当归红芪超滤物对放射性H9C2细胞损伤TNF-α/CTRP9信号通路相关因子表达的影响

Effects of Ultra-filtration Extracts from Angelicae Sinensis Radix and Hedysari Radix for TNF-α/CTRP9 Signaling Pathway Related Factors in Damaged H9C2 Cell by Radiation

目的观察当归红芪超滤物对放射性H9C2细胞损伤的影响,探讨其对心肌损伤的保护作用机制。方法采用大鼠灌胃方法制备药物血清。将H9C2细胞随机分为空白组、阴性对照组、模型组、盐酸贝那普利组、当归红芪超滤物组和盐酸贝那普利+当归红芪超滤物组。药物血清干预及X线照射H9C2细胞后,ELISA检测肌钙蛋白T(cTnT)、肌钙蛋白Ⅰ(c TnⅠ)及C-反应蛋白(CRP)含量,Western blot和RT-PCR检测肿瘤坏死因子-α(TNF-α)和肿瘤坏死因子相关蛋白9(CTRP9)的表达,免疫荧光检测转化生长因子-β(TGF-β)和α-平滑肌肌动蛋白(α-SMA)的表达。结果与空白组比较,模型组H9C2细胞c Tn T、c TnⅠ、CRP含量明显增加,TNF-α、CTRP9蛋白和基因表达明显升高,α-SMA和TGF-β蛋白表达明显升高;与模型组比较,盐酸贝那普利组、当归红芪超滤物组和盐酸贝那普利+当归红芪超滤物组cTnT、c TnⅠ、CRP含量明显减少,TNF-α、CTRP9蛋白和基因表达明显降低,α-SMA和TGF-β蛋白表达明显降低。结论当归红芪超滤物可通过下调TNF-α/CTRP9信号通路相关因子的表达,达到保护放射性H9C2细胞损伤的目的。

Objective To observe the effects of ultra-filtration extracts from Angelicae Sinensis Radix and Hedysari Radix in damaged H9C2 cell by radiation;To explore its protective mechanism against myocardial injury.Methods The drug serum was made by rat gavage method.H9C2 cells were randomly divided into blank group,negative control group,model group,Benazepril hydrochloride group,ultra-filtration extracts from Angelicae Sinensis Radix and Hedysari Radix group and Benazepril hydrochloride+ultra-filtration extracts from Angelicae Sinensis Radix and Hedysari Radix group.After drug serum intervention and X ray radiation H9C2 cells,the content of cTnT,c TnⅠand CRP were detected by ELISA;Western blot and RT-PCR were used to detect TNF-αand CTRP9 expressions;immunofluorescence staining was used to detect the protein expressions of TGF-βandα-SMA.Results Compared with the blank group,the contents of cTnT,cTnⅠ,CRP in H9C2 cells of model group significantly increased,and the protein and m RNA expressions of TNF-α,CTRP9 significantly increased,protein expressions ofα-SMA and TGF-βsignificantly increased.Compared with the model group,the contents of cTnT,cTnⅠ,CRP in the Benazepril hydrochloride group,ultra-filtration extracts from Angelicae Sinensis Radix and Hedysari Radix group and the Benazepril hydrochloride+ultra-filtration extracts from Angelicae Sinensis Radix and Hedysari Radix group were significantly reduced,protein and mRNA expressions of TNF-α,CTRP9 were significantly reduced,protein expressions ofα-SMA and TGF-βwere significantly reduced.Conclusion Ultrafiltration extracts from Angelicae Sinensis Radix and Hedysari Radix can down-regulate the expressions of TNF-α/CTRP9 signaling pathway related factors to protect damaged H9C2 cells by radiation.