CAJ | 학술논문

目的：研究ATP敏感性钾通道（ KATP通道）在硫化氢（ H2 S）抑制高糖引起心肌损伤中的作用。方法：应用Western blot 法检测心肌细胞KATP通道蛋白的表达水平；CCK-8试剂盒测定心肌细胞存活率；Hoechst 33258染色测定凋亡细胞数量的变化；JC-1染色法测定线粒体膜电位（ MMP ）。结果：应用高糖（35 mmol／L葡萄糖）处理H9c2细胞1～24 h，其中6 h、9 h、12 h和24 h均能明显下调KATP通道蛋白的水平，12 h和24 h KATP水平降至最低。在HG处理心肌细胞12 h前，应用400μmol／L硫氢化钠（ NaHS，为H2 S的供体）预处理30 min明显抑制高糖对KATP通道蛋白表达的下调作用。100μmol／L线粒体KATP通道开放剂二氮嗪和50μmol／L非选择性KATP通道开放剂吡拉地尔（ Pin）及NaHS预处理均显著抑制高糖引起的心肌细胞损伤，使细胞存活率升高，凋亡细胞数量及MMP丢失减少。相反，100μmol／L线粒体KATP通道阻断剂5-羟基癸酸和1 mmol／L非选择性KATP通道阻断剂格列本脲均能明显阻断上述NaHS的心肌细胞保护作用。结论：KATP通道介导了H2 S对高糖引起的心肌细胞损伤的抑制作用。
목적：연구ATP민감성갑통도（ KATP통도）재류화경（ H2 S）억제고당인기심기손상중적작용。방법：응용Western blot 법검측심기세포KATP통도단백적표체수평；CCK-8시제합측정심기세포존활솔；Hoechst 33258염색측정조망세포수량적변화；JC-1염색법측정선립체막전위（ MMP ）。결과：응용고당（35 mmol／L포도당）처리H9c2세포1～24 h，기중6 h、9 h、12 h화24 h균능명현하조KATP통도단백적수평，12 h화24 h KATP수평강지최저。재HG처리심기세포12 h전，응용400μmol／L류경화납（ NaHS，위H2 S적공체）예처리30 min명현억제고당대KATP통도단백표체적하조작용。100μmol／L선립체KATP통도개방제이담진화50μmol／L비선택성KATP통도개방제필랍지이（ Pin）급NaHS예처리균현저억제고당인기적심기세포손상，사세포존활솔승고，조망세포수량급MMP주실감소。상반，100μmol／L선립체KATP통도조단제5-간기계산화1 mmol／L비선택성KATP통도조단제격렬본뇨균능명현조단상술NaHS적심기세포보호작용。결론：KATP통도개도료H2 S대고당인기적심기세포손상적억제작용。
AIM:To investigate the roles of ATP-sensitive potassium ( KATP ) channels in high glucose-induced cardiac injury and the inhibitory effect of hydrogen sulfide ( H2 S) on the cardiomyocyte injury.METHODS:The expres-sion level of KATP channel protein was tested by Western blot.The cell viability was measured by CCK-8 assay.The number of apoptotic cells was observed by Hoechst 33258 nuclear staining.Mitochondrial membrane potential ( MMP) was exam-ined by JC-1 staining.RESULTS:After the H9c2 cells were treated with 35 mmol/L glucose ( high glucose, HG) for 1~24 h, the protein level of KATP channel was significantly reduced at 6 h, 9 h, 12 h and 24 h, reaching the minimum level at 12 h and 24 h.Pretreatment of the cells with 400μmol/L NaHS ( a donor of H2 S) prior to exposure to HG for 12 h con-siderably blocked the down-regulation of KATP channels induced by HG.Pretreatment of the cells with 100 μmol/L mito-chondrial KATP channel opener diazoxide, 50μmol/L non-selective KATP channel opener pinacidil or NaHS obviously inhibi-ted HG-induced injuries, leading to an increase in the cell viability, and decreases in the number of apoptotic cells and the MMP loss.Pretreatment with 100μmol/L mitochondrial KATP channel antagonist 5-hydroxydecanoic acid or 1 mmol/L non-selective KATP channel antagonist glibenclamide attenuated the above cardioprotective effects of NaHS.CONCLUSION:KATP channels mediate the inhibitory effect of H2 S on HG-induced cardiac injury.