay and real-time quantitative polymerase chain reaction(RT-PCR)were used to determine the protein and mRNA expressions of nuclear factor kappa B(NF-κB),respectively.In the reproductive groups,the male and female rats were caged at a 7∶2 ratio,and the changes in pregnancy rate and litter size in rats within 60 days were observed.Results:Under the light microscope,the model group showed obvious inflammatory cell infiltration,intimal hyperemia and edema,thickening of the lumen wall,tissue structure disorder,gland disorder and atrophy,and other pathological changes in the uterine tissue of rats.As compared with the model group,the GM-CSF levels in the high,medium,and low-dose Gao's Prescription No.3 groups were decreased,IL-4 levels were increased,and the protein and mRNA expressions of NF-κB were reduced(P<0.05).In the reproductive groups,the pregnancy rate of the model group was 0,and the total pregnancy rate of the high,medium,and low-dose Gao's Prescription No.3 groups was high,with high litter size,average litter size,and survival rate.There was no statistical difference in the pregnancy rates between the high-dose Gao's Prescription No.3 group and the control group(P>0.05).Conclusion:Gao's Prescription No.3 has a good therapeutic effect on rats with CPID and infertility.The mechanism may be related to the promotion of the secretion level of anti-inflammatory cytokine IL-4 and the inhibition of activation of NF-κB,thus reducing the release of GM-CSF,alleviating the response of PID,and improving the reproductive function of rats.