【作者】 陈爽；

【导师】 李孝生；

【作者基本信息】 重庆医科大学 ， 内科学， 2009， 硕士

【摘要】 目的:研究川芎嗪(TMP)对大鼠肝星状细胞(HSC-T6)结缔组织生长因子(CTGF)及Smad2/3表达的影响。方法:体外培养HSC-T6细胞株,以不同浓度的TMP以及0.005mg/l[1]的转化生长因子-β1(TGF-β1)作用于HSC-T6,采用MTT比色法测定HSC-T6增殖;用免疫细胞化学染色法观察CTGF、Smad2/3在HSC-T6的定位及不同浓度TMP对其表达的影响;用Western印迹法检测CTGF、Smad2/3蛋白表达水平。结果:1.一定浓度(100、200、400、600、1000 mg/l) TMP能抑制HSC-T6增殖,且呈剂量依赖性(P<0.05)。2.免疫细胞化学法结果显示CTGF主要在胞浆表达;Smad2/3蛋白则在胞核及胞浆均有表达。随着TMP浓度的升高,CTGF、Smad2/3表达依次递减,与空白对照组比较,具有显著性差异(P<0.05)。3. Western印迹法结果亦显示TMP可以明显抑制CTGF及Smad2/3蛋白的表达,两者被抑制程度呈正相关关系(r=0.967,P<0.01)。结论:TMP可能通过抑制HSC-T6细胞增殖,下调Smad2/3、CTGF的表达,从而发挥其抗肝纤维化的作用。更多还原

【Abstract】 Objective: To study the effects of Tetramethylpyrazine(TMP) on expression of connective tissue growth factor(CTGF) and Smad2/3 in hepatic stellate cell-T6 (HSC-T6).Methods:HSC-T6 was cultured with varying doses of TMP and 0.005mg/l[1] of transforming growth factor-β1(TGF-β1) in vitro.MTT colorimetric assay was used for detecting the proliferation inhibition ratio of HSC-T6.Immunocytochemistry was used to observe the location of CTGF and Smad2/3 ,to test the effect of TMP on the expression of CTGF and Smad2/3 in HSC-T6.Protein expression level of CTGF and Smad2/3 was analyzed with Western blot.Results:1. TMP inhibited the proliferation of HSC-T6 at the concentration of 100、200、400、600 and 1000mg/l in concentration dependent way(P<0.05).2. The test by immunohistochemical method indicated that the expression of CTGF located in the endochylema of HSC-T6 while Smad2/3 expressed both in the nucleus and endochylema of HSC-T6. When the density of TMP increased, the expression of CTGF and Smad2/3 decreased progressively. The comparision between the control group were remarkably discrepant (P<0.05).3. The results of Western blot also revealed that TMP could dose- dependently inhibit the protein expression of CTGF and Smad2/3,the inhibition drgree of which showed a positive correlation.(r=0.967,P<0.01).Conclusion:Inhibiting proliferation of HSC-T6, down-regulating the protein expression of Smad2/3 and CTGF,might be involved in the anti-fibrosis mechanism of TMP.更多还原