阿昔莫司介导自噬途径保护高糖高脂诱导的心肌细胞损伤

Acipimox mediates autophagy pathway to protect myocardial cell damage induced by high glucose and fat

目的 探讨阿昔莫司对高糖高脂诱导的心肌细胞凋亡的影响及其作用机制。方法 将H9C2细胞分为对照组(正常培养)、溶剂组(高脂溶剂+0.1%二甲基亚砜)、高脂高糖组(0.1 mmol·L^(-1)的棕榈酸和50 mmol·L^(-1)的葡萄糖)、低剂量组(高脂高糖诱导+5μg·L^(-1)的阿昔莫司)、中剂量组(高脂高糖诱导+10μg·L^(-1)的阿昔莫司)、高剂量组(高脂高糖诱导+25μg·L^(-1)的阿昔莫司)。用细胞计数试剂盒-8(CCK-8)法检测各组细胞增殖情况;用流式细胞术和TUNEL法检测各组细胞凋亡情况;用蛋白质印迹(Western blot)法检测各组细胞蛋白表达水平;并检测各组细胞氧化应激相关指标水平。结果 对照组、溶剂组、高脂高糖组、低剂量组、中剂量组、高剂量组细胞48 h细胞(光密度值)分别为1.34±0.11、1.29±0.12、0.43±0.05、0.52±0.06、0.79±0.07和0.99±0.10;TUNEL阳性细胞率分别为(3.79±0.38)%、(3.86±0.43)%、(31.27±3.17)%、(25.13±2.10)%、(20.04±2.25)%和(8.34±0.59)%;活性氧水平分别为(10.31±0.79)%、(10.29±0.84)%、(70.12±6.41)%、(61.97±5.12)%、(51.31±4.12)%和(48.97±3.49)%;LC3 II/LC3 I蛋白水平分别为1.10±0.11、1.09±0.12、0.32±0.04、0.45±0.06、0.64±0.07和0.91±0.08。高脂高糖组上述指标分别与对照组、溶剂组比较,差异均有统计学意义(均P<0.05);高脂高糖组上述指标与低剂量组、中剂量组、高剂量组比较,差异均有统计学意义(均P<0.05)。结论 阿昔莫司可通过抑制氧化应激、提高自噬活性减轻高脂高糖诱导的心肌细胞凋亡。

ObjectiveTo investigate the effect of acimolus on cardiomyocyte apoptosis induced by high glucose and fat and its mechanism.Methods H9C2 cells were divided into control group(normal culture),vehicle group (high fat solvent+0.1%dimethyl sulfoxide),HGPA group (0.1 mmol·L^(-1)palmitic acid and 50 mmol·L^(-1)glucose treatment),HGPA+L-ACI group(HGPA group+5μg·L^(-1)acipimox),HGPA+M-ACI group (HGPA group+10μg·L^(-1)acipimox) and HGPA+H-ACI group (HGPA group+25μg·L^(-1)acipimox).Cell proliferation was detected by cell counting kit-8 (CCK-8) method.Flow cytometry and TUNEL were used to detect cell apoptosis in each group.The protein expression levels of each group were detected by Western blot.The levels of oxidative stress related indexes in each group were detected.Results The 48 h cell viability (optical density) in control group,vehicle group,HGPA group,HGPA+L-ACI group,HGPA+M-ACI group and HGPA+H-ACI group were 1.34±0.11,1.29±0.12,0.43±0.05,0.52±0.06,0.79±0.07 and 0.99±0.10,respectively;TUNEL positive cells rate were (3.79±0.38)%,(3.86±0.43)%,(31.27±3.17)%,(25.13±2.10)%,(20.04±2.25)%and (8.34±0.59)%,respectively;reactive oxygen species levels were (10.31±0.79)%,(10.29±0.84)%,(70.12±6.41)%,(61.97±5.12)%,(51.31±4.12)%and (48.97±3.49)%,respectively;the protein levels of LC3Ⅱ/LC3Ⅰwere 1.10±0.11,1.09±0.12,0.32±0.04,0.45±0.06,0.64±0.07 and 0.91±0.08,respectively.Compared with control group and vehicle group,HGPA group showed statistically significant differences in the above indexes (all P0.05);compared with the HGPA group,the above indexes in HGPA+L-ACI group,HGPA+M-ACI group and HGPA+H-ACI group were statistically significant (all P0.05).Conclusion Acipimox can reduce myocardial cell apoptosis induced by high fat and high sugar by inhibiting oxidative stress and improving autophagy activity.