【作者】 宗惠；

【导师】 刘瑞芬；

【作者基本信息】 山东中医药大学 ， 中医妇科学， 2010， 博士

【摘要】 目的：通过观察慢性盆腔炎模型大鼠血清、子宫组织中炎症细胞因子、粘连相关指标及核因子的变化,探讨活血补肾法治疗慢性盆腔炎(CPID)的抗炎抗粘连的免疫调节机制。方法：以Wistar雌性大鼠为实验对象,采用混合细菌加机械损伤方法,复制慢性盆腔炎动物模型。将65只大鼠随机分为正常对照组、假手术组、模型组、盆腔炎高、中、低剂量组、盆炎净组,酶联免疫吸附测定法(ELISA)检测各组大鼠血清的炎症细胞因子水平,包括白介素-1β(IL-1β)、白介素-8(IL-8)、单核细胞趋化蛋白-1(MCP-1),白介素-10(IL-10)及粘连相关细胞因子转化生长因子-β1 (TGF-β1);大鼠子宫形态学观察后,免疫组织化学法检测各组大鼠子宫组织肿瘤坏死因子-α(TNF-a).细胞间粘附分子-1 (ICAM-1)及核因子κB (NF-κB)、核因子抑制蛋白(IκB)的表达；逆转录-聚合酶链反应法(RT-PCR)检测大鼠子宫组织TNF-amRNA、ICAM-1mRNA的表达。结果：慢性盆腔炎模型大鼠血清前炎症细胞因子IL-1β、IL-8、MCP-1水平较正常组大鼠升高,抗炎症细胞因子IL-10水平较正常对照组大鼠降低,血清中粘连相关细胞因子TGF-β1、子宫组织中的粘连相关指标TNF-α、ICAM-1、TNF-amRNA. ICAM-1mRNA表达及子宫组织NF-κB、IκB表达水平均较正常对照组升高,差异均有极显著性(P<0.01)。药物干预后,各治疗组大鼠血清前炎症细胞因子水平及TGF-β1水平均较模型组下降,抗炎症细胞因子水平则均较模型组升高；各治疗组大鼠子宫组织中TNF-α、ICAM-1、NF-κB、IκB的表达均较模型组降低(P<0.05或P<0.01)；各治疗组大鼠子宫组织TNF-amRNA、ICAM-1mRNA表达水平均较模型组下降。其中盆腔炎中、大剂量组各指标水平除子宫组织ICAM-1mRNA均已接近正常对照组水平(P<0.05)。盆腔炎中剂量组NF-κB与TNF-amRNA表达、TNF-amRNA与TNF-a表达、NF-κB与ICAM-1mRNA表达、ICAM-1mRNA与ICAM-1表达之间均呈显著正相关性(P<0.01)。结论：活血补肾法通过对大鼠血清、子宫组织的炎症细胞因子和粘连相关指标表达的调节,恢复机体免疫平衡,有效抑制盆腔炎模型大鼠炎症,缓解盆腔粘连。活血补肾法调节机体免疫平衡的可能机制之一是通过抑制核因子的活化继而抑制其启动的多种细胞因子及免疫分子的转录及表达。更多还原

【Abstract】 Objective:To explore the immunoregulation mechanism of the treatment of promoting blood circulation and tonifying the kidney on the rats with chronic pelvic inflammatory disease(CPID) by observing the changes of inflammatory cytokines and adhesion related molecules and neuclear factor expression in serum or uterine tissue of CPID model rats.Method:Female Wistar rats were used and the CPID model was established with mixed bacteria injection combined with mechanic injury. Sixty-five rats were randomly divided into normal group, sham operation group, model group, Penqiangyan grannule high, medium and low dose groups and Penyanjing grannule group. The histomorphological changes of the rats’uteri were observed using light microscope. The enzyme-linked immunosorbent assay (ELISA) was applied to measure the concentration of pro-inflammatory cytokines (IL-1β, IL-8, MCP-1) and anti-inflammatory cytokines (IL-10) and transforming growth factor-β1(TGF-β1) in serum of rats in each group; the immunohistochemistry method was used to detect the expression of the following indices in rats’uterine tissue:tumor necrosis factor-α(TNF-α), intercellular adhesion molecule-1(ICAM-1), neuclear factor-kappaB(NF-κB) and inhibitor of nuclear factor-kappaB(IκB). The expression of TNF-αmRNA and ICAM-1mRNA were measured using RT-PCR.Results:The serous levels of pro-inflammatory cytokines (IL-1β, IL-8, MCP-1) and TGF-β1 level in model group were significantly higher than those in normal group (P<0.01); the serous levels of anti-inflammatory cytokines ((IL-4, IL-10) in model group were significantly lower than those in normal group. The expression of TNF-α, ICAM-1, NF-κB and IκB and the expression of TNF-αmRNA and ICAM-1mRNA were all significantly increased than those in normal group. After treatment, the pro-inflammatory cytokine levels were decreased while the anti-inflammatory cytokine levels were elevated. The expression of adhesion related immune molecules(TNF-α, ICAM-1) and NF-κB, IκB in the uterine tissue of model group was significantly decreased (P<0.01). Except for ICAM-1mRNA, all the other indices in Penqiangyan medium and large dose groups were similar to the levels of normal group (P>0.05).In Penqiangyan medium dose group, the expression of NF-κB, TNF-amRNA and TNF-a were correlated significantly, so were the expression of NF-κB, ICAM-1mRNA and ICAM-1.Conclusions:The method of promoting blood circulation and tonifying the kidney could adjust levels of the inflammatory cytokines and various adhesion-related molecules, thus restore the immune balance effectively so as to inhibit inflammation, prevent and alleviate the pelvic adhesions. The mechanism might be that the method could suppress the activation of NF-κB, therefore inhibit the transcription of certain inflammatory cytokines and immune-related adhesion molecule.更多还原