基于RIPK1/RIPK3/MLKL信号通路探讨清解化攻方对重症急性胰腺炎大鼠胰腺坏死性凋亡的影响  

Effect of Qingjie Huagong decoction on pancreatic necrotic apoptosis in rats with severe acute pancreatitis based on RIPK1/RIPK3/MLKL signaling pathway

作　　者：朱晓东 冯敏超 刘锟荣 班莹 唐曦平 陈国忠[2,4] ZHU Xiao-dong;FENG Min-chao;LIU Kun-rong;BAN Ying;TANG Xi-ping;CHEN Guo-zhong(Guangxi University of Chinese Medicine,Nanning 530200,China;the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530023,China;Guangxi Medical University Affiliated Cancer Hospital,Nanning 530021,China;Guangxi Key Laboratory of Molecular Biology for the Prevention and Treatment of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广西中医药大学第一附属医院,广西南宁530023 [3]广西医科大学附属肿瘤医院,广西南宁530021 [4]广西中医药防治医学分子生物重点实验室,广西南宁530023

出　　处：《中国药理学通报》2024年第8期1489-1494,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.82160890);广西自然科学基金面上项目(No.2024GXNSFAA010246);国家中医优势专科建设项目(No.2024010);广西中医药大学引进博士科研启动基金项目(No.2022BS029);广西自然科学基金面上项目(No.2020GXNSFAA297062)。

摘　　要：目的探讨清解化攻方调控RIPK1/RIPK3/MLKL信号通路对重症急性胰腺炎(severe acute pancreatitis,SAP)大鼠胰腺坏死性凋亡的治疗作用及机制。方法采用逆行胰胆管注射牛磺胆酸钠建立SAP大鼠模型,分别设置假手术组、模型组、不同剂量清解化攻方给药组和阳性对照组,不同剂量清解化攻方组予低中高剂量中药灌胃、阳性对照组予乌司他丁药物干预、假手术组和模型组予生理盐水灌胃;HE染色观察胰腺病理;ELISA检测大鼠血清α-淀粉酶、IL-1β、IL-6和TNF-α水平;免疫组化和Western blot检测大鼠胰腺组织RIPK1、RIPK3、MLKL蛋白表达;qRT-PCR检测大鼠胰腺组织RIPK1、RIPK3、MLKL mRNA转录水平。结果与假手术组比,模型组大鼠胰腺针叶细胞弥漫性坏死、小叶间隔水肿明显、炎性细胞浸润,α-淀粉酶、IL-1β、IL-6和TNF-α水平明显升高(P<0.01),RIPK1、RIPK3、MLKL蛋白及mRNA表达水平显著升高(P<0.01);与模型组比,清解化攻方各剂量组和阳性对照组明显改善胰腺组织病理学,减少胰腺组织坏死凋亡,降低α-淀粉酶、IL-1β、IL-6和TNF-α表达水平(P<0.01),降低RIPK1、RIPK3、MLKL蛋白及mRNA表达水平(P<0.01)。结论清解化攻方可能通过调控RIPK1/RIPK3/MLKL信号通路,改善胰腺组织坏死性凋亡,从而起到保护胰腺组织的作用,减缓疾病的进展。Aim To explore the therapeutic effect of Qingjie Huagong decoction in regulating RIPK1/RIPK3/MLKL signaling pathway on pancreatic necrotic apoptosis in rats with severe acute pancreatitis(SAP)and the underlying mechanism.Methods The SAP rat model was established by retrograde pancreaticobiliary injection of sodium taurocholate,and the sham-operation group,the model group,the group with different dosages of Qingjie Huagong decoction and the positive control group were set up respectively.The group with different dosages of Qingjie Huagong decoction was given low,medium and high dosages of traditional Chinese medicine in the gastric gavage,the positive control group was given ulinastatin drug intervention,and the sham-operation and the model group were given physiological saline in the gastric gavage;HE staining was applied to observe pancreatic pathology;ELISA was used to measure the serum levels ofα-amylase,IL-1β,IL-6,and TNF-α;immunohistochemistry and Western blot were employed to determine the RIPK1,RIPK3,MLKL protein expression in rat pancreatic tissue;and qRT-PCR was utilized to detect the transcription levels of RIPK1,RIPK3 and MLKL mRNA in rat pancreatic tissue.Results Compared with the sham-operated group,the model group showed diffuse necrosis of pancreatic acinar cells,obvious interlobular septal edema,inflammatory cell infiltration,significantly higher levels ofα-amylase,IL-1β,IL-6,and TNF-α(P<0.01),and significantly higher expression levels of RIPK1,RIPK3,and MLKL proteins and mRNAs(P<0.01)in the model group;compared with the model group,the Qingjie Huagong decoction dose groups and positive control group significantly improved pancreatic histopathology,reduced pancreatic tissue necrosis and apoptosis,lowered the expression levels ofα-amylase,IL-1β,IL-6 and TNF-α(P<0.01),and reduced the expression levels of RIPK1,RIPK3,and MLKL proteins and mRNAs(P<0.01).Conclusions Qingjie Huagong decoction may improve the necrotic apoptosis of pancreatic tissue by regulating the RIPK1/RIPK3/MLKL signaling pa

关 键 词：重症急性胰腺炎 清解化攻方 RIPK1/RIPK3/MLKL信号通路 坏死性凋亡 胰腺 作用机制 

分 类 号：R-332[医药卫生] R285.5R322.491R329.25R576.1