蓝莓对免疫性肝纤维化大鼠肝组织TLR4,TLR9表达的影响

目的:探讨蓝莓抗氧化作用对猪血清免疫性肝纤维化大鼠肝组织Toll样受体4(toll-like receptor 4,TLR4)和Toll样受体9(TLR9)蛋白表达的影响.方法:60只♂Wistar大鼠随机分为正常对照组(A组)、模型组(B组)、蓝莓低、中、高剂量防治组(C、D、E组)、复方鳖甲软肝片防治组(F组).除A组外,其余各组均腹腔注射猪血清制备大鼠肝纤维化模型.C、D、E、F组在造模同时分别给予蓝莓原浆或复方鳖甲软肝片灌胃,1次/d,共12wk.第12周末处死大鼠,计算大鼠肝脏指数,行肝脏病理组织学检查,测定各组大鼠血清ALT和AST水平、肝组织匀浆超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)及羟脯氨酸(hydroxyproline,Hyp)含量,免疫印迹法(western blot,WB)和免疫组织化学法检测TLR4和TLR9蛋白质在肝脏组织中的表达.结果:各组大鼠血清ALT、AST水平无明显差异(F=4.274,0.917,P>0.05);与B组比较,D,E组肝组织匀浆SOD活性、GSH含量明显升高(F=6.662,61.472,P<0.05);肝脏指数、MDA及Hyp含量明显降低(F=3.972,42.755和41.265,P<0.05);肝纤维化程度明显减轻(F=63.339,P<0.05),胶原表达减少;E组大鼠肝组织TLR4和TLR9蛋白质表达明显减少(F=6.932和7.534,P<0.05).结论:TLR4和TLR9蛋白质表达增加可能与免疫性肝纤维化的发生发展有关.蓝莓对大鼠免疫性肝纤维化有一定的预防作用.

Intragastric administration of blueberry juice decreases the expression of Toll-like receptors 4 and 9 in rats with immune hepatic fibrosis

AIM: To investigate the effect of blueberry on the expression of Toll-like receptor 4 (TLR4) and toll-like receptor 9 (TLR9) in rats with immune hepatic fibrosis. METHODS: Sixty healthy Wistar rats were randomly divided into six groups: normal control group (group A), hepatic fibrosis model group (group B), low-, medium-, and high-concentration blueberry groups (groups C, D and E), and Fufang Biejia Ruanganpian group (group F). Hepatic fibrosis was induced in rats of groups B-F by intraperitoneally injecting porcine serum once daily for 12 wk. Simultaneously, rats of groups C-F were intragastrically given blueberry juice or Fufang Biejia Ruanganpian for 12 wk. All rats were sacrificed at the end of the experiment. Liver index was calculated. Pathological changes in the hepatic tissue were evaluated by hematoxylin-eosin (HE) staining. Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured. The activity of superoxide dismutase (SOD), the contents of malondialdehyde (MDA), hydroxy-proline (Hyp) and reduced glutathione (GSH) in liver homogenates were determined. The expression of TLR4 and TLR9 in the liver was detected by Western blot. RESULTS: There were no significant differences in serum levels of ALT and AST among various groups (F = 4.274, 0.917, both P > 0.05). Compared to group B, the activities of SOD and contents of GSH in liver homogenates were significantly higher (F = 6.662, 61.472, both P < 0.05); liver index and the contents of Hyp and MDA in liver homogenates were significantly lower (F = 3.972, 42.755, 41.265, all P < 0.05); and the degree of hepatic fibrosis was significantly reduced (F = 63.339, P < 0.05) in groups D and E. The expression of hepatic TLR4 and TLR9 in group E was significantly lower than that in group B (F = 6.932, 7.534, both P < 0.05). CONCLUSION: Higher expression of TLR4 and TLR9 protein may be associated with the occurrence and development of immune hepatic fibrosis. Blueberry has a preventive effect on immune hepatic fibrosis in rats.