| 佘正元,司晓明.微小 RNA-141-3p靶向调控含 CUE结构域蛋白 2基因对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响[J].安徽医药,2022,26(4):648-654. |
| 微小 RNA-141-3p靶向调控含 CUE结构域蛋白 2基因对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响 |
| Effect of microRNA-141-3p on the targeted regulation of the CUE domain-containing protein 2 gene on cerulein-induced injury of rat pancreatic acinar cells |
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| DOI：10.3969/j.issn.1009-6469.2022.04.003 |
| 中文关键词: 胰腺炎 细胞凋亡 bcl-2相关 X蛋白质 基因， bcl-2 肿瘤坏死因子 α 雨蛙素 微小 RNA-141-3p 含CUE结构域蛋白 2 |
| 英文关键词: Pancreatitis Apoptosis Bcl-2-related X protein Genes, bcl-2 Tumor necrosis factor-alpha Cerulein miR141-3p CUE domain containing protein 2 |
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| 中文摘要: |
| 目的探讨微小 RNA-141-3p(miR-141-3p)靶向调控含 CUE结构域蛋白 2(CUEDC2)对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响。方法 2018年7月至 2019年12月，从美国 ATCC购买大鼠胰腺腺泡细胞 AR42J。100 nm/L雨蛙素(CAE)刺激大鼠胰腺腺泡细胞 AR42J 6 h建立急性胰腺炎细胞模型。实时荧光定量 PCR(qRT-PCR)和蛋白质印迹法(Western blotting)检测雨蛙素干预后AR42J细胞中 miR-141-3p和CUEDC2的表达水平。双荧光素酶报告基因实验和蛋白质印迹法验证 miR-141-3p和CUEDC2的靶向关系。利用脂质体转染法将 miR-141-3p抑制物(anti-miR-141-3p)、miRNA抑制物阴性对照(anti-miR-NC)、CUEDC2过表达载体(pcDNA-CUEDC2)、空载体(pcDNA)分别转染 AR42J细胞，经雨蛙素干预处理后，流式细胞术检测细胞凋亡，蛋白质印迹法检测 B细胞淋巴瘤 /白血病 -2(Bcl-2)和Bcl-2相关 X蛋白(Bax)的表达水平，酶联免疫吸附试验(Elisa)试剂盒检测细胞培养上清中肿瘤坏死因子-α(TNF-α)和白细胞介素 -6(IL-6)水平。结果雨蛙素干预处理后 AR42J细胞中 miR-141-3p的表达水平显著升高［(2.43±0.24)比(1.00±0.09)］，CUEDC2的表达水平显著降低。 miR-141-3p靶向负性调控 CUEDC2表达。与转 anti-miR-NC和雨蛙素协同处理组比较，转染 anti-miR-141-3p和雨蛙素协同处理组 AR42J细胞凋亡率显著增加［(27.48±2.33)%比(15.64±1.51)%］Bax蛋白的表达显著增加， Bcl-2蛋白的表达显著降低， TNF-α［(136.54±14.58)ng/L比( 226.48±20.54)ng/L］和 IL-6［(115.89±11.65)n，g/L比( 193.47± 18.63)ng/L］的分泌显著降低；与转染 pcDNA和雨蛙素协同处理组比较，转染 pcDNA-CUEDC2和雨蛙素协同处理组 AR42J细胞凋亡率显著增加［(23.87±2.35)%比(14.23±1.44)%］，Bax蛋白的表达显著增加， Bcl-2蛋白的表达显著降低， TNF-α［(158.74±15.32)ng/L(236.87±18.66)ng/L］和IL-6［(135.77±14.97)ng/L比(189.67±17.32)ng/L］的分泌显著降低；转染 si-CUEDC2可逆转转染 anti-miR14比1-3p对雨蛙素诱导的大鼠胰腺腺泡细胞损伤的影响。结论抑制 miR-141-3p通过靶向 CUEDC2可促进急性胰腺炎腺泡细胞的凋亡，抑制 TNF-α和IL-6分泌，从而减轻雨蛙素诱导的大鼠胰腺腺泡细胞损伤。 |
| 英文摘要: |
| Objective To investigate the effect of microRNA-141-3p (miR-141-3p) targeting CUE domain containing 2 protein (CUEDC2) on cerulein-induced pancreatic acinar cell injury in rats.Methods This study was conducted from July 2018 to December2019. Rat AR42J pancreatic acinar cells were purchased from ATCC (USA). A cell model of acute pancreatitis was established by stim ulating rat pancreatic acinar AR42J cells with 100 nm/L cerulein (CAE) for 6 h. Real-time quantitative PCR (qRT-PCR) and Western blotting were used to detect the expression levels of miR-141-3p and CUEDC2 in AR42J cells after cerulein intervention. Dual-lucifer ase reporter gene assay and Western blotting verified the targeting relationship between miR-141-3p and CUEDC2. miR-141-3p inhibi tor (anti-miR-141-3p), miRNA inhibitor negative control (anti-miR-NC), CUEDC2 overexpression vector (pcDNA-CUEDC2) and emptyvector (pcDNA) were transfected into AR42J cells by lipofection, and after treatment with cerulein, cell apoptosis was detected by flowcytometry, and the expression of B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2 related X protein (Bax) were detected by Western blot ting. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the cell culture supernatant were detected by an enzyme-linked immunosorbent assay (ELISA) kit.Results After treatment with cerulein, the expression level of miR-141-3p in AR42J cells was significantly increased [(2.43±0.24) vs. (1.00±0.09)], and the expression level of CUEDC2 was significantly decreased. miR-141-3p targets and negatively regulates CUEDC2 expression. Compared with the transfected anti-miR-NC and cerulein cotreatment group, the AR42J cell apoptosis rate significantly increased in the transfected anti-miR-141-3p and cerulein cotreatment group [(27.48±2.33)% vs. |
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