微小RNA-92b在大鼠急性胰腺炎中对腺泡细胞凋亡的影响

The influence of microRNA-92b on apoptosis of pancreatitic acinus cells in rats with acute pancrea- tiffs

目的观察微小RNA-92b（miRNA-92b）在大鼠急性胰腺炎中的表达及其对急性胰腺炎腺泡细胞凋亡的影响。方法以50μg/L肿瘤坏死因子（TNF）-α诱导大鼠胰腺腺泡细胞（AR42J）建立体外急性水肿型胰腺炎模型，未处理的AR42J作为对照，检测干预12h后培养基中淀粉酶含量，实时荧光定量聚合酶链反应（FQ—PCR）检测miRNA-92b在1、3、6h时表达量变化。慢病毒携带miRNA-92b的模拟物（miRNAmimic）及反义寡核苷酸链（miRNAAMO）转染AR42J，转染空病毒组作为对照。建立体外急性胰腺炎模型后，使用FQ-PCR检测miRNA-92b表达量；流式细胞术检测AR42J细胞凋亡率，Westernblot法检测活性半胱氨酰天冬氨酸特异性蛋白酶3（Caspase-3）表达量变化。结果TNF—α.诱导AR42J建立的体外急性胰腺炎模型，miRNA-92b3h表达水平较对照组升高（1．81±0．09）倍（P〈0．05）；转染miRNA-92bmimic组miRNA-92b表达量较空病毒组升高（3．71±0．33）倍（P〈0．05），AR42J凋亡率[（41．20±2．42）％]较空病毒组[（23．50±1．85）％]明显升高（P〈0．05），活性Caspase-3表达量明显升高；转染miRNA-92bAMO组较空病毒组miRNA-92b表达量降低（O．58±0．15）倍（P〈0．05），AR42J凋亡率[（13．80±0．40）％]较空病毒组[（23．50±1．85）％]明显降低（P〈0．05），活性Caspase-3表达量明显降低。转染空病毒组较未转染组miRNA-92b表达量、细胞凋亡率、活性Caspase-3表达量差异均无统计学意义（P〉0．05）。结论急性水肿性胰腺炎发生时miRNA-92b表达量明显升高；上调miRNA-92b的表达，胰腺腺泡细胞的凋亡率增高，下调其表达胰腺腺泡细胞的凋亡率降低。

Objective To analyze the expression of microRNA-92b （miRNA-92b） in rats with a- cute pancreatitis and the influence of its expression change on apoptosis of pancreatic acinus cells in rats with acute pancreatitis. Methods An acute edema pancreatitis （AEP） model was established by inducing the rat pancreatic acinus cell line （AR42J） with 50 μg/L recombinant rat tumor necrosis factor （TNF）-α. Amylase was determined. The expression of miRNA-92b was detected by using real-time fluorescent quanti- tative polymerase chain reaction （FQ-PCR）. Lentivirus, which carrying the miRNA mimic and anti-miRNA oligonucleotide of the miRNA-92b, was used to transfect the AR42J cells. The AR42J cells, which were transfected with vehicle control, served as the control group. After transfection FQ-PCR was performed to detect the miRNA-92b expression. The flow cytometry was used to detect the apoptosis of AR42J cells after establishing the AEP model. The expression of activated Caspase-3 was detected by using Western blotting. Results The expression of miRNA 92b in the AR42J cells was increased by （ 1.81± 0. 09 ） times in the experimental group as compared with the control groups （ P 〈 0. 05 ） after establishing the AEP model. The miRNA-92b level was elevated by （3.71 ±0. 33 ） times after AR42J cells were transfected with miRNA-92b mimic as compared with the vehicle groups （ P 〈 0.05 ）. The AR42J cells showed more tendency to apopto- sis [ （41.20± 2.42 ） % ] than the vehicle groups [ （ 23.50 ± 1.85 ） %, P 〈 0. 05 ] and the level of Caspase-3 was increased obviously. The AR42J cells that were transfected with miRNA-92b AMO expressed the lower level of miRNA-92b about （ 0. 58 ± 0. 15 ） times and less apoptosis [ （ 13.80 ± 0.40） % ] than the vehicle groups （ P 〈 0. 05 ）. Conclusion The level of miRNA-92b was increased in AEP. The up-regulated expression of miRNA-92b may increase the apoptosis level of AR42J cells anddown-regulated miRNA-92b may decrease the apoptosis level of AR42J cells.