姜黄素上调PPARγ抑制肝星状细胞活化标志表达的研究  

Curcumin inhibits the activation marker of hepatic stellate cells by up-regulating the peroxisome proliferator-activated receptorγ.

作　　者：成扬[1] 平键[1] 徐列明[1] 刘成[1] 谭英姿[1] 陈高峰[1] 

机构地区：[1]上海中医药大学附属曙光医院肝病研究所,上海201203

出　　处：《中国实用内科杂志：临床前沿版》2006年第12期1937-1940,共4页

基　　金：国家自然科学基金(30300458)

摘　　要：目的研究姜黄素对肝星状细胞（HSC）增殖活化的影响与过氧化物酶体增殖因子活化受体γ（PPARγ）之间的关系。方法2005年4月至2006年8月，上海中医药大学曙光医院肝病研究所采用肝脏原位灌流酶消化、Nycodenz密度梯度离心法分离培养大鼠HSC，使用药物处理细胞，MTT法检测药物对细胞增殖的影响。收集裂解细胞抽提细胞总RNA，半定量RT—PCR检测基因表达水平。抽提细胞总蛋白，10％聚丙烯酰胺凝胶电泳分离蛋白，Western blot检测蛋白表达水平。结果姜黄素在10～50μmol／L范围内呈剂量依赖性抑制HSC的增殖。姜黄素对HSC增殖的抑制作用可以被PPAR~t特异性拮抗剂GW9662所阻断。在原代培养第1、4、7天和传代培养的HSC中，PPARγ基因表达水平随着HSC活化程度的增加而不断下降；姜黄素显著上调PPARγ表达水平，这种作用能够被GW9662阻断。姜黄素能够在基因和翻译水平显著抑制α平滑肌肌动蛋白的表达，GW9662能够阻断姜黄素的抑制作用。结论姜黄素通过上调PPARγ抑制大鼠HSC的增殖与活化。Objective To study the relation between the effect of Cureumin on the proliferation and activation of hepatic stellate cells （HSC） and the expression of the peroxisome proliferator - activated receptorγ （PPARγ）. Methods The rat HSC was isolated from SD rats through in situ perfusion of liver with Pronase E and density - gradient centrifugation with Nycodenz. The subeultured cells were treated with COlTesponding compound. The inhibition effect on HSC proliferation was determined by MTr eolorimetry. The total RNA was extracted by TRizol reagent, and the gene expression level of PPARα and smooth muscle actin （ α - SMA） were determined by semi - quantitative RT - PCR. The total cellular proteins were extracted and separated by sodium dodecyl sulfate - polyacrylamide gel electrophoresis, and the protein level of α - SMA was determined by Western blotting. Results The MTT analysis results showed that Curcumin inhibited HSC proliferation between 10 and 50 μmoL/L in a dose - dependent manner. In the Day 1, 4, 7 of primary HSC and passaged HSC, the expression of PPARγ mRNA decreased with HSC activation extent ; Curcumin could up - regulate the expression level of PPARγ. Cureumin suppressed the expression of α - SMA not only at the gene level but also at the translation level. But all these effects of Curcumin on HSC could be blocked by the PPAR~ specific antagonist GW9662. Conclusion The effect of Curcumin on HSC cell proliferation and activation is through the up - regulation of PPARγ signal.

关 键 词：肝星状细胞 姜黄素 过氧化物酶体增殖因子活化受体 Α平滑肌肌动蛋白 

分 类 号：R5[医药卫生—内科学]