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刘西霞,黄政德,卢芳国,高强,赖娟,芦俊,向琴,刘水平,杨胜辉.抗纤灵Ⅰ方对小鼠血吸虫病肝纤维化的干预及机制探讨[J].湖南中医药大学学报英文版,2016,36(10):11-17.[Click to copy
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| 抗纤灵Ⅰ方对小鼠血吸虫病肝纤维化的干预及机制探讨 |
| 刘西霞,黄政德,卢芳国,高强,赖娟,芦俊,向琴,刘水平,杨胜辉 |
| (中南大学湘雅医学院医学微生物学教研室, 湖南 长沙 410013;长沙医学院医学微生物学与免疫学教研室, 湖南 长沙 410219;湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察抗纤灵Ⅰ方对小鼠血吸虫病肝纤维化的干预效果，探讨抗纤灵Ⅰ方干预血吸虫肝纤维化的可能分子机制，为抗纤灵Ⅰ方的临床应用提供实验依据。方法 采用日本血吸虫尾蚴经皮肤贴片感染昆明小鼠，建立血吸虫病早期肝纤维化动物模型，将建模成功的小鼠用吡喹酮药物连续杀虫治疗后分为抗纤灵Ⅰ方低、中、高剂量组，抗纤灵Ⅰ方的丹参加味方低、中、高剂量组，秋水仙素治疗对照组，蒸馏水灌胃对照组和未做任何处理的模型对照9组。以未感染血吸虫的正常小鼠为肝纤维化阴性对照组。各实验组连续灌胃治疗8周，收集各组小鼠肝脏，HE染色观察肝纤维化病理改变，测量肝组织虫卵肉芽肿平均面积；免疫组化染色方法检测肝组织内胶原蛋白Ⅰ、Ⅲ、MMP-1蛋白酶以及TIMP-1蛋白的表达，比较各组间的蛋白表达差异。结果 HE染色显示，与模型组和正常对照组相比抗纤灵Ⅰ方及其丹参加味方高剂量组可显著减少肝组织虫卵肉芽肿面积（P<0.05）；免疫组化染色结果显示，与模型组比较，抗纤灵Ⅰ方及其丹参加味方中、高剂量组干预后小鼠肝组织内的胶原Ⅰ、Ⅲ蛋白以及TIMP-1蛋白表达显著减少（P<0.05）、MMP-1蛋白酶表达显著增加（P<0.05），但抗纤灵Ⅰ方与丹参加味方之间差异无统计学意义（P>0.05）。结论 抗纤灵Ⅰ方及其丹参加味方对小鼠血吸虫病肝纤维化具有较好的干预效果，但两者在疗效上差异无统计学意义，丹参未能增强抗纤灵Ⅰ方的干预肝纤维化效果；抗纤灵Ⅰ方的干预机制可能是通过抑制肝组织内胶原蛋白Ⅰ、Ⅲ和TIMP-1蛋白的表达，增加MMP-1蛋白酶的表达来发挥作用。 |
| 关键词: 抗纤灵Ⅰ方 血吸虫病 肝纤维化 胶原蛋白Ⅰ 胶原蛋白Ⅲ MMP-1蛋白酶 TIMP蛋白 丹参 |
| DOI：10.3969/j.issn.1674-070X.2016.10.004 |
| Received:May 11, 2016 |
| 基金项目:国家自然基金项目（81373576）；湖南省自然科学基金项目（11JJ3106）；湖南省中医药管理局重点项目（201209）；湖南省卫生厅基金项目（B2014-043）；湖南省高校感染性疾病中医药防治研究科技创新团队开放基金项目（Grxjb-3）；湖南省教育厅基金项目（15C0147）。 |
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| The Intervention Effects and Mechanisms of KangXianLing PrescriptionⅠon Schistosomiasis Liver Fibrosis in Mice |
| LIU Xixia,HUANG Zhengde,LU Fangguo,GAO Qiang,LAI Juan,LU Jun,XIANG Qin,LIU Shuiping,YANG Shenghui |
| (Department of Medical Microbiology, Xiangya School of Medicine, Central South University, Changsha, Hunan 410013, China;Department of Medical Microbiology and Immunology, Changsha Medical University, Changsha, Hunan 410219, China;Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the intervention effects of KangXianLing prescriptionⅠ(KXLPI) on schistosomiasis liver fibrosis in mice,elucidate the possible molecular mechanisms of KXLPⅠ anti hepatic fibrosis,and provide experimental basis for clinical application of KXLPⅠ.Methods Kunming mice were infected by Sj cercariae in order to establish the animal model with hepatic fibrosis.All the model mice with hepatic fibrosis were treated by praziquantel (PZQ) and randomly divided into 9 groups,including KXLPⅠlow dose group (L-KXLPⅠ),medium dose group (M-KXLPⅠ) and high dose group (H-KXLP I),KXLPⅠ's modified prescription based on Salvia miltiorrhiza (KXLPⅠ-Sm) low dose group (L-KXLPⅠ-Sm),medium dose group (M-KXLPⅠ-Sm) and high dose group (H-KXLPⅠ-Sm),Colchicin treatment group (CTG),distilled water group (DWG) and model group (MG),and the mice which were not infected by Sj cercariae are used as normal control group (NG).After intragastric administration for 8 weeks,the livers of the mice were collected and the pathologic changes of hepatic fibrosis were observed by HE.The mean granuloma areas induced by eggs were detected,and pathological changes of hepatic fibrosis were observed.The expression levels of collagen Ⅰ,Ⅲ,matrix-metalloproteinase-1(MMP-1) and tissue inhibitor of metalloproteinase-1(TIMP-1) were detected by immunohistochemical staining,the differences of proteins were compared.Results Compared with MG and NG,the mean granuloma areas in H-KXLPⅠand H-KXLPⅠ-Sm significantly decreased,(P<0.05).Immunohistochemical staining results showed that the expression levels of collagen Ⅰ,Ⅲ and TIMP-1 in M-KXLPⅠ,H-KXLPⅠ,M-KXLPⅠ-Sm and H-KXLPⅠ-Sm were significantly lower than that of MD group (P<0.05),while the expression levels of MMP-1 significantly higher than that of MD (P<0.05),there is no difference between KXLPⅠand KXLPⅠ-Sm groups (P>0.05).Conclusion KXLPⅠand KXLPⅠ-Sm showed good preventive effects on experimental schistosomiasis liver fibrosis mice,but there was no difference on curative effect,and Salvia miltiorrhiza could not enhance the effect of KXLPⅠanti hepatic fibrosis.Meanwhile,KXLPⅠcould inhibit the expressions of collagenⅠ,Ⅲ and TIMP-1 in liver tissues and increase the expression of MMP-1. |
| Key words: KangXianLing Prescription I schistosomiasis hepatic fibrosis collagen Ⅰ collagen Ⅲ matrix-metal loproteinase-1 tissue inhibitor of metalloproteinase-1 Salvia miltiorrhiza |
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