摘要
目的探讨转化生长因子-β1(TGF-β1)预处理调控缺氧诱导因子-1α(HIF-1α)的蛋白表达水平是否可以影响心肌细胞缺氧/复氧损伤。方法建立心肌细胞株H9C2缺氧/复氧损伤模型及缺氧预处理模型,将H9C2心肌细胞随机分为5组:正常对照组(Control组)、缺氧复氧组(H/R组)、缺氧预处理组(HPC组),TGF-β1预处理组(TGF-β1-Pre组)、TGF-β1抑制剂组(SB431542组),各组复氧结束后测定培养液乳酸脱氢酶(LDH)活性、HIF-1α蛋白表达水平、心肌细胞凋亡率。结果Control组、H/R组、HPC组、TGF-β1-Pre组、SB431542组的LDH活性(U/L)分别为(201.2±48.9)、(936.6±93.7)、(727.0±75.9)、(747.3±63.1)、(957.3±55.3);心肌细胞凋亡率(%)分别为(25.5±3.2)、(71.8±5.5)、(42.8±6.1)、(54.1±4.1)、(82.8±9.7),与H/R组比较,HPC组、TGF-β1-Pre组的LDH活性、心肌细胞凋亡率显著降低(P<0.05),HIF-1α蛋白表达水平升高(P<0.05);与HPC组比较,TGF-β1-Pre组各项指标差异均无统计学意义(P>0.05)。结论不论在常氧条件还是在缺氧环境下,TGF-β1均可以增加HIF-1α的蛋白表达水平,并且在缺氧复氧诱导所致的心肌细胞损伤中发挥抗凋亡作用,这种作用与调控HIF-1α的蛋白水平有关。
Objective To investigate whether the HIF-1α protein level regulated by TGF-β1 preconditioning can affect the cardiomyocytes hypoxia/reoxgenation-induced injury of cardiomyocytes. Methods H9C2 cardiomyocytes were used to establish hypoxia/reoxgenation-induced injury model. Cardiomyocytes were randomly divided into five groups: control group( Control),hypoxia/reoxgenation group( H/R),hypoxic preconditioning group( HPC),TGF-β1 preconditioning group( TGF-β1-Pre),TGF-β1 inhibitor group( SB431542). Lactic dehydrogenase( LDH) activity,HIF-1α protein content and cardiomyocytes apoptosis rate were measured at the end of the reperfusion. Results The LDH activity of Control group,H/R group,HPC group,TGF-β1-Pre group,SB431542 group were( 201.2±48.9) U/L,( 936.6±93.7) U/L,( 727.0±75.9) U/L,( 747.3± 63.1) U/L,( 957.3± 55.3) U/L,respectively.The cell apoptosis rate of Control group,H/R group,HPC group,TGF-β1-Pre group,SB431542 group were( 25.5±3.2) %,( 71.8±5.5) %,( 42.8±6.1) %,( 54.1±4.1) %,( 82.8± 9.7) %,respectively. The LDH activity and cell apoptosis rate in HPC group and TGF-β1-Pre group were both decreased compared with that in H/R group,meanwhile accompanied by increased HIF-1α protein content( P <0.05). There was no significantly difference between HPC group and TGF-β1-Pre group. Conclusion TGF-β1 could increase the expression level of HIF-1α protein no matter in normoxia or hypoxia circumstance,TGF-β1 exert antiapoptotic effect in hypoxia/reoxgenation-induced injury of cardiomyocytes,which is associated with the modulation of HIF-1α.
引文
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