黑果枸杞花青素对低氧诱导的H9c2大鼠心肌细胞凋亡的影响

doi:10.11843/j.issn.0366-6964.2023.08.033

摘要/Abstract

摘要： 旨在探究青海柴达木黑果枸杞（Lycium ruthenicum Murray）花青素（anthocyanidin，AC）对低氧诱导的H9c2大鼠心肌细胞凋亡的影响，作者选用H9c2大鼠心肌细胞为研究对象，设常氧组、低氧组、低氧+黑果枸杞花青素组和低氧+红景天苷（salidroside，Sal）组，采用酶标仪检测其细胞乳酸脱氢酶（lactic dehydrogenase，LDH）含量、显微镜观察其细胞形态学变化、荧光酶标仪检测其细胞内活性氧（reactive oxygen species，ROS）含量、Hoechst33342/PI双染法检测其细胞凋亡程度、流式细胞术检测其细胞凋亡率、qRT-PCR法和Western blot法分别检测细胞凋亡因子B淋巴细胞瘤-2（B-cell lymphoma-2，Bcl2）及Bcl2相关蛋白（Bcl2-associated X，Bax）在mRNA水平和蛋白水平的表达。结果表明，与常氧组相比，低氧能极显著增加LDH、ROS含量和细胞凋亡率（P<0.01），极显著下调Bcl2、Bcl-2/Bax比值的mRNA和蛋白表达（P<0.01），极显著上调Bax的mRNA和蛋白表达（P<0.01）。在低氧条件下，黑果枸杞花青素可显著减少LDH、ROS含量（P<0.05），细胞凋亡率极显著减少（P<0.01），显著上调Bcl-2和Bcl2/Bax比值的mRNA和蛋白表达（P<0.05），极显著下调Bax的蛋白表达（P<0.01）。综上可知，在低氧条件下，黑果枸杞花青素可通过降低LDH和ROS含量，上调Bcl2和Bcl2/Bax比值在mRNA水平和蛋白水平的表达，下调Bax的蛋白表达来有效延缓低氧诱导的H9c2大鼠心肌细胞的凋亡，对低氧诱导的H9c2大鼠心肌细胞具有保护作用。

关键词: 黑果枸杞, 花青素, H9c2大鼠心肌细胞, 低氧, 凋亡

Abstract: To investigate the effect of anthocyanins from Lycium ruthenicum Murray of Qaidam, Qinghai, on hypoxia-induced apoptosis of H9c2 rat cardiomyocytes, the H9c2 rat cardiomyocytes were selected for the study, including normoxia group, hypoxia group, hypoxia+anthocyanin group and hypoxia+salidroside group, LDH content was detected by enzyme label, cell morphological changes were observed by microscope, the content of ROS was explored by fluorescent enzyme labeling instrument, cell apoptosis was detected by Hoechst33342/PI double staining and flow cytometry. In addition, the expression of apoptosis related factors (Bcl2, Bax) at mRNA and protein levels were tested by qRT-PCR and Western blot, respectively. The results showed that compared with the normoxia group, hypoxia significantly increased the contents of LDH, ROS and the rate of apoptosis (P<0.01), and down-regulated the expression of Bcl2 and Bcl-2/Bax (P<0.01), the mRNA and protein expression of Bax were significantly up-regulated (P<0.01). Under hypoxic condition, anthocyanin significantly reduced the contents of LDH and ROS (P<0.05), significantly reduced the rate of apoptosis (P<0.01), significantly up-regulated the expression of Bcl2 and Bcl2/Bax ratio mRNA and protein (P<0.05), the protein expression of Bax was significantly down-regulated (P<0.01). In conclusion, under hypoxic conditions, Lycium ruthenicum Murray anthocyanin can effectively delay the apoptosis of H9c2 rat cardiomyocytes induced by hypoxia by reducing the content of LDH and ROS in cells, up regulating the expression of Bcl2 and Bcl2/Bax ratio at mRNA level and protein level, and down regulating the expression of Bax at protein level.

Key words: Lycium ruthenicum Murray, anthocyanidin, H9c2 rat cardiomyocytes, hypoxia, apoptosis

中图分类号:

S852.3

沈童, 王梦杰, 吴华, 李金铭, 王硕, 武小庆, 陈鑫磊, 行倩文, 刘波. 黑果枸杞花青素对低氧诱导的H9c2大鼠心肌细胞凋亡的影响[J]. 畜牧兽医学报, 2023, 54(8): 3490-3499.

SHEN Tong, WANG Mengjie, WU Hua, LI Jinming, WANG Shuo, WU Xiaoqing, CHEN Xinlei, XING Qianwen, LIU Bo. Effect of Lycium ruthenicum Murray Anthocyanin on Hypoxia-induced Apoptosis in H9c2 Rat Cardiomyocytes[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3490-3499.

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