黄连调控miR-26a-3p对急性胰腺炎胰腺细胞增殖和凋亡的影响  

Effects of Rhizoma Coptis on the proliferation and apoptosis of pancreatic cells in acute pancreatitis by regulating miR-26a-3p in vitro

作　　者：曾小华 廖辉军[2] 刘柏京[3] 乐冬友[1] ZENG Xiaohua;LIAO Huijun;LIU Baijing(Emergency Department,Chenzhou First People’s Hospital,Hunan,Chenzhou 423000,China;不详)

机构地区：[1]湖南省郴州市第一人民医院急诊科,423000 [2]湖南省郴州市第一人民医院肝胆外科,423000 [3]湖南省郴州市第一人民医院疼痛科,423000

出　　处：《河北医药》2021年第10期1445-1450,共6页Hebei Medical Journal

基　　金：郴州市科技计划项目(编号:2019XJ88)。

摘　　要：目的探讨黄连提取物对急性胰腺炎(AP)胰腺细胞增殖和凋亡的影响及作用机制。方法利用雨蛙素作用AR42J细胞建立AP细胞模型。不同浓度的黄连提取物作用雨蛙素诱导的AR42J细胞,酶联免疫吸附法检测细胞培养上清中白细胞介素6(IL-6)和肿瘤坏死因子-α(TNF-α)水平,四甲基噻唑蓝染色法(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,蛋白印迹(Western Blot)检测P21、半胱天冬酶3(Caspases 3)蛋白表达水平,实时荧光定量PCR(RT-qPCR)检测miR-26a-3p表达水平。转染miR-26a-3p-mimics至AR42J细胞,相同方法观察过表达miR-26a-3p对雨蛙素诱导的AR42J细胞增殖、凋亡及炎性因子IL-6和TNF-α分泌的影响。双荧光素酶报告基因实验验证miR-26a-3p与髓系细胞触发受体1(TREM1)调控关系。结果黄连提取物或过表达miR-26a-3p均可降低雨蛙素诱导的AR42J细胞培养上清液中IL-6和TNF-α水平及细胞存活率(P<0.05),提高雨蛙素诱导的AR42J细胞凋亡率、P21和Caspase-3蛋白水平(P<0.05)。黄连提取物可促进雨蛙素诱导的AR42J细胞miR-26a-3p表达,抑制miR-26a-3p表达降低了黄连提取物对雨蛙素诱导的AR42J细胞存活率、凋亡率、P21和Caspase 3蛋白水平、IL-6和TNF-α水平的影响(P<0.05)。miR-26a-3p靶向负调控TREM1表达。TREM1过表达可降低黄连提取物对雨蛙素诱导的AR42J细胞存活率、凋亡率、P21和Caspase-3蛋白水平、IL-6和TNF-α水平的影响(P<0.05)。结论黄连提取物可降低雨蛙素诱导的AR42J细胞炎性反应及增殖,促进受损的细胞凋亡,其作用机制与调控miR-26a-3p/TREM1表达有关。Objective To investigate the effects and action mechanism of Rhizoma Coptidis extract on the proliferation and apoptosis of pancreatic cells in acute pancreatitis(AP).Methods AP cell model was established by using cerulein to treat AR42J cells,which were treated by different concentrations of Rhizoma Coptidis extracts.Then the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in cell culture supernatants were detected by enzyme linked immunosorbent assay(ELISA),the cell proliferation was detected by Tetramethylthiazole Blue staining(MTT),and cell apoptosis was detected by flow cytometry,the expression levels of P21 and caspase-3 were detected by Western Blot,and the miR-26a-3p expression level was detected by real time quantitative PCR(RT-qPCR).The miR-26a-3p mimics were transfected into AR42J cells.The same methods were used to observe the effects of overexpression of miR-26a-3p on the proliferation,apoptosis and the secretion of inflammatory factors IL-6 and TNF-α.Moreover dual luciferase reporter gene assay was used to identify the regulating relationship between miR-26a-3p and TREM1.Results The Coptidis Rhizome extract or overexpression of miR-26a-3p could significantly decrease the and cell viability and the levels of IL-6 and TNF-αin the culture supernatant of AR42J cells(P<0.05),significantly increase the apoptosis rate and expression levels of P21 and Caspase-3 protein(P<0.05).The Rhizoma Coptidis extract could promote the expression levels of miR-26a-3p in AR42J cells,and the inhibition of miR-26a-3p expression could decrease the effects of Coptidis Rhizome extract on the survival rate,apoptosis rate,the levels of P21,Caspase-3,IL-6 and TNF-αin AR42J cellscerulean(P<0.05).MiR-26a-3p targetedly negatively regulated the TREM1 expression.Moreover the overexpression of TREM1 could decrease the effects of Coptidis Rhizome extract on the survival rate,apoptosis rate,and the levels of P21,Caspase-3,IL-6 and TNF-αin AR42J cells(P<0.05).Conclusion Coptidis Rhizoma extract can reduce the inflamm

关 键 词：黄连提取物 急性胰腺炎 miR-26a-3p 髓系细胞触发受体1 细胞增殖 凋亡 

分 类 号：R657.51[医药卫生—外科学]