CAJ | 학술논문

目的：探讨高糖诱导心肌细胞损伤过程中氯离子浓度的变化以及氯离子通道阻断剂4,4’-二异硫氰基茋-2,2’-二磺酸（DIDS）对受损心肌细胞的作用。方法原代培养乳鼠心肌细胞，应用葡萄糖浓度为33mmol/L DMEM培养基，作用72h，构建心肌细胞损伤模型。实验分为对照组、高糖组、高糖+DIDS组和DIDS组。用MTT法检测细胞存活率，流式细胞仪检测细胞凋亡率，氯离子荧光探针（MQAE）检测细胞内氯离子浓度。结果高糖作用下的心肌细胞存活率呈浓度及时间依赖性降低（P＜0.05），流式细胞仪检测显示受损心肌细胞以凋亡性损伤为主。MQAE检测结果显示，与对照组比较，高糖组细胞内氯离子浓度显著下降（P＜0.05）。心肌细胞存活率及细胞内氯离子浓度，DIDS组与对照组比较差异均无统计学意义（P＞0.05）。与高糖组比较，高糖+DIDS组心肌细胞存活率显著升高，细胞内氯离子浓度下降明显减少，差异均有统计学意义（P＜0.05）。结论氯离子通道参与了高糖诱导的心肌细胞损伤过程，DIDS可在一定程度上减轻以凋亡为主的心肌细胞损伤。
목적：탐토고당유도심기세포손상과정중록리자농도적변화이급록리자통도조단제4,4’-이이류청기저-2,2’-이광산（DIDS）대수손심기세포적작용。방법원대배양유서심기세포，응용포도당농도위33mmol/L DMEM배양기，작용72h，구건심기세포손상모형。실험분위대조조、고당조、고당+DIDS조화DIDS조。용MTT법검측세포존활솔，류식세포의검측세포조망솔，록리자형광탐침（MQAE）검측세포내록리자농도。결과고당작용하적심기세포존활솔정농도급시간의뢰성강저（P＜0.05），류식세포의검측현시수손심기세포이조망성손상위주。MQAE검측결과현시，여대조조비교，고당조세포내록리자농도현저하강（P＜0.05）。심기세포존활솔급세포내록리자농도，DIDS조여대조조비교차이균무통계학의의（P＞0.05）。여고당조비교，고당+DIDS조심기세포존활솔현저승고，세포내록리자농도하강명현감소，차이균유통계학의의（P＜0.05）。결론록리자통도삼여료고당유도적심기세포손상과정，DIDS가재일정정도상감경이조망위주적심기세포손상。
Objective To investigate the change in chloride ion (Cl-) concentration in high glucose-induced cardiomyocyte injury and the role of 4,4’-diisothiocyanostilbene-2,2’-disulfonic acid (DIDS), a chloride channel blocker, in the injury. Methods Primarily cultured neonatal rat cardiomyocytes were exposed to glucose (33mmol/L) for 72h to establish the cell model of high glucose-induced injury. The cardiomyocytes were randomly divided into 4 groups:control group, high glucose group, high glucose+DIDS group and DIDS group. MTT assay was applied to detect cell viability, and flow cytometry was used to measure cell viability and apoptosis. N-(Ethoxycarbonylmethyl)-6-methoxyquinolinium bromide (MQAE) was used to examine the intracellular chloride concentration. Results High glucose decreased the cardiomyocyte viability in a dose- and time-dependent manner (P<0.05). Cardiomyocyte injury was mainly in the form of apoptosis. Compared to control group, high glucose resulted in a significant decrease in intracellular Cl-in the cardiomyocytes (P<0.05). There was no significant difference in cardiomyocyte viability and intracellular Cl-between the DIDS treated cells and control cells (P>0.05). While, DIDS treatment improved the cardiomyocyte viability and attenuated the decrease in intracellular Cl- in cardiomyocytes after high glucose induction (P<0.05). Conclusion Cl- is involved in the high glucose-induced cardiomyocyte injury. DIDS attenuates the injury which mainly presents as apoptosis.