| 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） |
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| 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. |
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