التفاصيل البيبلوغرافية
| العنوان: |
旋毛虫 HSP70 蛋白诱导大鼠心肌细胞线粒体凋亡机制的研究. (Chinese) |
| العنوان البديل: |
Effects of Trichinella spiralis HSP70 protein on apoptosis related protein expression in rat cardiomyocytes. (English) |
| المؤلفون: |
王姗姗, 刁子洋, 郭 琨, 王雪莹, 侯嘉明, 薄禄琪, 王 爽, 宋铭忻 |
| المصدر: |
Chinese Journal of Preventive Veterinary Medicine / Zhongguo Yufang Shouyi Xuebao; Aug2022, Vol. 44 Issue 8, p881-887, 7p |
| مصطلحات موضوعية: |
TRICHINELLA spiralis, RECOMBINANT proteins, GENE expression, LACTATE dehydrogenase, HEART injuries, APOPTOSIS, HEART cells |
| الملخص (بالإنجليزية): |
To investigate the mechanism of cardiac injury induced by Trichinella spiralis, the recombinant protein of Trichinella spiralis HSP70 (Ts-HSP70) was applied to rat H9c2 myocardial cells. The optimum concentration of Ts-Hsp70 stimulated H9c2 cells was dertermined by CCK-8 kit. The amount of lactate dehydrogenase (LDH) was determined by microplate method, and methodcreatine kinase (CK) as marker of myocardial injury was measured by colorimetry. ROS release was detected by immunofluorescence as well as western blot. qPCR were used to detect the expression of Bax, Caspase-3, Caspase-9, Cytochrome C, and Bcl-2 in mitochondrial apoptosis and anti- apoptosis pathways. The results showed that when the optimal concentration of Ts-HSP70 at 60μg/mL stimulated H9c2 cells for 6 hours, 12 hours and 18 hours, with the extension of the culturing time of Ts- HSP70 on H9c2 cells, the amount of LDH, CK, ROS and the gene transcription and protein expression level of Bax, Cytochrome C and activated Caspase-3, Caspase-9 gene were significantly higher than that of control group (P<0.01), and reached the peak at 12 hours. Meanwhile, the expression of mitochondria antiapoptotic gene Bcl- 2 was significantly lower than control group (P<0.01), and reached the lowest at 12 hours. The results for the first time showed that Ts-Hsp70 could increase the amount of LDH, CK and ROS in H9c2 cells, and activate and regulate the mitochondrial apoptosis pathway in H9c2 cells. The results provided basic infromation for further research on the mechanism of cardiac injury induced by Trichinella spiralis. [ABSTRACT FROM AUTHOR] |
| Abstract (Chinese): |
为了解旋毛虫 HSP70 致心肌损伤的机制，本研究将不同浓度旋毛虫重组 HSP70 蛋白（Ts-Hsp70）与 大鼠心肌细胞 H9c2 共孵育后，采用 CCK-8 试剂盒筛选 Ts-Hsp70 作用 H9c2 细胞的最适浓度。以最适浓度 Ts- Hsp70 与 H9c2 细胞共孵育不同时间后，采用微孔板法检测细胞培养上清液中乳酸脱氢酶（LDH）的含量，采用比 色法检测细胞培养上清液中心肌细胞损伤标志物肌酸激酶（CK）的含量，采用免疫荧光检测 Ts-Hsp70 作用后 H9c2 细胞活性氧（ROS）的释放，采用 qPCR 和 western blot 方法检测 Ts-Hsp70 作用后的 H9c2 细胞线粒体凋亡与抗凋亡 途径中的 Bax、Caspase-3、Caspase-9、Cytochrome C 与 Bcl-2 的基因转录水平与蛋白表达水平。结果显示，经最 适浓度为 60 μg/mL 的 Ts-Hsp70 分别刺激 H9c2 细胞 6 h、12 h 和 18 h 后，随着 Ts-Hsp70 对 H9c2 细胞作用时间的延 长，LDH、CK、ROS 含量与 Bax、Cytochrome C 及激活状态下的 Caspase-3、Caspase-9 基因转录水平与蛋白表达水 均升高，且均于作用 12 h 时达峰值，显著高于对照组（P<0.01），而线粒体抗凋亡基因 Bcl-2 基因转录水平与蛋白 表达水平随着 Ts-Hsp70 对 H9c2 细胞作用时间的延长均逐渐降低，且均于作用 12 h 时达谷值，显著低于对照组 （P<0.01）。本研究首次证实 Ts-Hsp70 可增加细胞中 LDH、CK、ROS 的含量，激活并调节 H9c2 细胞中的线粒体凋 亡通路，该研究结果为进一步研究旋毛虫致心肌损伤机制提供了基础数据。 [ABSTRACT FROM AUTHOR] |
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| قاعدة البيانات: |
Complementary Index |
