摘要
研究川芎嗪对大鼠肝星状细胞株(hepatic stellate cell-T6,HSC-T6)结缔组织生长因子CTGF(connective tissue growth factor)及Ⅰ型胶原表达的影响.培养HSC-T6细胞,不同浓度川芎嗪与转化生长因子-β1(transforming growth factor-β1,TGF-β1)刺激的HSC-T6共同孵育.用MTT(3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)法检测HSC-T6增殖;免疫细胞化学法观察川芎嗪对CTGF表达的影响;Western印迹法检测CTGF蛋白的表达.ELISA(enzyme linked immunosorbent assay)法测定Ⅰ型胶原表达.结果表明,一定浓度(100、200、400、600mg/L)川芎嗪能抑制HSC-T6增殖,且呈剂量依赖性.免疫细胞化学结果显示随着川芎嗪浓度的升高,CTGF表达依次递减,与正常对照组比较,具有显著性差异(t=4.216,P<0.01).川芎嗪还可以明显抑制CTGF蛋白的表达,并可抑制I型胶原合成,二者抑制程度呈正相关关系(r=0.861,P<0.01).川芎嗪可能通过抑制HSC-T6细胞增殖,下调CTGF的表达,阻断Ⅰ型胶原合成,从而发挥其抗肝纤维化的作用.
To study the effects of tetramethylpyrazine（TMP） on expression of connective tissue growth factor （CTGF） and collagen I in hepatic stellate cell-T6 （HSC-T6）. HSC-T6 were cultured with TMP at different concentration after transforming growth factor-β1 （TGF-β1） stimulation. MTT（3-（4,5-Dimethyhhiazol-2-yl）- 2,5-diphenyhetrazolium bromide） colorimetric assay was used for detecting the proliferation inhibition ratio of HSC-T6. Immunocytochemistry was used to test the effect of TMP on the expression of CTGF in HSC-T6. Protein expression level of CTGF was analyzed with Western blot. The expression of collagen I was measured by ELISA （enzyme linked immunosorbent assay） method. Results showed： TMP inhibited the proliferation of HSC-T6 at the concentration of 100, 200, 400 and 600 mg/L in a concentration dependent way pattern. The test by immunohistochemical method indicated that the expression of CTGF decreased progressively when the TMP density increased. The comparison between the control.group were remarkably discrepant （t=4.216, P〈0.01）. TMP also dose-dependently inhibited the protein expression of CTGF and decreased the synthesis of collagen I , the 0.861, P〈 0.01）. Inhibiting proliferation of decreasing the synthesis of collagen I , might inhibition drgree of which showed a positive correlation （r= HSC-T6, down-regulating the protein expression of CTGF and be involved in the anti-fibrosis mechanism of TMP.
出处
《生命科学研究》
CAS
CSCD
2009年第2期146-151,共6页
Life Science Research
基金
重庆市科委自然科学基金资助项目(2006B135428)
关键词
川芎嗪
肝星状细胞
结缔组织生长因子
Ⅰ型胶原
转化生长因子-Β1
tetramethylpyrazine
hepatic stellate cells
connective tissue growth factor
collagen I
transforming growth factor-β1
