黄芪总黄酮通过调控miR-190a-5p对缺氧/复氧诱导的心肌细胞损伤的影响
wf(2022)
Abstract
目的:探讨黄芪总黄酮(TFA)对缺氧/复氧(H/R)诱导心肌细胞损伤的影响及其可能作用机制。方法:空白培养H9c2细胞记为对照组,H9c2细胞放入缺氧培养箱内(37℃、95%N2、5%CO2) 4 h,更换培养基,放入正常培养箱内复氧处理12 h,记为H/R组;50、100、150 μg/mL TFA培养H9c2细胞24 h后进行H/R处理,记为H/R+TFA-L组、H/ R+TFA-M组、H/R+TFA-H组。miR-NC、miR-190a-5p mimics分别转染至H9c2细胞后进行H/ R处理,记为H/R+miR-NC组、H/R+miR-190a-5p组;anti-miR-NC、anti-miR-190a- 5p分别转染至H9c2细胞后用150 μg/mL TFA培养H9c2细胞24 h,并进行H/R处理,记为H/R+ TFA-H+anti-miR-NC组、H/R+TFA-H+anti-miR-190a-5p组。RT-PCR检测miR-190a-5p表达量;采用流式细胞术检测细胞凋亡率;试剂盒检测SOD、MDA、LDH和ROS水平;Western blot检测cleaved caspase-3、BCL2蛋白表达量。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析。结果:与对照比较,H/R组细胞凋亡率[(32.76±2.42)%比(3.23±0.43)%]、cleaved caspase-3 (0.83±0.05比0.26±0.02)、MDA[(23.36±2.25)比(4.26±0.48)nmol/mg]、LDH [(52.51±3.82)比(13.66±1.72)U/L]、ROS[(192.63±10.42)%比(98.51±6.55)%]水平增高(P均< 0.05),BCL2蛋白水平(0.33±0.06比0.91±0.05)、SOD [(52.14±4.95)比(135.32±9.22)U/mg]水平、miR-190a-5p表达量(0.32±0.05比1.02±0.10)降低(P均< 0.05);与H/R组比较,H/R+TFA-L组、H/R+TFA-M组、H/R+TFA-H组细胞凋亡率[(27.09±2.11)%、(19.21±1.59)%、(11.01±1.25)%比(32.76±2.42)%]、cleaved caspase-3、MDA [(18.17± 1.67)、(14.08±1.22)、(9.84±0.73)比(23.36±2.25)nmol/mg]、LDH [(43.11±4.30)、(31.25± 2.59)、(19.42±1.89)比(52.51±3.82) U/L]、ROS水平[(166.11±9.88)%、(141.52±8.94)%、(115.32±7.55)%比(192.63±10.42)%]降低(P均< 0.05),BCL2蛋白水平、SOD [(72.33± 5.87)、(92.08±6.27)、(116.38±7.66)比(52.14±4.95)U/mg]水平、miR-190a-5p表达量升高(P < 0.05),且呈浓度依赖性;与H/R+miR-NC组比较,H/R+miR-190a-5p组细胞凋亡率[(13.51± 1.65)%比(33.07±2.52)%]、cleaved caspase-3、MDA [(13.15±1.67)比(24.08± 3.24) nmol/ mg]、LDH [(29.48±2.55)比(50.68±4.83) U/L)]、ROS [(132.14±9.47)%比(180.11± 12.45)%]水平降低(P均< 0.05),BCL2蛋白水平、SOD [(85.64±6.11)比(51.27± 4.98)U/mg]水平升高(P均< 0.05);转染anti-miR-190a-5p可减低TFA对H/R诱导的H9c2细胞凋亡、氧化应激的作用。结论:TFA可通过上调miR-190a-5p表达而抑制细胞凋亡、氧化应激从而减轻H/R诱导的H9c2细胞损伤。
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Key words
黄芪总黄酮,miR-190a-5p,缺氧/复氧,心肌细胞H9c2,大鼠,细胞凋亡
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