黄连素对重症急性胰腺炎大鼠炎症反应的影响

Effect of berberine on the inflammatory response in rats with severe acute pancreatitis

目的探讨黄连素对重症急性胰腺炎大鼠的保护作用及机制。方法 30只雄性SD大鼠按双盲法随机均分为对照组(Sham组)、重症急性胰腺炎模型组(AP组)和黄连素组(BBR组),每组10只。将牛磺胆酸钠注射入大鼠胰胆管的方法构建重症急性胰腺炎大鼠模型。BBR组在建模前5 d每天给予黄连素灌胃1次,Sham组和AP组术前5 d每天给予等量生理盐水灌胃。建模后24 h,处死大鼠收集胰腺和血清。应用酶联免疫吸附测定检测血清中淀粉酶、脂肪酶、白介素-6(IL-6)、白介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)表达水平;应用反转录PCR检测胰腺组织中IL-6、IL-1β和TNF-α的信使RNA(m RNA)水平;应用蛋白质印迹法检测胰腺跨膜蛋白4(TLR4)、髓样分化因子88(MyD88)、磷酸化p65(p-p65)和p65表达水平。结果 AP组大鼠的淀粉酶、脂肪酶、IL-6、IL-1β、TNF-α、TLR4、MyD88和p-p65表达水平均较Sham组明显增高,差异均有统计学意义(P<0.05),但p65表达水平两组比较差异无统计学意义(P>0.05);与模型AP组相比,BBR组大鼠的淀粉酶、脂肪酶、IL-6、IL-1β、TNF-α、TLR4、MyD88和p-p65表达水平均明显降低,差异均有统计学意义(P<0.05),但p65表达水平两组比较差异无统计学意义(P>0.05);与Sham组相比,BBR组大鼠的淀粉酶、脂肪酶、IL-6、IL-1β、TNF-α、TLR4、MyD88、p-p65表达差异均无统计学意义(P>0.05)。结论黄连素可通过抑制炎症反应而减轻大鼠重症急性胰腺炎的发生,其作用机制与抑制TLR-4/NF-κB信号通路激活相关。

Objective To investigate the protective effect and mechanism of berberine on rats with severe acute pancreatitis. Methods Thirty male SD rats were randomly divided into three groups： the control group（Sham group）, the severe acute model group（AP group） and the berberine group（BBR group） by double blind method, with 10 rats in each group. The severe acute pancreatitis model was constructed by injecting sodium taurocholate into the pancreatic duct in rats. Rats in BBR group were lavaged with berberine（1 time/d） every day for 5 days before modeling, and the those in Sham group and AP group were lavaged with the same amount of normal saline every day for 5 days before operation. The rats were sacrificed to collect pancreas and serum 24 h after model establishment. Enzyme-linked immunosorbent assay（ELISA） was used to detect the expression of amylase, lipase, interleukin-6（IL-6）, interleukin 1 beta（IL-1 β）, tumor necrosis factor α（TNF-α）. Reverse transcription PCR（RT-PCR） was used to detect the messenger m RNA level of IL-6, IL-1β and TNF-α in pancreatic tissues. Western blotting was used to examine the expression level of To-lllike receptor 4（TLR4）, myeloiddifferentiationfactor88（My D88）, phosphorylated p65（p-p65） and p65 in pancreatic tissues. Results The expression of amylase, lipase, IL-6, IL-1β, TNF-α, TLR4, My D88 and p-p65 in AP group were significantly higher than those in Sham group（P0.05）. However, there were no significant differences in p65 expression between the two groups（P0.05）. The expression of amylase, lipase, IL-6, IL-1β, TNF-α, TLR4, My D88 and p-p65 in BBR group were significantly lower than those in AP group（P0.05）. However, there were no significant differences in p65 expression between the two groups（P0.05）. There were no significant differences in the expression of amylase, lipase, IL-6, IL-1β, TNF-α, TLR4, My D88, and p-p65 between BBR group and Sham group（P0.05）.Conclusion Berberine could alleviate severe acute pancreatitis in rats by inhibiting the inflammatory reaction, and its mechanism is associated with suppressing the activation of TLR-4/NF-κB signal pathway.