circZMYM2/miR-29a/PUMA轴对急性胰腺炎腺泡细胞凋亡的影响及作用机制  

Impact of the circZMYM2/miR-29a/PUMA axis on acinar cell apoptosis in acute pancreatitis and its action mechanism

作　　者：高明[1] 王琪 孙远松 李贺[1] GAO Ming;WANG Qi;SUN Yuansong;LI He(Department of Emergency Surgery,the Second Affiliated Hospital of Anhui Medical University,Hefei 230601,China)

机构地区：[1]安徽医科大学第二附属医院急诊外科,安徽合肥230601

出　　处：《中国普通外科杂志》2023年第9期1341-1348,共8页China Journal of General Surgery

基　　金：安徽医科大学校科学研究基金资助项目(2020xkj192)。

摘　　要：背景与目的:腺泡细胞凋亡在急性胰腺炎(AP)炎症反应和病情进展中发挥重要作用,进一步了解腺泡细胞的凋亡机制及相关通路有助于为AP特异性治疗提供新思路。本研究探讨锌指蛋白基因ZMYM2转录环状核糖核酸(circZMYM2)、微小RNA-29a(miR-29a)与p53上调凋亡调节因子(PUMA)在AP中的表达及其之间的潜在关系。方法:将大鼠胰腺腺泡细胞AR42J用雨蛙素诱导AP体外模型(AP组),或先通过pcDNA3.1-si-ZMYM2转染,敲低circZMYM2后再用雨蛙素诱导AP体外模型(si-circZMYM2组),以无处理的AR42J细胞作为对照组。3 h后分别用ELISA法检测细胞淀粉酶水平、CCK-8法检测细胞活力、流式细胞术与TUNEL法检测细胞凋亡情况、Western blot法检测细胞PUMA蛋白表达、qRT-PCR检测细胞circZMYM2与miR-29a表达。结果:与对照组比较,AP组与si-circZMYM2组淀粉酶水平均明显升高、细胞活力均明显降低、细胞凋亡率或凋亡细胞数均明显增加、PUMA蛋白表达水平均明显升高,但si-circZMYM2组以上指标的变化程度均明显低于AP组(均P<0.05)。与对照组比较,circZMYM2表达水平在AP组明显升高,在si-circZMYM2组明显降低,而miR-29a表达水平在AP组明显降低,在si-circZMYM2组明显升高(均P<0.05)。结论:circZMYM2在AP的腺泡细胞中表达升高,其可能通过与miR-29a内源性竞争作用,抑制miR-29a的表达,从而上调PUMA表达水平,促进腺泡细胞凋亡和AP进展。Background and Aims:Apoptosis of acinar cells plays a crucial role in the inflammatory response and progression of acute pancreatitis(AP).A better understanding of the apoptotic mechanisms and related pathways in acinar cells can provide new insights for AP-specific therapies.This study was conducted to investigate the expressions of the zinc finger protein gene ZMYM2 transcribed circular RNA(circZMYM2),microRNA-29a(miR-29a),and p53 upregulated modulator of apoptosis(PUMA)in AP and their potential relationships.Methods:Rat pancreatic acinar cells AR42J were induced to form an in vitro model of AP using cerulein(AP group)or were transfected with pcDNA3.1-si-ZMYM2 to knock down circZMYM2 expression before cerulein induction(si-circZMYM2 group),using untreated AR42J cells as a control group.After 3 h,the amylase level was determined by ELISA assay,the cell viability was assessed by CCK-8 assay,the apoptosis was measured by flow cytometry and TUNEL assay,and the PUMA protein expression was examined by Western blot analysis,and the circZMYM2 and miR-29a expression levels were detected by qRT-PCR method.Results:Compared to the control group,the AP group and si-circZMYM2 group showed significantly increased amylase levels,decreased cell viability,increased apoptosis rates or apoptotic cell numbers,and elevated PUMA protein expression(all P<0.05).However,the changes in the above indicators were significantly less pronounced in the si-circZMYM2 group than those in the AP group(all P<0.05).The expression level of circZMYM2 was significantly increased in the AP group and significantly decreased in the si-circZMYM2 group,while the expression level of miR-29a was significantly downregulated in the AP group and significantly upregulated in the si-circZMYM2 group compared to the control group(all P<0.05).Conclusion:The expression of circZMYM2 in acinar cells during AP is upregulated,and it may probably inhibit miR-29a expression through endogenous competition,thereby upregulate PUMA expression level and then promote acinar cell

关 键 词：胰腺炎 腺泡细胞 细胞凋亡 RNA 环状 微RNAS 

分 类 号：R657.5[医药卫生—外科学]