miRNA-21和Smad7在砷暴露肝纤维化大鼠肝组织中的表达及药物干预  

Expression of mi RNA-21 and Smad7 in arsenicinduced hepatic fibrosis in rats: Possible mechanism of traditional Chinese herbs for preventing hepatic fibrosis

作　　者：詹洁[1] 吴君[2] 赵雪珂[3] 

机构地区：[1]贵阳医学院附属白云医院健康管理中心,贵州省贵阳市550058 [2]贵阳医学院附属白云医院肝炎实验室,贵州省贵阳市550058 [3]贵阳医学院附属医院感染科,贵州省贵阳市550004

出　　处：《世界华人消化杂志》2014年第34期5249-5257,共9页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30471592~~

摘　　要：目的:观察砷暴露肝纤维化大鼠肝组织中微小RNA(micro RNA,mi RNA)-21和Smad7的表达情况,探讨丹芍化纤胶囊预防砷暴露所致肝纤维化的可能机制.方法:将75只♂SD大鼠随机分为5组,即空白对照组(A组)、砷暴露模型组(B组)、扶正化瘀胶囊预防组(C组)、丹芍化纤胶囊低剂量预防组(D组)、丹芍化纤胶囊高剂量预防组(E组),每组15只.各组大鼠自由饮水,其中A组饮用自来水,B-E组饮用含亚砷酸钠(Na As O2)100 mg/L水.同时,C组大鼠予扶正化瘀胶囊(0.8 g/kg),D组予丹芍化纤胶囊(0.5g/k g),E组予丹芍化纤胶囊(1.0 g/k g)分别灌胃,2次/d,5 d/wk,共16 wk.全自动生化分析仪测定大鼠血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)及天冬氨酸氨基转移酶(aspartate aminotransferase,AST)活性,ELISA法检测肝匀浆透明质酸(hyaluronic acid,HA)、转化生长因子-β1(transforming growth factor-β1,TGF-β1)的水平,HE及Masson染色法观测肝组织病理改变,RT-PCR法检测肝组织mi RNA-21与Smad7 m RNA的表达,免疫组织化学法及Western blot检测肝组织Smad7蛋白的表达.结果:与A组比较,B-E组大鼠血清ALT、AST,肝匀浆HA、TGF-β1显著升高,但C、D、E组显著低于B组(F=129.159、291.878、5316.239、1269.287,P<0.01).B组大鼠肝脏可见明显病理改变甚至肝纤维化形成,而C、D、E组肝脏病变及肝纤维化程度显著轻于B组(F=62.832、68.112,P<0.05或P<0.01).与A组比较,B-E组大鼠肝组织mi RNA-21显著升高,但C、D、E组显著低于B组(F=552.231,P<0.01).与A组比较,B-E组大鼠肝组织Smad7m R N A与蛋白的表达均显著降低,但C、D、E组显著高于B组(F=129.512、68.249、43.435,P<0.05或P<0.01).结论:饮水砷暴露16 wk可导致大鼠肝功能受损甚至引起肝纤维化,其机制可能与上调mi RNA-21、下调Smad7的表达有关,丹芍化纤胶囊对砷暴露致大鼠肝纤维化有潜在的预防作用,可能通过下调mi RNA-21、上调Smad7而实现.AIM： To investigate the expression of micro RNA（mi RNA）-21 and Smad7 in arsenic-induced hepatic fibrosis in rats and to explore the possible mechanism of Danshao Huaxian capsule for preventing arsenic-induced hepatic fibrosis. METHODS： Seventy-five male SD rats were randomly divided into a control group（A）, an arsenic exposure group（B）, a Fuzheng Huayu capsule group（C）, low-（D） and high-dose（E） Danshao Haxian capsule groups（n = 15 for each）. Rats in group A were given tap water, while rats in groups B-E were given 100 mg/L Na As O2 solution. Moreover, rats in groups C-E were respectively administered by gastrogavage with Fuzheng Huayu capsule（0.8 g/kg） or Danshao Haxian capsule（0.5, 1.0 g/kg）, twice per day, 5 d/wk for 16 wk. By the end of the experiment, alanine transaminase（ALT） and aspartate transaminase（AST） in serum were measured with an automatic biochemical analyzer, and levels of hyaluronic acid（HA） and transforming growth factor（TGF）-β1 in liver homogenates were determined by ELISA. Histological changes in the liver were observed after HE and Masson’s trichrome staining. The expression of mi RNA-21 in liver tissue was determined by RT-PCR. The m RNA and protein expression of Smad7 in liver tissue was determined by RT-PCR, immunohistochemistry, and Western blot analysis. RESULTS： Compared with group A, serum activities of ALT and AST, levels of HA and TGF-β1 in liver homogenates were significantly increased in groups B-E, but these indices were significantly lower in groups C-E compared with group B（F = 129.159, 291.878, 5316.239, 1269.287, P 〈 0.01）. Obvious proliferation and widened collagen fibers were seen in liver tissues of group B rats, while the degree of hepatic fibrosis was significantly milder in groups C-E than in group B（F = 62.832, 68.112, P 〈 0.05 or P 〈 0.01）. Compared with group A, the expression of mi RNA-21 was significantly higher in groups B-E, but it was significantly lower in groups C-E than in group B（F =

关 键 词：丹芍化纤胶囊 砷 Micro RNA-21 SMAD7 肝纤维化 

分 类 号：R575.2[医药卫生—消化系统]