钙敏感受体(CaSR)在高糖诱导H9C2细胞氧化应激中的作用

Role of calcium-sensing receptor (CaSR) in high glucose-induced oxidative stress of H9C2 cells

目的 本研究旨在观察钙敏感受体（calcium-sensing receptor,CaSR）在高糖诱导H9C2心肌细胞中的表达以及其对氧化应激的作用.方法 将H9C2细胞在不同条件下干预24 h,包括正常糖浓度组（NG组）、高糖组（HG组）、高糖＋CaSR激动剂组（HG＋GdCl3组）、高糖＋CaSR抑制剂组（HG＋NPS2390组）.用CCK8试剂盒检测细胞存活率;DCFH-DA荧光探针检测细胞内活性氧簇（ROS）含量;用比色法检测细胞超氧化物歧化酶（SOD）和谷胱甘肽过氧化物酶（GSH-Px）活性以及丙二醛（MDA）含量.用Western blot检测细胞CaSR蛋白表达水平.结果 细胞存活率呈糖浓度依赖性降低[（89.00±1.54）％比（98.83±0.47）％,P＜0.01],CaSR蛋白表达量呈糖浓度依赖性增加2.14±0.09比1.06±0.05,P＜0.01）.高糖诱导可使细胞发生氧化应激,SOD（14.14±0.57比22.86±0.58,P＜0.01）与GSH-Px（2.62±0.59比3.04±0.89,P＜0.01）活性降低,ROS（0.040±0.002比0.020±0.001,P＜0.01）与MDA（0.83±0.05比0.54±0.02,P＜0.01）生成增多.与HG组相比,HG＋GdCl3组细胞氧化损伤加重（P＜0.05）.而HG＋NPS2390组细胞氧化损伤减轻（P＜0.05）.结论 高糖能上调CaSR蛋白表达进而促进氧化应激的发生.

Objective To observe the protein expression of calcium-sensing receptor （CaSR） and the effect of CaSR on high glucose induced-oxidative stress in H9C2 cells. Methods H9C2 cells were exposed to different treatments for 24 h, including normal glucose group（NG group）, high glucose group（HG group）, HG＋CaSR activator group （HG＋GdCh group）, HG＋CaSR inhibitor group （HG＋NPS2390 group ）. Cell viability was detected by CCK8 kit. Reactive oxygen species （ROS） generation was detected by fluorescent probe DCFH-DA. The activi- ty of superoxide dismutase （SOD） and glutathion peroxidase （GSH-Px） , and the content of malonaldehyde （MDA） were measured by colorimetry. In addition, the expression level of CaSR was determined by Western blot. Results The cell viability decreased [ （89.00± 1.54）% vs. （98.83±0.47）%, P〈0.01 ] and the CaSR protein level increased （2.14±0.09 vs. 1.06±0.05, P〈0.01 ） in a glucose-concentration dependent manner. High glucose exposure induced cell oxidative stress, decreased the activity of SOD （14.14±0.57 vs. 22.86±0.58, P〈0.01 ） and GSH-Px （2.62±0.59 vs. 3.04±0.89 ,P〈0.01 ）, increased ROS （0.040±0.002 vs. 0.020±0.001 ,P〈0.01 ） and MDA（0.83±0.05 vs. 0.54±0.02, P〈0.01 ） content. Compare with HG group, the cell oxidative stress damage in HG＋GdC13 group was significantly aggravated （P〈0.05）, while that in HG＋NPS2390 group was significantly alleviated （P〈0.05）. Conclusion High glucose up-regulated the protein expression of CaSR, following enhanced the oxidative stress.