檀香提取物调控miR-199a-3p对大鼠心肌细胞缺氧/复氧损伤的作用

Effect of sandalwood extract on cardiomyocyte hypoxia/reoxygenation injury in rats through regulating miR-199a-3p

目的探讨檀香提取物对心肌细胞缺氧/复氧(H/R)损伤的影响及其分子机制。方法建立心肌细胞H/R模型,并加入不同浓度(0.1,0.2,0.4 mg/ml)的檀香提取物。采用细胞计数试剂盒(CCK-8)检测细胞存活,流式细胞术检测细胞凋亡,商品试剂盒检测乳酸盐脱氢酶(LDH)和肌酸激酶(CK)活性,实时荧光定量PCR(RT-qPCR)检测miR-199a-3p的表达。将心肌细胞H9c2分为H/R+miR-NC组、H/R+miR-199a-3p组、H/R+anti-miR-NC组、H/R+anti-miR-199a-3p、H/R+0.4 mg/ml檀香+miR-NC组、H/R+0.4 mg/ml檀香+miR-199a-3p组、H/R+0.4 mg/ml檀香+anti-miR-NC组、H/R+0.4 mg/ml檀香+anti-miR-199a-3p组。CCK-8法、流式细胞术、商品试剂盒测量细胞存活、凋亡和损伤情况。结果与H/R组比较,H/R+檀香组细胞存活率、miR-199a-3p的表达升高(P<0.05),凋亡率、LDH和CK活性降低(P<0.05)。与H/R+miR-NC组比较,H/R+miR-199a-3p组细胞存活率升高(P<0.05),凋亡率、LDH和CK活性降低(P<0.05)。与H/R+anti-miR-NC组比较,H/R+anti-miR-199a-3p组细胞存活率降低(P<0.05),凋亡率、LDH和CK活性升高(P<0.05)。与H/R+0.4 mg/ml檀香+anti-miR-NC组比较,H/R+0.4 mg/ml檀香+anti-miR-199a-3p组细胞存活率降低(P<0.05),凋亡率、LDH和CK活性升高(P<0.05)。与H/R+0.4 mg/ml檀香+miR-NC组比较,H/R+0.4 mg/ml檀香+miR-199a-3p组细胞存活率升高(P<0.05),凋亡率、LDH和CK活性降低(P<0.05)。结论檀香提取物上调miR-199a-3p对大鼠心肌细胞缺氧/复氧损伤具有保护作用。

Objective To investigate the effect of sandalwood extract on hypoxia/reoxygenation(H/R)injury of rat cardiomyocytes and its molecular mechanism.Methods Cardiomyocyte H/R model was constructed,and then the model cells were treated with different concentrations of sandalwood extract(0.1,0.2,0.4 mg/ml).Cell survival was detected by CCK-8 method.Apoptosis was measured by flow cytometry.LDH and CK activities were tested by commercial kits.And the expression of miR-199a-3p was detected by RT-qPCR.H9c2 cells were divided into H/R+miR-NC group,H/R+miR-199a-3p group,H/R+anti-miR-NC group,H/R+anti-miR-199a-3p,H/R+0.4 mg/ml sandalwood+miR-NC group,H/R+0.4 mg/ml sandalwood+miR-199a-3p group,H/R+0.4 mg/ml sandalwood+anti-miR-NC group,H/R+0.4 mg/ml sandalwood+anti-miR-199a-3p group.Cell survival,apoptosis and LDH and CK acti-vities were measured by CCK-8 method,flow cytometry and commercial kit,respectively.Results Compared with H/R group,the cell survival rate and miR-199a-3p expression in H/R+sandalwood group were increased(P<0.05),while the apoptosis rate,and LDH and CK activities were decreased(P<0.05).Compared with H/R+miR-NC group,the cell survival rate in H/R+miR-199a-3p group was increased(P<0.05),while the apoptosis rate,and LDH and CK activities were decreased(P<0.05).Compared with H/R+anti-miR-NC group,the cell survival rate in H/R+anti-miR-199a-3p group was reduced(P<0.05),while the apoptosis rate,LDH and CK activities were increased(P<0.05).Compared with H/R+0.4 mg/ml sandalwood+anti-miR-NC group,the cell survival rate in H/R+0.4 mg/ml sandalwood+anti-miR-199a-3p group was reduced(P<0.05),while the apoptosis rate and the activities of LDH and CK were increased(P<0.05).Compared with H/R+0.4 mg/ml sandalwood+miR-NC group,the cell survival rate in H/R+0.4 mg/ml sandalwood+miR-199a-3p group was increased(P<0.05),while the apoptosis rate,and LDH and CK activities were decreased(P<0.05).Conclusion Sandalwood extract has protective effect against hypoxia/reoxygenation injury of rat cardiomyocytes by up-regulating miR-199a-3p.