左旋卡尼汀联合CrkL降低缺氧复氧诱导的心肌细胞损伤

目的研究左旋卡尼汀联合CT10激酶调节子样蛋白(CrkL)降低缺氧复氧(H/R)诱导的心肌细胞损伤的作用。方法利用H/R损伤心肌细胞H9c2,使用左旋卡尼汀处理。细胞计数试剂盒8(CCK-8)、流式细胞术、Western blot分别检测细胞的增殖、凋亡和细胞核相关抗原Ki-67(Ki-67)、增殖细胞核抗原(PCNA)、B细胞淋巴瘤/白血病2(Bcl-2)、Bcl-2相关X蛋白(Bax)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、核因子κB(NF-κB)、CrkL水平。在细胞H9c2中转染pcDNA-CrkL,使用H/R处理或H/R+左旋卡尼汀处理。采用上述方法检测细胞增殖、凋亡等。结果与对照组比较,H/R组H9c2细胞的活力、Ki-67、PCNA、Bcl-2、CrkL蛋白表达量明显降低,细胞凋亡率、Bax蛋白水平及炎症因子TNF-α、IL-1β、NF-κB的蛋白水平显著升高(P<0.05)。与H/R组比较,左旋卡尼汀明显增加H/R诱导的H9c2细胞活力及Ki-67、PCNA、Bcl-2、CrkL蛋白表达量,显著降低细胞凋亡率、Bax蛋白水平及TNF-α、IL-1β、NF-κB蛋白水平(P<0.05)。CrkL过表达明显提高H/R诱导的H9c2细胞活力和Ki-67、PCNA、Bcl-2蛋白表达量,显著降低细胞凋亡率、Bax蛋白水平及TNF-α、IL-1β、NF-κB蛋白水平(P<0.05)。与单独使用左旋卡尼汀或CrkL过表达比较,左旋卡尼汀联合CrkL过表达明显提高H9c2细胞的活力和Ki-67、PCNA、Bcl-2蛋白表达量,显著降低细胞凋亡率、Bax蛋白水平及TNF-α、IL-1β、NF-κB蛋白水平(P<0.05)。结论左旋卡尼汀联合CrkL可以促进缺氧复氧诱导的心肌细胞增殖,降低细胞凋亡和炎症反应,从而保护心肌细胞。

L-carnitine combined with CrkL reduces hypoxia/reoxygenation-induced cardiomyocyte injury

Aim To study the effect of L-carnitine combined with CT10 regulator of kinase like protein(CrkL)on reducing myocardial cell injury induced by hypoxia/reoxygenation(H/R).Methods H9c2 cells were injured by H/R and treated with L-carnitine.CCK-8,flow cytometry,and Western blot were applied to determine cell proliferation,apoptosis,and nuclear associated antigen Ki67(Ki-67),proliferating cell nuclear antigen(PCNA),B cell lymphoma/lewkmia-2(Bcl-2),Bcl-2 associated X protein(Bax),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),n uclear factor-κB(NF-κB),and CrkL levels,respectively.Cells H9c2 were transfected with pcDNA-CrkL,and treated with H/R or treated with H/R and L-carritine.The above methods were used to detect cell proliferation and apoptosis.Results Compared with the control group,the viability,Ki-67,PCNA,Bcl-2,and CrkL protein expression of H9c2 cells were significantly decreased in the H/R group,and the apoptosis rate,Bax protein level,and the levels of inflammatory factors TNF-α,IL-1β,NF-κB were evidently increased(P<0.05).Compared with the H/R group,L-carnitine obviously improved the H/R-induced H9c2 cell viability,Ki-67,PCNA,Bcl-2,and CrkL protein expression,and remarkably reduced the apoptosis rate,Bax protein level,and TNF-α,IL-1β,NF-κB levels(P<0.05).CrkL overexpression dramatically enhanced H/R-induced H9c2 cell viability,Ki-67,PCNA,and Bcl-2 protein expression,while markedly reduced apoptosis rates,Bax protein levels,TNF-α,IL-1β,and NF-κB levels(P<0.05).Compared with L-carnitine or CrkL overexpression alone,L-carnitine combined with CrkL overexpression clearly increased the viability of H9c2 cells,Ki-67,PCNA,and Bcl-2 protein expression,and distinctly reduced the apoptosis rate and Bax protein level,TNF-α,IL-1β,NF-κB levels(P<0.05).Conclusion L-carnitine combined with CrkL promotes the proliferation of cardiomyocytes induced by H/R,reduces apoptosis and inflammatory response,and thus protects cardiomyocytes from damage.