清肝活血方及其拆方对酒精性肝病大鼠CD14、TLR_4表达的影响  

Effect of Qinggan Huoxue recipe and its separated recipes on the expression of CD14 and TLR_4 in rats with alcoholic liver disease

作　　者：吴涛[1,2] 柳涛[1] 郑培永[1] 邢练军[1] 季光[1] 

机构地区：[1]上海中医药大学脾胃病研究所上海中医药大学附属龙华医院消化内科,上海市200032 [2]上海中医药大学中医方证和系统生物学研究中心,上海市201203

出　　处：《世界华人消化杂志》2008年第30期3372-3380,共9页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30572380;教育部新世纪优秀人才支持计划资助项目;No.NCET07-0563~~

摘　　要：目的:探讨清肝活血方及其拆方对酒精性肝病(alcoholic liver disease,ALD)大鼠肝组织CD14、TLR4表达的影响.方法:♂Wistar大鼠100只随机分成空白组、CCl4组各10只,余80只为造模组,采用复合因素复制ALD模型6wk.CCl4组予微量CCl4腹腔注射,每周2次.造模4wk后将模型组随机分成4组,清肝活血方及其拆方组各15只,余为模型组.予等效剂量清肝活血方及拆方ig2wk.检测血清ALT,AST水平;留取肝脏标本进行HE染色.RT-PCR检测肝组织CD14 mRNA、TLR4 mRNA表达,免疫组织化学染色检测肝组织CD14表达,Western blot检测肝组织TLR4蛋白表达.结果:与模型组比较,清肝活血方及其拆方均可明显改善ALD大鼠肝脏脂肪变及肝脏炎症程度(0.67±0.50,2.15±1.28,1.38±1.06vs4.56±0.73,均P<0.01).清肝活血方能显著降低模型大鼠血清ALT水平(725.65±111.02vs884.68±177.54,P<0.05),清肝方和活血方无明显作用;清肝方、活血方、清肝活血方均可明显降低ALD大鼠血清AST水平(2383.81±888.18,2158.93±922.85,2001.90±519.27vs3210.98±640.63,P<0.01或0.05),组间比较无显著差异.清肝方及清肝活血方能显著降低模型大鼠CD14 mRNA表达(1.46±0.52,1.10±0.40vs2.67±0.66,均P<0.01),活血方作用不明显,清肝活血方优于活血方.清肝方、活血方、清肝活血方均能显著降低模型大鼠肝组织TLR4 mRNA表达(1.91±0.03,2.11±0.03,1.53±0.01vs2.37±0.03,均P<0.01),组间比较无显著差异.清肝活血方显著降低模型大鼠肝组织CD14表达(13392.28±9287.54vs32288.89±15631.03,P<0.01).清肝方、活血方、清肝活血方均能显著降低模型大鼠肝组织TLR4表达(1.06±0.10,1.19±0.05,0.98±0.12vs1.40±0.11,均P<0.01).结论:清肝活血方及其拆方发挥对ALD的防治作用,其作用机制可能与降低CD14、TLR4基因和蛋白的表达有关.AIM： To study the effect of Qingganhuoxue Recipe （QGHXR） （liver-clearing bloodactivating） and its separated recipes on the expression of CD14 and TLR4 in rats with ALD METHODS： One hundred male Wistar rats were randomly divided into three groups, that is, experimental group （n = 80） which were established with compound factors, the control group （n = 10） treated with normal saline, and CCl4 group （n = 10） which were intraperitoneally and continuously injected with CCl4 twice a week. Four weeks later, the experimental group was randomly divided into 4 groups： QGHXR group （n = 15）, Qinggan （liver-clearing） recipe group （QGR, n = 15）, Huoxue （blood -activating） recipe group （HXR, n = 15） and model group （n = 50）. All rats were given suitable drugs for two weeks. The serum levels of ALT and AST were detected. Pathological changes in liver tissues were observed using HE staining. The gene expression of CD14 and TLR4 in the liver were measured using RT-PCR, the protein expression of CD14 using immunohistochemical staining, and the expression of TLR4 using Western blot. RESULTS： Compared with the model group, QGHXR and its separated recipes improved the degree of liver steatosis and inflammation signif- icantly （0.67 ± 0.50, 2.15 ± 1.28, 1.38 ± 1.06 vs 4.56 ± 0.73, all P 〈 0.01）. QGHXR degraded blood level of ALT markedly （725.65 ± 111.02 vs 884.68 ± 177.54 P 〈 0.05）, but QGR and HXR showed no remarkable effect. QGR, HXR and QGHXR reduced the serum level of AST significantly （2383.81 ± 888.18, 2158.93 ± 922.85, 2001.90 ± 519.27 vs 3210.98 ± 640.63, P 〈 0.01 or 0.05）, and there was not significant difference among these groups. QGR and QGHXR down-regulated the expression of CD14 mRNA significantly （1.46 ± 0.52, 1.10 ± 0.40 vs 2.67 ± 0.66, both P 〈 0.01）, but HXR showed no obvious effect. QGR, HXR and QGHXR decreased the expression of TLR4 mRNA significantly （1.91 ± 0.03, 2.11 ± 0.03, 1.53 ± 0.01 vs 2.37 ± 0.03, all P 〈 0.01�

关 键 词：清肝活血方 清肝方 活血方 酒精性肝病 CD14 TLR4 

分 类 号：R285.5[医药卫生—中药学]