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目的探讨番茄红素减轻H9C2心肌细胞缺氧/复氧(H/R)致内质网应激(ERS)的相关机制。方法 H9C2心肌细胞随机分为:(1)空白对照组、(2)番茄红素组、(3)H/R组、(4)番茄红素+H/R组、(5)4-苯基丁酸(4-phenyl butyric acid,4-PBA)+H/R组、(6)毒胡萝卜素内酯(thapsigargin,TG)组、(7)番茄红素+TG组。流式细胞术检测H9C2心肌细胞凋亡比例;Western blot检测葡萄糖调节蛋白(glucose-regulated proteins,GRP)78、C/EBP环磷酸腺苷反应元件结合转录因子同源蛋白(C/EBP homologous protein,CHOP)、c-Jun氨基末端激酶(c-Jun-N-terminal protein kinase,JNK)、磷酸化JNK(p-JNK)及半胱氨酸天冬氨酸蛋白酶(Casepase)-12的表达。结果与对照组相比,H/R组及TG组细胞的细胞凋亡比例、GRP78、CHOP、p-JNK和Caspase-12蛋白表达量明显增加(P<0.01);与H/R组相比,细胞凋亡比例及上述蛋白表达在番茄红素+H/R组与4-PBA+H/R组明显下降(P<0.01);与TG组相比,细胞凋亡比例及上述蛋白表达在番茄红素+TG组也明显下降(P<0.01);而番茄红素+H/R组与4-PBA+H/R组相比差异不具有统计学意义。JNK蛋白总量表达无明显变化。结论番茄红素通过抑制ERS致凋亡的相关信号通路CHOP、p-JNK和Caspase-12对H/R H9C2心肌细胞起到保护作用。
Objective To investigate the mechanism of lycopene in reducing endoplasmic reticulum stress (ERS) induced by hypoxia / reoxygenation (H / R) in H9C2 cardiomyocytes. Methods H9C2 cardiomyocytes were randomly divided into: (1) blank control group, (2) lycopene group, (3) H / R group, (4) lycopene + H / R group, 4-phenyl butyric acid (4-PBA) + H / R group, (6) thapsigargin group and (7) lycopene + TG group. Flow cytometry was used to detect the percentage of cardiomyocyte apoptosis in H9C2 cells. Western blot was used to detect the expression of glucose-regulated proteins (GRP), C / EBP homologous protein (C / EBP homologous protein, CHOP), c-Jun N-terminal protein kinase (JNK), phosphorylated JNK (p-JNK) and caspase-12. Results Compared with the control group, the percentage of apoptotic cells and the expression of GRP78, CHOP, p-JNK and Caspase-12 in H / R and TG groups were significantly increased (P <0.01). Compared with H / R group (P <0.01). The percentage of apoptosis and the expression of the above protein in lycopene + H / R group and 4-PBA + H / R group were significantly lower than those in TG group Lycopene + TG group also decreased significantly (P <0.01), but there was no significant difference between Lycopene + H / R group and 4-PBA + H / R group. The expression of JNK protein had no significant change. Conclusions Lycopene protects H / R H9C2 cardiomyocytes by inhibiting the signal pathways CHOP, p-JNK and Caspase-12 induced by ERS.