宁泌泰胶囊通过调节Nrf2和NF-κB信号通路减轻慢性前列腺炎/慢性盆腔疼痛综合征大鼠的症状

Ningmitai Capsules alleviate CP/CPPS symptoms by regulating Nrf2 and NF-κB signaling pathways

目的:评价宁泌泰胶囊治疗大鼠慢性前列腺炎/慢性盆腔疼痛综合征(CP/CPPS)的疗效及机制。方法:6~8周龄雄性Wistar大鼠15只,将大鼠随机分为假手术组、模型组对照组和宁泌泰治疗组,每组5只。假手术组大鼠前列腺腹叶处注射无菌PBS,术后第2天起施用纯净水灌胃(3 ml/d);模型对照组大鼠前列腺腹叶处注射50μl完全弗氏佐剂(CFA),术后第2天起施用纯净水灌胃(3 ml/d);宁泌泰治疗组大鼠前列腺腹叶处注射50μl CFA,术后第2天起施用3 ml宁泌泰[400 mg/(kg·d)]混悬液灌胃。4周后处死大鼠获取血清和前列腺组织样本,通过HE染色采用慢性前列腺炎的组织病理学分级系统评估前列腺炎的严重程度,通过实时定量PCR检测组织炎症因子的mRNA表达水平,通过试剂盒检测相关抗氧化酶活力,通过Western印迹实验检测信号通路相关靶点的表达水平。结果:组织形态学分析发现模型对照组间质有不同程度的炎性细胞浸润、炎性空泡、不规则状腺泡及水肿表现,与假手术组相比炎症评分显著增加(P<0.05),宁泌泰治疗组中浸润淋巴细胞和炎性空泡减少,与模型对照组相比炎症评分显著降低(P<0.05);与假手术组相比,模型对照组中炎症因子相关基因IL-1β、IL-6、IL-10、IL-17A、TNFα、IFNγ mRNA表达水平显著上调(P<0.05),超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)含量显著降低(P<0.05),丙二醛(MDA)表达水平显著升高(P<0.05);经过宁泌泰治疗后与模型对照组相比,宁泌泰治疗组中炎症因子相关基因IL-1β、IL-6、IL-10、IL-17A、TNFα、IFNγ mRNA表达水平显著降低(P<0.05),SOD、CAT、GSH-Px含量显著升高(P<0.05),MDA表达水平显著降低(P<0.05),核因子红系2相关因子2 (n-Nrf2)、血红素氧合酶1 (HO-1)的表达显著上调(P<0.05),NF-κB p-p65的表达显著下调(P<0.05)。结论:宁泌泰能够在大鼠CP/CPPS的发病机制中发挥抗炎和抗氧化应激作用,其机制可能通过激活Nrf2和抑制NF-κB信号通路实现。

Objective:To investigate the therapeutic effect of Ningmitai Capsules(NMT) on chronic prostatitis/chronic pelvic pain syndrome(CP/CPPS) and its action mechanism.Methods:We equally randomized fifteen 6-8 weeks old male Wistar rats into a sham operation,a model control and an NMT group,and established a CP/CPPS model in the rats by intra-prostatic injection of complete Freund adjuvant(CFA).We treated the rats in the sham operation and model control groups by injection of aseptic phosphate buffer saline(PBS) and 50 μl complete Freund adjuvant(CFA),respectively,into the ventral lobe of the prostate followed by gavage with purified water from the second day after surgery,and those in the NMT group with 50 μl CFA injected into the ventral lobe of the prostate followed by gavage with 3 ml NMT suspension at 400 mg/kg/d from the second day after surgery.After 4 weeks of treatment,we killed the animals and collected the serum and prostatic tissue samples for evaluation of the severity of prostatitis by histopathological grading of chronic prostatitis through HE staining,determination of the mRNA expressions of inflammatory factors by real-time quantitative PCR,measurement of the activities of related antioxidant enzymes with testing kits,and detection of the expressions of the related targets in the signaling pathways by Western blot.Results:Histopathological examination revealed different degrees of inflammatory cell infiltration in the mesenchyme,inflammatory vacuoles,irregularly shaped acini in the model control group,with a significantly higher inflammation score than in the sham operation group(P<0.05),and reduced infiltrating lymphocytes,inflammatory vacuoles and inflammation score in the NMT group compared with those in the model controls(P<0.05).Compared with the rats in the sham operation group,the model controls showed remarkably up-regulated expressions of IL-1β,IL-6,IL-10,IL-17A,TNF-α and IFN-γ(P<0.05),decreased contents of superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSH-Px)(P<0.05),and increased content of malondialdehyde(MDA)(P<0.05).Compared with the model controls,the rats treated with NMT exhibited markedly reduced expressions of IL-1β,IL-6,IL-10,IL-17A,TNF-α and IFN-γ(P<0.05),increased contents of SOD,CAT and GSH-Px(P<0.05),decreased content of MDA(P<0.05),up-regulated expressions of nuclear factor red 2-related factor 2(n-Nrf2) and heme oxygenase-1(HO-1)(P<0.05),and down-regulated expression of NF-κB P-P65(P<0.05).Conclusion:NMT plays anti-oxidation and anti-inflammation roles in the pathogenesis of CP/CPPS,possibly by activation of Nrf2 and suppression of NF-κB signaling pathways.