中药复方对血吸虫性肝纤维化小鼠肝组织MMP-2及TIMP-2表达的影响
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摘要
目的:探讨中药复方对血吸虫性肝纤维化小鼠肝组织基质金属蛋白酶-2(matrix metalloproteinase-,MMP-2)及基质金属蛋白酶的组织抑制因子-2(tissue Inhibitor of Matrix Metalloproteinase-2,TIMP-2)表达的影响。方法:将108只小鼠按照随机数字表法分为中药复方I组、中药复方J组、中药复方K组、中药复方L组、空白模型组及对照组,每组各18只。采用血吸虫尾蚴经皮感染小鼠腹部的方法将中药复方I组、中药复方J组、中药复方K组、中药复方L组、空白模型组小鼠制备肝纤维化模型。将基于毒、痰、瘀、虚四大证候要素进行组方的中药复方分别灌胃中药复方I组、中药复方J组、中药复方K组、中药复方L组小鼠,空白模型组与对照组灌胃生理盐水。HE染色后,光镜下观察小鼠肝脏病理切片肝纤维化程度; Masson染色后,采用SABC免疫组织化学技术半定量观察各组小鼠肝脏病理切片中肝组织表达MMP-2、TIMP-2的变化。结果:中药复方L组MMP-2表达最高,TIMP-2表达最低,与中药复方K组、中药复方J组、中药复方I组、空白模型组比较,差异均有统计学意义(P <0. 05)。病理结果显示,中药复方L组小鼠的肝脏纤维化程度最低,中药复方K组、中药复方J组、中药复方I组、空白模型组小鼠的肝脏纤维化程度依次加重。结论:血吸虫性肝纤维化小鼠具有毒、痰、瘀、虚的证候要素特点,中药复方能明显改善血吸虫性肝纤维化小鼠肝组织中MMP-2和TIMP-2的含量。
Objective: To explore the influence of TCM compound formula based on the theory of toxic and phlegm and blood stasis and deficiency on the expression of matrix metalloproteinase-2( MMP-2) and tissue Inhibitor of Matrix Metalloproteinase-2( TIMP-2) in liver tissue of mice with schistosomiasis hepatic fibrosis. Methods: A total of 108 mice were randomly divided into the TCM compound formula group I,the TCM compound formula group J,the TCM compound formula group K,the TCM compound formula group L,the blank model group and the control group,with 18 mice in each group. Hepatic fibrosis model was established by transdermal infection of schistosomiasis cercariae in mice abdomen. Based on the four syndromes of toxic and phlegm and blood stasis and deficiency,the mice in the group I,the group J,the group K and the group L were given TCM compound formula by gavage,while the mice in the blank model group and the control group were given normal saline by gavage. After HE staining,the degree of hepatic fibrosis was observed under light microscope. After Masson staining,the expression of MMP-2 and TIMP-2 in liver tissue of mice in each group was semi-quantitatively observed by SABC immunohistochemical technique. Results:The expression of MMP-2 was the highest in group L,while TIMP-2 was the lowest and the ratio of MMP-2 to TIMP-2 was the highest There were significant differences between the group L and the group K,the group J,the group I and the blank model group( P < 0. 05). Pathological results showed that the hepatic fibrosis of mice in the group L was the lowest,and the hepatic fibrosis of mice in the group K,the group J,the group I and the blank model group was aggravated in turn. Conclusion: Mice with schistosomiasis hepatic fibrosis have the characteristics of syndrome elements of toxic and phlegm and blood stasis and deficiency.TCM compound formula can significantly improve the pathological changes of liver tissue,and up-regulate the ratio of MMP-2 to TIMP-2.
Objective: To explore the influence of TCM compound formula based on the theory of toxic and phlegm and blood stasis and deficiency on the expression of matrix metalloproteinase-2( MMP-2) and tissue Inhibitor of Matrix Metalloproteinase-2( TIMP-2) in liver tissue of mice with schistosomiasis hepatic fibrosis. Methods: A total of 108 mice were randomly divided into the TCM compound formula group I,the TCM compound formula group J,the TCM compound formula group K,the TCM compound formula group L,the blank model group and the control group,with 18 mice in each group. Hepatic fibrosis model was established by transdermal infection of schistosomiasis cercariae in mice abdomen. Based on the four syndromes of toxic and phlegm and blood stasis and deficiency,the mice in the group I,the group J,the group K and the group L were given TCM compound formula by gavage,while the mice in the blank model group and the control group were given normal saline by gavage. After HE staining,the degree of hepatic fibrosis was observed under light microscope. After Masson staining,the expression of MMP-2 and TIMP-2 in liver tissue of mice in each group was semi-quantitatively observed by SABC immunohistochemical technique. Results:The expression of MMP-2 was the highest in group L,while TIMP-2 was the lowest and the ratio of MMP-2 to TIMP-2 was the highest There were significant differences between the group L and the group K,the group J,the group I and the blank model group( P < 0. 05). Pathological results showed that the hepatic fibrosis of mice in the group L was the lowest,and the hepatic fibrosis of mice in the group K,the group J,the group I and the blank model group was aggravated in turn. Conclusion: Mice with schistosomiasis hepatic fibrosis have the characteristics of syndrome elements of toxic and phlegm and blood stasis and deficiency.TCM compound formula can significantly improve the pathological changes of liver tissue,and up-regulate the ratio of MMP-2 to TIMP-2.
引文
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[10]谭勤锐,李晖,杨琪,等.基质金属蛋白酶及其特定抑制剂在肝纤维化形成中的作用研究进展[J].山东医药,2017,57(34):109-111.
[11]戈宏焱,张仕华.基质金属蛋白酶与肝纤维化关系的研究进展[J].临床肝胆病杂志,2017,33(3):563-566.
[2]张梦然,成军.肝纤维化研究进展[J].国际消化病杂志,2014,34(6):374-379.
[3]赵双双,邵荣光,何红伟.抗肝纤维化的潜在作用靶点[J].药学学报,2014,49(10):1365-1371.
[4]谢玉梅,聂青和,周永兴,等.肝硬化患者肝组织中TIMP-1,TIMP-2的表达[J].第四军医大学学报,2000,21(7):790-792.
[5]梁志清.细胞外基质的降解代谢与肝纤维化[J].国外医学·生理病理科学与临床分册,1998,18(4):352-354.
[6]IREDALE J P.Tissue inhibitors of metalloproteinases in liver fibrosis[J].International Journal of Biochemistry&Cell Biology,1997,29(1):43-54.
[7]OLASO E,FRIEDMAN S L.Molecular regulation of hepatic fibrogenesis[J].Journal of Hepatology,1998,29(5):836-847.
[8]陶然,黄加权,李小丹,等.热休克蛋白47-短发夹RNA对日本血吸虫肝纤维化小鼠的体内干预研究[J].中华传染病杂志,2014,32(7):391-396.
[9]陈珂,许君望,周琦,等.全反式维甲酸对TGF-β1刺激的肝星状细胞COL1α2、MMP-2、TIMP-1以及信号通路的影响[J].西安交通大学学报(医学版),2017,38(6):857-861.
[10]谭勤锐,李晖,杨琪,等.基质金属蛋白酶及其特定抑制剂在肝纤维化形成中的作用研究进展[J].山东医药,2017,57(34):109-111.
[11]戈宏焱,张仕华.基质金属蛋白酶与肝纤维化关系的研究进展[J].临床肝胆病杂志,2017,33(3):563-566.