摘要
目的:研究截短型TGF-βⅡ型受体蛋白对肝纤维化大鼠的治疗作用。方法:取含有重组截短型TGF-βⅡ型受体载体的大肠杆菌BL21,通过培养繁殖,诱导,纯化得到重组截短型TGF-βⅡ型受体蛋白。使用CCl4诱导的方法来制备肝纤维化大鼠模型,观察截短型TGF-βⅡ型受体蛋白对大鼠肝纤维化的影响。SDS-PAGE检测纯化的截短型TGF-βⅡ型受体蛋白。生化检测血清谷丙转氨酶(glutamic pyruvic transaminase,ALT)及谷草转氨酶(glutamic oxalacetic transaminase,AST)含量。用实时荧光定量PCR(real-time PCR)方法和免疫荧光方法来检测各组大鼠肝组织中的α平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA),1型胶原(collagen1,Col1)和4型胶原(collagen4,Col4)的表达情况。用HE,MASSON,PAS这三种病理学染色的方法来观察各组大鼠肝纤维化的变化。结果:SDS-PAGE检测结果显示,纯化后获得高纯度截断型TGF-βⅡ型受体蛋白。截短型TGF-βⅡ型受体蛋白治疗组可以减少血清中谷丙转氨酶和谷草转氨酶的含量。截短型TGF-βⅡ型受体蛋白治疗组可以降低肝组织中α-SMA、FN、Col1和Col4的mRNA和蛋白质的表达。三种病理学染色的方法显示,与肝纤维化大鼠模型组相比,治疗组的纤维化程度明显减轻。结论:重组截短型TGF-βⅡ型受体蛋白对大鼠肝纤维化的程度有抑制作用,为治疗肝纤维化及其他纤维化疾病提供了一个潜在的治疗手段。
Objective： Examine the anti-fibrotic effect of a novel truncated transforming growth factor-β typeⅡ receptor（ t TGF-βRⅡ） in rat hepatic fibrosis model. Methods： The recombinant truncated type TGF-betaⅡ receptor proteins was obtained from the Escherichia coli BL21 strain containing recombinant truncated TGF-betaⅡ receptor vector,by cultivating,inducing,and purification. The rat model of CCL4-induced hepatic fibrosis was established to assess the effect of the t TGF-βR Ⅱ protein on the treatment of fibrosis. SDS-PAGE detect the purification of truncated TGF-beta Ⅱ receptor proteins. The mRNA expression levels of α-SMA,Col1 and Col4 were detected by real-time PCR analysis in hepatic tissue. The content of serum ALT and AST were checked with automated biochemistry analyzer. The expression of α-SMA,Col1 and Col4 protein in hepatic tissue was detected by immunofluorescence. Liver fibrosis of the rats was evaluated by three histological methods： HE staining,Masson staining and PAS staining. Results： SDS-PAGE detection results show that the purified recombinant truncated TGF-beta Ⅱ receptor proteins was obtained. Histological examination revealed that the t TGF-βR Ⅱprotein prevented progression of hepatic fibrosis. The t TGF-βR Ⅱ protein reduced activities of serum ALT and AST. The t TGF-βRⅡ protein reduced both mRNA and protein expression of α-SMA,Col1 and Col4 in the liver tissue. The results of three histopathological staining shows that the treatment group significantly reduce the degree of liver fibrosis. Conclusion： The t TGF-βRⅡ protein prevented progression of hepatic fibrosis in rat and provided a potential treatment for liver fibrosis and other fibrosis diseases.
出处
《中国生物工程杂志》
CAS
CSCD
北大核心
2014年第12期16-22,共7页
China Biotechnology
基金
国家自然科学基金青年科学基金(81200305)
2011年度黑龙江省教育厅重点研究项目计划(12511z028)资助项目
关键词
肝纤维化
转化生长因子Β
受体
Liver fibrosis Transforming growth factor-β Receptor
