木犀草素对TGF-β1诱导HSC-T6增殖凋亡及分泌α-SMA、CollagenⅠ影响  

Effect of luteolin on TGF-β1 induced proliferation and apoptosis of HSCT6 andα-SMA and collagenⅠsecretion

作　　者：韩志强[1] 安达[1] 巴图德力根[1] HAN Zhi-Qiang;AN Da;BA TU-De-Li-Gen(Affiliated Hospital of Inner Mongolia Minzu University&Institute of Clinical Pharmacology of Traditional Mongolian Medicine,Tongliao 028000,Inner Mongolia,China)

机构地区：[1]内蒙古民族大学附属医院蒙药临床药理研究所,内蒙古通辽028000

出　　处：《中国老年学杂志》2024年第16期3929-3932,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(81760908)。

摘　　要：目的观察木犀草素对转化生长因子(TGF)-β1诱导HSC-T6增殖、凋亡及分泌α-平滑肌肌动蛋白(SMA)、胶原纤维(Collagen)Ⅰ的影响。方法取对数生长期细胞,接种于6孔板中,设空白组、TGF-β1低、中、高剂量组,每个浓度设3个复孔,TGF-β1低、中、高剂量组分别用100、200、400 ng TGF-β1处理48 h。细胞传代扩大培养至所需的细胞量后,分为对照组、模型组(TGF-β150 ng/ml)、木犀草素高、中、低剂量组(TGF-β150 ng/ml+10.000、2.000、0.667μmol/L木犀草素)、HSC-T6+TGF-β1组、HSC-T6+TGF-β1+木犀草素组(40μmol/L木犀草素)。酶联免疫吸附试验(ELISA)检测α-SMA、CollagenⅠ、Ⅲ含量,用实时荧光定量聚合酶链反应(RT-PCR)检测细胞α-SMA、CollagenⅠ、Ⅲ、基质金属蛋白酶(MMP)-1、MMP抑制剂(TIMP)-1 mRNA表达。TGF-β1诱导HSC-T6建立肝纤维化模型,给予木犀草素干预,噻唑蓝(MTT)法检测HSC-T6细胞增殖,ELISA检测α-SMA、CollagenⅠ含量,流式细胞技术检测HSC-T6凋亡。结果与空白组比较,TGF-β1低、中、高剂量组Col-lagenⅠ、ⅢmRNA及蛋白、α-SMA、TIMP-1 mRNA表达升高,TGF-β1低、高剂量组α-SMA表达升高,TGF-β1高剂量组MMP-1 mRNA表达升高,差异有统计学意义(均P<0.05)。与对照组比较,模型组α-SMA、CollagenⅠ含量明显升高(P<0.05);与模型组比较,木犀草素中、高剂量组增殖率明显降低,木犀草素高剂量组α-SMA、CollagenⅠ含量明显降低(P<0.05)。与HSC-T6+TGF-β1组比较,HSC-T6+TGF-β1+木犀草素组早期、晚期凋亡细胞明显增加(P<0.05)。结论木犀草素具有抑制HSC-T6细胞分泌胶原蛋白,抑制HSC-T6细胞增殖,促进早期凋亡、晚期凋亡的作用。Objective To observe the effect of luteolin on transforming growth factor(TGF)-β1 induced the proliferation and apoptosis of HSC-T6 and the secretion ofα-smooth muscle actin(SMA)and CollagenⅠ.Methods Logarithmic growth cells were in-oculated in 6-well plates,the blank group and low,medium,high dose of TGF-β1 groups were set,with 3 secondary wells for each concentration.Low,medium and high dose of TGF-β1 groups were treated with 100,200 and 400 ng TGF-β1 for 48 h respectively.After expanding the culture to the required number of cells,cells were divided into control group,model group(TGF-β150 ng/ml),high,medium and low dose of luteolin groups(TGF-β150 ng/ml+10.000,2.000,0.667μmol/L luteolin),HSC-T6+TGF-β1 group and HSC-T6+TGF-β1+luteolin group(40 mmol/L luteolin).Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents ofα-SMA,CollagenⅠandⅢ,real-time fluorescence quantitative polymerase chain reaction(RT-PCR)was used to detect the expressions ofα-SMA,CollagenⅠandⅢ,matrix metalloproteinase(MMP)-1 and MMP inhibitor(TIMP)-1 mRNA.A liver fi-brosis model was established through TGF-β1 induced HSC-T6,and treated with luteolin.The proliferation of HSC-T6 cells was detec-ted by methyl thiazolyl tetrazolium(MTT)assay,the contents ofα-SMA and collagenⅠwere detected by ELISA,and the apoptosis of HSC-T6 was detected by flow cytometry.Results Compared with the blank group,the expressions of CollagenⅠ,ⅢmRNA and pro-tein,α-SMA and TIMP-1 mRNA were increased in low,medium and high dose of TGF-β1 groups,the expression ofα-SMA was in-creased in low and high dose of TGF-β1 groups,and the expression of MMP-1 mRNA was increased in high dose of TGF-β1 group,the difference was statistically significant(all P<0.05).Compared with control group,the contents ofα-SMA and CollagenⅠin model group were significantly increased(P<0.05);compared with model group,the proliferation rate of medium and high dose of TGF-β1 luteolin groups was significantly decreased,and the contents ofα-SMA and Col

关 键 词：转化生长因子(TGF)-β1 肝星状细胞 木犀草素 增殖 凋亡 

分 类 号：R33[医药卫生—人体生理学]