三七皂苷R2对棕榈酸诱导H9c2心肌细胞脂毒性的影响

Effects of notoginsenoside R2 on palmitic acid-induced lipotoxicity in H9c2 cardiomyocytes

目的 研究三七皂苷R2(NGR2)对棕榈酸(PA)诱导H9c2心肌细胞脂毒性的影响。方法 将H9c2心肌细胞分为对照组、模型组和低、高剂量实验组。对照组给予10%牛血清白蛋白(BSA)处理;模型组给予250μmol·L^(-1)PA以建立脂毒性模型;低、高剂量实验组分别给予50、100μmol·L^(-1)NGR2和250μmol·L^(-1)PA处理。用油红O和Bodipy染色法观察细胞脂滴积累情况,用实时荧光定量聚合酶链反应(RT-qPCR)法检测脂质代谢相关的脂肪酸合成酶(FASN)、硬脂酰辅酶A去饱和酶1(SCD1) mRNA相对表达水平,用蛋白质印迹法检测FASN、SCD1蛋白的相对表达水平。结果 对照组、模型组和低、高剂量实验组的油红O染色后脂滴定量光密度值分别为0.12±0.03、0.19±0.02、0.14±0.05和0.11±0.03,Bodipy染色定量相对荧光强度分别为(100.00±15.83)%、(249.65±6.44)%、(160.88±17.34)%和(136.84±4.83)%,FASN mRNA相对表达水平分别为1.00±0.07、1.33±0.13、1.21±0.16和1.05±0.09,SCD1 mRNA相对表达水平分别为1.00±0.04、1.26±0.07、1.24±0.15和1.02±0.09,FASN蛋白相对表达水平分别为1.00±0.00、1.03±0.02、1.01±0.06和0.92±0.08,SCD1蛋白相对表达水平分别为1.00±0.00、1.08±0.08、0.98±0.15和0.87±0.18。模型组的上述指标与对照组比较,高剂量实验组的上述指标与模型组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01);低剂量实验组的油红O和Bodipy染色定量与模型组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01)。结论 NGR2可能通过降低FASN、SCD1 mRNA和蛋白表达,减少脂质积累,减轻PA诱导H9c2心肌细胞的脂毒性。

Objective To study the effect of notoginsenoside R2(NGR2)on palmitic acid(PA)-induced lipotoxicity in H9c2 cardiomyocytes.Methods The H9c2 cells were divided into control group,model group and experimental-L,-H groups.The control group was treated with 10%bovine serum albumin(BSA);the model group was treated with 250μmol·L^(-1) PA to establish the lipotoxicity model;the experimental-L,-H groups were treated with 50,100μmol·L^(-1) NGR2 and 250μmol·L^(-1) PA.The accumulation of cellular lipid droplets was observed by oil red O staining and Bodipy staining;the mRNA expression levels of fatty acid synthase(FASN),stearoyl coenzyme A desaturase 1(SCD1)were detected by real time quantitative polymerase chain reaction;the protein expression levels of FASN and SCD1 were detected by Western blot.Results The absorbance of lipid droplet quantification after oil red O staining in the control group,the model group,the experimental-L,-H groups were 0.12±0.03,0.19±0.02,0.14±0.05 and 0.11±0.03;the relative fluorescence intensity quantified by Bodipy staininggwere(100.00±15.83)%,(249.65±6.44)%,(160.88±17.34)%and(136.84±4.83)%;FASN mRNA expression levels were 1.00±0.07,1.33±0.13,1.21±0.16 and 1.05±0.09;SCD1 mRNA expression levels were 1.00±0.04,1.26±0.07,1.24±0.15 and 1.02±0.09;the expression levels of FASN protein were 1.00±0.00,1.03±0.02,1.01±0.06 and 0.92±0.08;the expression levels of SCD1 protein were 1.00±0.00,1.08±0.08,0.98±0.15 and 0.87±0.18.In the above indicators,there were statistically significant differences between model group and control group,between experimental-H group and model group(P<0.05,P<0.01);there were significant differences in oil red O staining and Bodipy staining quantification of experimental-L group and model group(P<0.05,P<0.01).ConclusionN IGR2 may reduce lipid accumulation and alleviate lipotoxicity in PA-induced H9c2 cardiomyocytes by decreasing the mRNA and protein expression of FASN and SCD1.