摘要
目的阐明RhoA/Rho激酶在糖尿病大鼠肝纤维化中的作用。方法通过高脂饮食和小剂量链脲佐菌素(STZ,30 mg/kg)腹腔注射建立2型糖尿病大鼠模型。实验分为对照组、糖尿病组和法舒地尔干预组(法舒地尔10 mg/(kg·d),分两次腹腔注射)24周末。测定血糖、血脂、谷草转氨酶和谷丙转氨酶;Masson染色和羟脯氨酸测定评估肝胶原沉积;RT-PCR测定转化生长因子β1(TGF-β1)和结缔组织生长因子(CTGF)mRNA表达;免疫组化测定TGF-β1蛋白表达;Western blot测定肝组织肌球蛋白磷酸酯酶靶点亚单位1磷酸化(p-MYPT1)水平。结果与对照组比较,糖尿病组大鼠血糖、血脂和转氨酶显著增高,肝组织大量胶原沉积,p-MYPT1水平、TGF-β1与CTGF mRNA表达显著上调(P<0.01)。与糖尿病组比较,法舒地尔干预组大鼠转氨酶降低,肝胶原沉积明显减轻,p-MYPT1水平、TGF-β1与CTGF mRNA表达显著下降(P<0.01)。结论高血糖激活了肝组织RhoA/Rho激酶,通过调控TGF-β1/CTGF表达,在糖尿病肝纤维化中起着重要作用。
Objective To determine whether RhoA/Rho-kinase is involved in the pathogenesis of hepatic fibrosis in rats with type 2 diabetes. Methods An animal model of type 2 diabetes was developed by high fat diet combined with intraperitoneal injection of low-dose streptozotocin （ STZ, 30 mg/kg, i. p. ）. At week 24, fasting blood glu cose, triglyeerides, cholesterol, aspartate aminotransferase and alanine aminotransferase were measured. The depo sition of collagen in liver was evaluated by Masson staining and the hydroxyproline determination. The mRNA ex pressions of transforming growth factor （TGF-β1） and eonneetive tissue growth factor （CTGF） in liver tissue were assessed by RT-PCR. Immunohistoehemistry staining for TGF-131 was performed. The phosphorylation of myosin phosphatase target subunit 1 （p-MYPT1） was measured by Western blot analysis. Results Compared with control rats, blood glucose, blood fat, aminotransferase and the deposition of collagen in the liver in the untreated diabetic rats were significantly increased （P 〈0. 01 ） ; p-MYPT1 and TGF-β1 protein, the mRNA expression of TGF-β1 and CTGF was significantly enhanced （ P 〈 O. O1 ）. Compared with untreated diabetic rats, treatment with fasudil signi-fieantly decreased aminotransferase, deposition of collagen, p-MYPT1, mRNA expression of TGF-IM and CTGF （P 〈0. 01 ）. Conclusions Hyperglycemia can activate the RhoA/Rho-kinase which maybe regulates TGF-β1/CTGF expression in liver tissues. RhoA/Rho-kinase plays an importent role in the development of diabetic liver fibrosis.
出处
《基础医学与临床》
CSCD
北大核心
2014年第3期332-338,共7页
Basic and Clinical Medicine
基金
河北省卫生厅重大课题(20090007)
