苦参碱对高糖诱导的H9C2心肌细胞损伤的影响

Effect of matrine on high glucose-induced injury of H9C2 cardiomyocytes

目的探讨苦参碱对高糖诱导的H9C2心肌细胞损伤的影响。方法将H9C2心肌细胞随机分为正常糖组、正常糖苦参碱组、高糖组和高糖苦参碱组。实时荧光定量PCR检测超氧化物歧化酶2(SOD2)、Gp91phox、BAX、BCL-2、TNF-α、白细胞介素(IL)-1β和IL-6 mRNA表达;Western blot检测SOD2、P67phox、BAX、BCL-2、剪切形式半胱氨酸天冬氨酸蛋白酶3(C-Caspase-3)、NF-κB P65亚基磷酸化(P-P65)和NF-κB P65总蛋白(T-P65);试剂盒检测细胞内丙二醛质量、谷胱甘肽过氧化物酶(GSH-Px)及过氧化氢酶(CAT)活性;TUNEL检测细胞凋亡。结果正常糖组与正常糖苦参碱组丙二醛和GSH-Px及CAT活性等指标比较,无统计学差异(P>0.05);与正常糖组比较,高糖组丙二醛、P67phox、P-P65、BAX、C-Caspase-3蛋白、Gp91phox、IL-1β、IL-6、TNF-αmRNA表达和细胞凋亡率明显升高,GSH-Px、CAT活性明显降低[(0.98±0.02)U/mg vs(3.82±0.05)U/mg,(12.18±0.52)U/mg vs(34.82±0.63)U/mg,P<0.05],SOD2、BCL-2蛋白及mRNA表达明显下调(P<0.05);与高糖组比较,高糖苦参碱组丙二醛、P67phox、P-P65、BAX、C-Caspase-3蛋白、Gp91phox、IL-1β、IL-6、TNF-αmRNA表达和细胞凋亡率明显降低,GSH-Px、CAT活性、SOD2、BCL-2蛋白及mRNA表达明显上调(P<0.05)。结论苦参碱可以明显改善高糖诱导的H9C2心肌细胞损伤,这可能与其抗氧化、抗炎和抗凋亡活性有关。

Objective To study the effect of matrine on high glucose-induced injury of H9C2 cardiomyocytes.Methods H9C2 cardiomyocytes were randomly divided into normal glucose group,normal glucose+matrine group,high glucose group and high glucose+matrine group.The expressions of SOD2,Gp91phox,BAX,BCL-2,TNF-α,IL-1βand IL-6 mRNA were detected by quantitative RT-PCR and those of SOD2,P67phox,BAX,BCL-2,C-Caspase 3,P-P65,T-P65 proteins were detected by Western blot.The MDA content,GSH-Px and CAT activities were assayed with specific kits and the cell apoptosis was assessed with TUNEL staining.Results No significant difference was detected in cell apoptosis,MDA content,GSH-Px and CAT activities between normal glucose group and normal glucose+matrine group(P>0.05).The expression levels of MDA,P67phox,P-P65,BAX,C-Caspase 3 proteins and Gp91phox,IL-1β,IL-6,TNF-αmRNA and the apoptotic rate of H9C2 cardiomyocytes were significantly higher while the activities of GSH-Px and CAT(0.98±0.02 U/mg vs 3.82±0.05 U/mg,12.18±0.52 U/mg vs 34.82±0.63 U/mg,P<0.05)and the expression levels of SOD2 and BCL-2 protein and mRNA were significantly lower in high glucose group than in normal glucose group,which were notably reversed by matrine treatment(P<0.05).Conclusion Matrine can significantly improve high glucose-induced injury of H9C2 cardiomyocytes due to its anti-oxidative,anti-inflammatory and anti-apoptotic effects.