鞠加圣,陈建良,杨云峰,彭智涛,钟远强,杨 淬.miR-1298表达对PC-12细胞糖氧剥夺/复氧损伤的影响及其机制研究[J].,2022,(12):2212-2216 |
miR-1298表达对PC-12细胞糖氧剥夺/复氧损伤的影响及其机制研究 |
Effect of miR-1298 Expression on Oxygen Glucose Deprivation/Reoxygenation Injury of PC-12 Cells and its Mechanism Study |
投稿时间:2022-01-06 修订日期:2022-01-28 |
DOI:10.13241/j.cnki.pmb.2022.12.003 |
中文关键词: miR-1298 PC-12细胞 糖氧剥夺/复氧 凋亡 BCL-2 BAX |
英文关键词: miR-1298 PC-12 cells Oxygen glucose deprivation/reoxygenation Apoptosis BCL-2 BAX |
基金项目:国家自然科学基金地区科学基金项目(81760655) |
|
摘要点击次数: 719 |
全文下载次数: 413 |
中文摘要: |
摘要 目的:通过体外细胞培养探讨miR-1298对缺血缺氧性神经损伤的调节作用。方法:首先通过细胞活性检测和乳酸脱氢酶(LDH)细胞毒性法确定大鼠PC-12细胞糖氧剥夺/复氧(OGD/R)的造模效果,同时采用实时荧光定量PCR(RT-qPCR)检测细胞miR-1298的表达差异。体外转染miR-1298mimic、mimic NC、miR-1298 inhibitor和inhibitor NC至大鼠PC-12细胞系,检测mimic、mimic NC、inhibitor、inhibitor NC的转染效率。经过OGD/R处理后将细胞分为Control组、OGD/R组、mimic组、mimicNC组、inhibitor组和inhibitorNC组。流式细胞术检测各组PC-12细胞凋亡的情况,免疫印迹试验(Western blot)检测各组PC-12细胞凋亡相关蛋白B淋巴细胞瘤-2基因(BCL-2)和Bcl-2相关的x基因(Bax)表达的情况。结果:PC12细胞经过OGD/R处理后,其细胞存活率与Control组比明显下降且LDH漏出率明显上升(均P<0.05);模型细胞中miR-1298相对表达量明显低于Control组(P<0.05)。转染24小时后mimic组细胞中miR-1298的相对表达量明显高于mimicNC组(P<0.05);mimic组细胞凋亡率低于mimicNC组,而inhibitor组细胞凋亡率高于inhibitor NC组(均P<0.05);mimic组的BCL-2表达量较mimicNC组升高,而BAX表达量下降,inhibitor组与inhibitorNC组相比,BCL-2表达量下降,而BAX表达量上升,差异均有统计学意义(均P<0.05)。结论:miR-1298通过抑制细胞凋亡减轻PC-12细胞OGD/R的损伤。 |
英文摘要: |
ABSTRACT Objective: To investigate the regulatory effect of miR-1298 on hypoxic-ischemic nerve injury by cell culture in vitro. Methods: Firstly, the modeling effect of oxygen glucose deprivation/reoxygenation (OGD/R) of rat PC-12 cells was determined by cell activity detection and lactate dehydrogenase (LDH) cytotoxicity. At the same time, the expression difference of miR-1298 was detected by real-time fluorescence quantitative PCR (RT-qPCR). The miR-1298mimic, mimic NC, miR-1298 inhibitor and inhibitor NC were transfected into rat PC-12 cell line in vitro. The transfection efficiency of miR, mimic NC, inhibitor and inhibitor NC was detected. After OGD/R treatment, the cells were divided into control group, OGD/R group, mimicgroup, mimicNC group, inhibitor group and inhibitor NC group. The apoptosis of PC-12 cells in each group was detected by flow cytometry, and the expression of PC-12 apoptosis related protein B lymphoma-2 gene (BCL-2) and Bcl-2 associated X protein (BAX) were detected by Western blot. Results: After OGD/R treatment, the survival rate of PC12 cells decreased significantly compared with the control group, and the leakage rate of LDH increased significantly (all P<0.05). The relative expression of miR-1298 in model cells was significantly lower than that in control group (P<0.05). 24 hours after transfection, the relative expression of miR-1298 in mimic group cells was significantly higher than that in mimicNC group (P<0.05). The cells apoptosis rate in mimic group was lower than that in mimicNC group, while cells apoptosis rate in inhibitor group was higher than that in inhibitor NC group (all P<0.05). The expression of BCL-2 in mimic group was higher than that in mimicNC group, while the expression of BAX decreased. Compared with the inhibitorNC group, the expression of BCL-2 decreased, while the expression of BAX increased, and the differences were statistically significant (all P<0.05). Conclusion: miR-1298 alleviates the injury of OGD/R in PC-12 cells by inhibiting apoptosis. |
查看全文
查看/发表评论 下载PDF阅读器 |
关闭 |
|
|
|