摘要
目的基于NF-κB/IL-6信号通路研究雷公藤甲素对雨蛙素诱导的慢性胰腺炎(CP)小鼠模型胰腺纤维化的影响。方法随机将15只C57BL/6雄性小鼠分为对照组(生理盐水处理,n=5)、雨蛙素组(雨蛙素诱导CP模型,n=5)和雷公藤甲素组(雨蛙素诱导+雷公藤甲素治疗,n=5)。6周后采集样本进行检测。对小鼠胰腺称重;苏木素-伊红染色、天狼猩红及马松染色观察胰腺组织形态及胶原沉积;ELISA检测血清中IL-6的表达;免疫组化检测α平滑肌肌动蛋白(α-SMA)、NF-κB/p65的表达情况;蛋白质免疫印迹检测NF-κB/p65、IL-6及α-SMA的表达。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果3组间胰腺组织重量,IL-6水平,病理评分,天狼猩红染色、马松染色、α-SMA阳性信号、NF-κB/p65阳性信号的平均光密度值比较差异均有统计学意义(F值分别为64.87、15.85、145.33、141.80、121.77、250.22、69.22,P值均<0.001)。与雨蛙素组相比,雷公藤甲素组胰腺重量明显增加,IL-6的表达明显降低,胰腺组织纤维化及胶原沉积明显减弱,α-SMA与NF-κB/p65表达量降低(P值均<0.05)。蛋白质免疫印迹检测结果显示,3组间NF-κB/p65、IL-6、α-SMA表达水平比较差异均有统计学意义(F值分别为8.86、6.74、16.23,P值均<0.05),雷公藤甲素组胰腺组织NF-κB/p65、IL-6及α-SMA表达量明显低于雨蛙素组(P值均<0.05)。结论雷公藤甲素可抑制NF-κB/p65及IL-6蛋白的表达,缓解雨蛙素诱导的CP小鼠模型的胰腺纤维化。
Objective To investigate the effect of triptolide on fibrosis in mice with cerulein-induced chronic pancreatitis(CP)based on the nuclear factor-kappa B(NF-κB)/interleukin-6(IL-6)signaling pathway.Methods A total of 15 male C57BL/6 mice were randomly divided into control group(treated with normal saline),cerulein group(model mice with cerulein-induced CP),and triptolide group(induced by cerulean and treated with triptolide),with 5 mice in each group.Samples were collected for detection after 6 weeks.The weight of the pancreas was measured;HE staining,picrosirius red staining,and Masson staining were used to observe pancreatic histomorphology and collagen deposition;ELISA was used to measure the expression of IL-6 in serum;immunohistochemistry was used to measure the expression of alpha-smooth muscle actin(α-SMA)and NF-κB/p65;Western blot was used to measure the expression of NF-κB/p65,IL-6,andα-SMA.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results There were significant differences between the three groups in the weight of the pancreas,IL-6 concentration,pathological score,picrosirius red staining results,Masson staining results,and mean optical density of positiveα-SMA signal and positive NF-κB/p65 signal(F=64.87,15.85,145.33,141.80,121.77,250.22,and 69.22,all P<0.001).Compared with the cerulein group,the triptolide group had a significant increase in the weight of the pancreas and significant reductions in the expression of IL-6,pancreatic fibrosis,collagen deposition,and the expression ofα-SMA and NF-κB/p65(all P<0.05).Western blotting showed that there were significant differences in the expression of NF-κB/p65,IL-6,andα-SMA between the three groups(F=8.86,6.74,and 16.23,all P<0.05),and compared with the cerulein group,the triptolide group had significantly lower expression of NF-κB/p65,IL-6,andα-SMA in the pancreatic tissue(all P<0.05).Conclusion Triptolide alleviates fibrosis in mice with cerulein-induced CP and inhibit the protein expression of NF-κB/p65 and IL-6.
作者
谭鹏
陈浩
王安康
钱保林
黄治伟
石昊
高琳
付文广
TAN Peng;CHEN Hao;WANG Ankang(Academician Expert Workstation of Sichuan Province,The Affiliated Hospital of Southwest Medical University,Luzhou,Sichuan 646000,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2020年第3期641-645,共5页
Journal of Clinical Hepatology
基金
四川省应用基础研究项目(2018JY0283)
四川省卫生健康委员会科研课题(19PJ292)。