紫草素对大鼠重症急性胰腺炎的保护作用及其对TLR4/NF-κB信号通路的影响

Protective effect of shikonin on severe acute pancreatitis in rats and its effect on TLR4/NF-κB signaling pathway

目的:研究紫草素对大鼠重症急性胰腺炎的保护作用及其对Toll样受体4/核因子-κB(TLR4/NF-κB)信号通路的影响,探讨紫草素保护大鼠重症急性胰腺炎的可能机制。方法:将100只大鼠根据随机数字法分为对照组、模型组、紫草素1组和紫草素2组,每组25只。牛磺脱氧胆酸钠建立重症急性胰腺炎大鼠模型,紫草素1组和紫草素2组大鼠分别于术前1 d分别给予紫草素1 mg/kg和2 mg/kg腹腔注射。计算胰腺重量指数。酶联免疫吸附试验(ELISA)测定血清脂肪酶、淀粉酶水平。伊红-苏木素(HE)染色测定胰腺组织病理形态学变化。Western blot测定胰腺组织中IL-6、TNF-α、NF-κBp65和核转录因子κB抑制蛋白-α(IκB-α)蛋白水平。免疫组化法测定胰腺组织中NF-κBp65蛋白表达。结果:与对照组比较,模型组、紫草素1组、紫草素2组大鼠胰腺重量指数、血脂肪酶和淀粉酶水平、胰腺病理损伤指数、胰腺组织中IL-6、TNF-α、NF-κBp65蛋白水平、NF-κBp65阳性细胞率升高(P<0.05),胰腺组织中IκB-α蛋白水平降低(P<0.05);与模型组比较,紫草素1组和紫草素2组大鼠胰腺重量指数、血脂肪酶和淀粉酶水平、胰腺病理损伤指数、胰腺组织中IL-6、TNF-α、NF-κBp65蛋白水平、NF-κBp65阳性细胞率降低(P<0.05),胰腺组织中IκB-α蛋白水平升高(P<0.05);与紫草素1组比较,紫草素2组大鼠胰腺重量指数、血脂肪酶和淀粉酶水平、胰腺病理损伤指数、胰腺组织中IL-6、TNF-α、NF-κBp65蛋白水平、NF-κBp65阳性细胞率降低(P<0.05),胰腺组织中IκB-α蛋白水平升高(P<0.05)。结论:紫草素可能通过TLR4/NF-κB信号通路发挥大鼠重症急性胰腺炎的保护作用。

Objective:To study the protective effect of shikonin on severe acute pancreatitis in rats and its effect on Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway,and to explore the possible mechanism of shikonin in protecting severe acute pancreatitis in rats.Methods:100 rats were divided into control group,model group,shikonin group 1 and shikonin group 2 accorded to random number method,25 rats in each group.Snail sodium taurodeoxycholate was used to establishes the rat model of severe acute pancreatitis.Rats in shikonin group 1 and shikonin group 2 were given intraperitoneal injection of shikonin 1 mg/kg and 2 mg/kg,respectively at 1 day before surgery.The pancreas weight indexs were calculated.The serum lipase and amylase levels were determined by enzyme-linked immunosorbent assay(ELISA).The pathological changes of pancreatic tissue were determined by hematoxylin-eosin(HE)staining.The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),NF-κBp65 and nuclear factor kappa B inhibitory protein-α(IκB-α)protein in pancreatic tissue were determined by Western blot.The expression of NF-κBp65 protein in pancreatic tissue was determined by immunohistochemistry.Results:Compared with the control group,the pancreatic weight index,blood lipase and amylase levels,pancreatic pathological injury index,the IL-6,TNF-αand NF-κBp65 protein levels in pancreatic tissue,and the rate of NF-κBp65 positive cells were increased(P<0.05),and the level of IκB-αprotein in pancreatic tissues were decreased(P<0.05)in model group,shikonin group 1 and shikonin group 2.Compared with the model group,the pancreatic weight index,blood lipase and amylase levels,pancreatic pathological injury index,the IL-6,TNF-αand NF-κBp65 protein levels in pancreatic tissue,and the rate of NF-κBp65 positive cells were decreased(P<0.05),and the level of IκB-αprotein in pancreatic tissues were increased(P<0.05)in shikonin group 1 and shikonin group 2.Compared with shikonin group 1,the pancreatic weight index,blood lipase and amylase levels,pancreatic pathological injury index,the IL-6,TNF-αand NF-κBp65 protein levels in pancreatic tissue,and the rate of NF-κBp65 positive cells were decreased(P<0.05),and the level of IκB-αprotein in pancreatic tissues were increased(P<0.05)in shikonin group 2.Conclusion:Shikonin may play a protective role in severe acute pancreatitis in rats through the TLR4/NF-κB signaling pathway.