柴黄清胰活血颗粒对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的保护作用及机制  

Investigate the protective effect and mechanism of Chaihuang Qingyi Huoxue Granule on pancreatic acinous cell injury induced by caerulin

作　　者：付加伟 唐祖鑫 陆俊 邹鑫 李丽 赵龙 李志 FU Jiawei;TANG Zuxin;LU Jun;ZOU Xin;LI Li;ZHAO Long;LI Zhi(College of Integrated Traditional Chinese and Western Medicine,Southwest Medical University,Luzhou 646000,China;Department of Spleen and Stomach Diseases,The Affiliared Traditional Chinese Medicine Hospital,Southwest Medical University,Luzhou 646000,China;The Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Digestive System Diseases of Luzhou city,Luzhou 646000,China)

机构地区：[1]西南医科大学中西医结合学院,泸州646000 [2]西南医科大学附属中医医院脾胃病科,泸州646000 [3]泸州市中西医结合防治消化系统疾病重点实验室,泸州646000

出　　处：《西南医科大学学报》2024年第3期251-255,共5页Journal of Southwest Medical University

基　　金：四川省科技厅项目(2019YFS0163);四川省中管局项目(2023MS416);川大-泸州市政府战略合作项目(2020CDLZ-18);西南医科大学校级课题(2022QN095);西南医科大学附属中医医院科研团队创新培育项目(2022-CXTD-01)。

摘　　要：目的观察柴黄清胰活血颗粒对雨蛙素(caerulein)诱导的大鼠胰腺腺泡细胞损伤的保护作用,并探讨柴黄清胰活血颗粒对JAK2/STAT3信号通路的调控机制。方法取对数生长期的AR42J细胞,通过CCK-8法检测不同浓度柴黄清胰活血颗粒对AR42J细胞活性的影响并筛选最佳药物干预浓度;用雨蛙素刺激细胞6 h构建胰腺炎体外模型;随机分为空白对照组、模型组、柴黄清胰活血颗粒低、中、高剂量组(0.5、1、2 mg/mL);ELISA法测定细胞上清液中TNF-α、IL-6、IL-1β的表达;实时荧光定量PCR测定各组细胞JAK2、STAT3基因表达水平;Western blot法检测各组细胞JAK2、STAT3、p-JAK2、p-STAT3蛋白表达水平。结果CCK-8法检测发现,当药物浓度为0.5、1及2 mg/mL时,平均细胞存活率均大于100%;ELISA检测发现,与空白组相比,模型组炎性细胞因子IL-6、IL-8、TNF-α升高,提示AR42J细胞急性胰腺炎模型的成功建立;与模型组相比,柴黄清胰活血颗粒组(0.5、1、2 mg/mL)IL-6、IL-1β、TNF-α明显降低,差异具有统计学意义(P<0.05);实时荧光定量PCR检测发现,与模型组相比,柴黄清胰活血颗粒组(0.5、1、2 mg/mL)STAT3 mRNA、柴黄清胰活血颗粒组(2 mg/mL)JAK2 mRNA表达水平显著下降(P<0.05);westen blot测定发现,与模型组相比,柴黄清胰活血颗粒组(0.5、1、2 mg/mL)p-JAK2/JAK2、p-STAT3/STAT3蛋白相对表达量显著下降(P<0.05)。结论柴黄清胰活血颗粒可能通过调控JAK2/STAT3信号通路中关键分子表达,从而减轻AR42J细胞炎症反应。Objective To investigate the protective effect and underlying mechanism of Chaihuang Qingyi Huoxue Granule on the JAK2/STAT3 signal pathway in rat pancreatic acinar cells induced by caerulin.Methods The pancreatic acinar cells AR42J were cul⁃tured and treated with different concentrations of Chaihuang Qingyi Huoxue granules.The effects of different concentrations of Chai⁃huang Qingyi Huoxue granules on the viability of AR42J cells were detected by CCK-8 assay to determine the best drug concentration.AR42J cells were stimulated with caerulin for 6 hours to mimic acute pancreatitis and randomly divided into the control group,the model group,the low,middle,and high dose groups of Chaihuang Qingyi Huoxue Granule.The contents of TNF-α,IL-6,and IL-1βin culture supernatants were determined by ELISA;The expression of JAK2 and STAT3 genes were detected by real-time quantitative PCR(q-RTPCR);The protein expression levels of JAK2,STAT3,p-JAK2,p-STAT3 were detected by Western blot.All the above da⁃tas were analyzed using GraphPad Prism 9.0 software and one-way analysis of variance.Results CCK-8 assay showed that the average cell survival rate was more than 100%when the drug concentration were 0.5,1,and 2 mg/L.ELISA showed that compared with the con⁃trol group,the inflammatory cytokines IL-6,IL-8,and TNF-αwere increased in the model group,suggesting the successful establish⁃ment of acute pancreatitis.The contents of TNF-α,IL-6,and IL-1βin cell supernatants of AR42J cells treated with Chaihuang Qingyi Huoxue Granule group(0.5,1,2 mg/mL)were significantly lower than those in the model group after treatment(P<0.05).The con⁃tents of STAT3 mRNA in AR42J cell in the Chaihuang Qingyi Huoxue Granule group(0.5,1,2 mg/mL),JAK2 mRNA in AR42J cell in the Chaihuang Qingyi Huoxue Granule group(2 mg/mL)were significantly lower than those in the model group after treatment(P<0.05).The ratio of p-JAK2/JAK2,p-STAT3/STAT3 in AR42J cell in the Chaihuang Qingyi Huoxue Granule group(0.5,1,2 mg/mL)were significantly lower than thos

关 键 词：柴黄清胰活血颗粒 JAK2/STAT3信号通路 AR42J细胞 急性胰腺炎 

分 类 号：R285.5[医药卫生—中药学]