连翘对重症急性胰腺炎中炎症因子及细胞自噬的影响

Effects of forsythia forsythia on inflammatory factors and autophagy in severe acute pancreatitis

目的:探讨重症急性胰腺炎(SAP)细胞自噬的变化以及连翘干预后对细胞自噬及炎症因子的影响。方法:SD雄性大鼠36只,随机分成对照组、SAP组及连翘干预组。其中连翘干预组又分为连翘高浓度(AFS1)、连翘中浓度(AFS2)、连翘低浓度(AFS3)组(分别对应5 g/kg、2.5 g/kg、1.25 g/kg)及阳性对照组。经胰胆管逆行注射5%牛黄胆酸钠溶液造模,造模后12 h处死大鼠,留取标本,测量并比较各组肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)、血淀粉酶(AMY),免疫组化观察蛋白轻链3(LC3II)、自噬相关蛋白p62、溶酶体相关膜蛋白2(LAMP2)的表达水平,胰腺标本进行病理观察及评分。结果:与对照组相比,SAP组AMY、IL-1β、TNF-α水平均升高(P<0.05),免疫组化观察LC3II、P62表达增加(P<0.05),LAMP2的表达降低(P<0.05),与SAP组相比,连翘干预组IL-1β、TNF-α、AMY水平均降低(P<0.05),且存在明显的量效关系;胰腺组织炎症也明显减轻;而且LC3II、LAMP2表达增加(P<0.05),P62的表达降低(P<0.05)。结论:连翘能显著降低重症急性胰腺炎时TNF-α、IL-1β等炎症因子的释放,修复受损自噬,从而减轻重症急性胰腺炎,其可能通过核因子-кB信号通路发挥作用。

Objective To investigate the pancreophagy changes in patients with severe acute pancreatitis(SAP)and the effects of forsythiae intervention on autophagy and inflammatory factors.Methods 36 male SD rats were randomly divided into control group,SAP group and forsythia intervention group.Forsythia intervention group was further divided into AFS1,AFS2,AFS3 groups(corresponding to 5 g/kg,2.5 g/kg and 1.25 g/kg respectively)and positive control group.The rats were sacrificed 12 hours after modeling by retrograde injection of 5%sodium taurine cholic solution.Samples were collected and the expressions of TNF-α,IL-1βand blood amylase(AMY)in each group were measured.The expressions of LC3II,P62 and LAMP2 were observed by immunohistochemistry.Results Compared with the control group,the levels of AMY,IL-1βand TNF-αin SAP group were increased(P<0.05),the expressions of LC3II and P62 were increased(P<0.05),and the expression of LAMP2 was decreased(P<0.05).Compared with SAP group,the levels of IL-1β,TNF-αand AMY in the forsythia intervention group were decreased(P<0.05),and there was a significant dose-effect relationship.The inflammation of pancreatic tissue was also significantly reduced.The expressions of LC3II and LAMP2 were increased(P<0.05),while the expression of P62 was decreased(P<0.05).Conclusion Forsythia can significantly reduce the release of TNF-α,IL-1βand other inflammatory cytokines in severe acute pancreatitis,repair damaged autophagy and alleviate severe acute pancreatitis,which may play a role through the nuclear factor-кB signaling pathway.