蒙古山萝卜酸对实验性肝纤维化大鼠MMP-1、TIMP-1、TGF-β_1和PDGF表达的影响
|
摘要
目的探讨蒙古山萝卜酸(MSA)对肝纤维化大鼠血清MMP-1、TIMP-1与肝脏中TGF-β_1、PDGF表达水平的影响。方法将大鼠分为对照组、模型组、阳性对照组(秋水仙碱1.5mg·kg-1)和实验药物高剂量组、低剂量组(MSA 240和30mg·kg-1),每组10只,除对照组外,皮下注射含1mL·kg-1250mL·L~(-1)四氯化碳的花生油,每隔4d注射1次,共计30次。第4次注射后,阳性对照组和实验药物高、低剂量组开始ig给药,每日1次。实验结束后,麻醉,采血,分离血清,ELISA法测定血清MMP-1、TIMP-1,免疫组化法测定肝脏TGF-β_1、PDGF。结果对照组MMP-1、TIMP-1低活性表达,模型组MMP-1表达水平明显降低(与对照组比较P<0.05),TIMP-1表达水平显著升高(与对照组比较P<0.01);阳性药组MMP-1无明显改变(与模型组比较P>0.05),TIMP-1表达水平显著降低(与模型组比较P<0.05);实验药物高剂量组MMP-1表达水平明显上升(与模型组比较P<0.05),TIMP-1表达水平显著降低(与模型组比较P<0.05),MMP-1/TIMP-1比值提高;实验药物低剂量组对MMP-1、TIMP-1无明显变化(与模型组比较均P>0.05)。对照组TGF-β_1、PDGF表达低,模型组TGF-β_1、PDGF表达水平明显升高(与对照组比较均P<0.01);阳性药组TGF-β_1、PDGF表达水平显著降低(与模型组比较均P<0.05);实验药物高剂量组TGF-β_1、PDGF表达水平显著降低(与模型组比较分别P<0.01,P<0.05),实验药物低剂量组无明显作用(P>0.05)。结论蒙古山萝卜酸具有防治实验性大鼠肝损伤纤维化,与MMP-1、TIMP-1酶活性及TGF-β_1、PDGF表达水平失衡的调节作用有关。
Objective To investigate the effect of Mongolia scabiosa acid(MSA)on serum MMP-1,TIMP-1and liver expression level of TGF-β_1,PDGF in model rats with experimental liver fibrosis.Methods Rats were divided into control group,model group,positive control group(colchicine 1.5mg·kg-1)and test drug high dose group,low dose group(MSA 240 and 30mg·kg-1),10 in each group.Except for the control group,250mL·L~(-1) carbon tetrachloride peanut oil was injected in hypodermic,consecutive injection of 4d,a total of 30 times.After injections,the positive control group,high and low dose group began to give MSA,ig,1/d.After 30 times,the blood samples were collected from the rats abdominal aortic,and serum was separated and determined by ELISA.The isolated liver tissue was measured by immunohistochemistry.Results MMP-1and TIMP-1expressed certainly in the control group.MMP-1expression level was obviously reduced in the model group compared with the control group(P<0.05),while TIMP-1expression level was significantly higher than the control group(P<0.05).MMP-1expression showed no obvious change in positive medicine group when compared with the model group(P>0.05).TIMP-1significantly reduced when compared with the model group(P<0.05).MMP-1expression level significantly increased in experimental drug high-dose group when compared with the model group(P<0.05).TIMP-1expression level decreased significantly(P<0.05),MMP-1/TIMP-1ratio increased.MMP-1and TIMP-1expression level showed no obvious effect in experimental drug low dose group(compared with the model group P>0.05).TGF-β_1and PDGF expression levels were low in the control group.TGF-β_1,PDGF expression level increased significantly in the model group(compared with the control group P<0.01).TGF-β_1and PDGF expression level decreased significantly in the positive medicine group(compared with model group P<0.05).TGF-β_1and PDGF expression level decreased significantly in the experimental drug high-dose group(compared with model group,respectively P<0.01,P<0.05).TGF-β_1and PDGF showed no obvious effect in low dose group(P>0.05).Conclusion MSA has the effect of antiliver fibrosis in rats which involved the mechanism of MMP-1,TIMP-1and TGF-β_1,PDGF imbalance expression level adjustment.
Objective To investigate the effect of Mongolia scabiosa acid(MSA)on serum MMP-1,TIMP-1and liver expression level of TGF-β_1,PDGF in model rats with experimental liver fibrosis.Methods Rats were divided into control group,model group,positive control group(colchicine 1.5mg·kg-1)and test drug high dose group,low dose group(MSA 240 and 30mg·kg-1),10 in each group.Except for the control group,250mL·L~(-1) carbon tetrachloride peanut oil was injected in hypodermic,consecutive injection of 4d,a total of 30 times.After injections,the positive control group,high and low dose group began to give MSA,ig,1/d.After 30 times,the blood samples were collected from the rats abdominal aortic,and serum was separated and determined by ELISA.The isolated liver tissue was measured by immunohistochemistry.Results MMP-1and TIMP-1expressed certainly in the control group.MMP-1expression level was obviously reduced in the model group compared with the control group(P<0.05),while TIMP-1expression level was significantly higher than the control group(P<0.05).MMP-1expression showed no obvious change in positive medicine group when compared with the model group(P>0.05).TIMP-1significantly reduced when compared with the model group(P<0.05).MMP-1expression level significantly increased in experimental drug high-dose group when compared with the model group(P<0.05).TIMP-1expression level decreased significantly(P<0.05),MMP-1/TIMP-1ratio increased.MMP-1and TIMP-1expression level showed no obvious effect in experimental drug low dose group(compared with the model group P>0.05).TGF-β_1and PDGF expression levels were low in the control group.TGF-β_1,PDGF expression level increased significantly in the model group(compared with the control group P<0.01).TGF-β_1and PDGF expression level decreased significantly in the positive medicine group(compared with model group P<0.05).TGF-β_1and PDGF expression level decreased significantly in the experimental drug high-dose group(compared with model group,respectively P<0.01,P<0.05).TGF-β_1and PDGF showed no obvious effect in low dose group(P>0.05).Conclusion MSA has the effect of antiliver fibrosis in rats which involved the mechanism of MMP-1,TIMP-1and TGF-β_1,PDGF imbalance expression level adjustment.
引文
[1]Yang C,Zeisberg M,Mosterman B,et a1.Liver fibrosis:insights into migration of hepatic stellate cells in response to extracellular matrix and growth factors[J].Gastroenterology,2003,124(1):147-159.
[2]Hernandez-Gea V,Friedman S L.Pathogenesis of liver fibrosis[J].Annu Rev Pathol,2011,6:425-456.
[3]Friedman S L.Mechanisms of hepatic fibrogenesis[J].Gastroenterology,2008,134(6):1655-1669.
[4]饶慧瑛,魏来.肝星状细胞的生物学特性及活化调控机制[J].世界华人消化杂志,2005,13(5):671-674.
[5]王彩生,赵志清.肝纤维化发病机制研究进展[J].内蒙古医学杂志,2010,42(3):331-334.
[6]胡云龙,孔丽.TGFβ1,CTGF信号传导通路在小鼠肝纤维化发生中的作用[J].基础医学与临床,2012,32(1):66-70.
[7]朱碧红,张慧芳,陈永平.大鼠肝纤维化组织中TGF-β1的研究[J].中国微生态学杂志,2013,25(2):150-152.
[8]白音夫,周雪梅,周长凤,等.蒙古山萝卜有效成分的分离、鉴定及含量测定[J].西北药学杂志,2014,29(6):564-566.
[9]常亮,杨宏昕,赵小原,等.蒙古山萝卜酸对大鼠肝损伤纤维化防治作用的研究[J].中国民族医药杂志,2012,18(2):44-46.
[10]王乐,李多伟.元宝枫中绿原酸研究进展[J].西北药学杂志,2009,24(3):230-232.
[11]王振常,杨删,黄晶晶,等.柔肝化纤颗粒对CCl4诱导的肝纤维化大鼠肝脏MMP-2、TIMP-2的影响[J].疑难病杂志,2014,13(1):74-76,82.
[12]陈祝英,何萍,黄仁彬,等.乙肝转阴散保护小鼠CCl4急性肝损伤的生化机理[J].中药材,2007,30(2):208-210.
[13]于萍,歩宏,王华,等.免疫组化结果的图像分析与人工计数方法的对比研究[J].生物医学工程学杂志,2003,20(2):288-290.
[14]姜宪辉,叶华,孙雷.MMP-1、TIMP-1在肝纤维化组织中的表达及其病理意义[J].医学信息,2008,21(12):2291-2294.
[15]聂青和,周永兴,谢玉梅.肝硬化病人血清、肝组织中金属蛋白酶组织抑制因子的表达及意义[J].中华医学杂志,2001,81(13):805-807.
[16]过建春,陈群伟,包剑锋,等.转化生长因子β1在肝纤维化发病中的作用[J].国际消化病杂志,2009,29(2):86-88.
[17]徐新保,何振平,梁志清,等.阻断转化生长因子-β1信号传导治疗大鼠实验性肝纤维化[J].中华肝脏病杂志,2004,12(5):263-266.
[18]陈莲香,舒建昌.PDGF与肝纤维化关系的研究新进展[J].胃肠病学和肝病学杂志,2011,20(1):95-98.
[19]陈勇良,李振燕,姚春甫,等.TGF-β1、PDGF-BB与慢性乙型肝炎肝纤维化相关性研究[J].实用肝脏病杂志,2010,13(2):92,100-103.
[20]寇国先,刘冰,龙波,等.慢性乙型肝炎患者血清TGF-β、MMP-1、TIMP-1水平及其与肝纤维化程度的研究[J].现代医药卫生,2008,24(3):330-332.
[2]Hernandez-Gea V,Friedman S L.Pathogenesis of liver fibrosis[J].Annu Rev Pathol,2011,6:425-456.
[3]Friedman S L.Mechanisms of hepatic fibrogenesis[J].Gastroenterology,2008,134(6):1655-1669.
[4]饶慧瑛,魏来.肝星状细胞的生物学特性及活化调控机制[J].世界华人消化杂志,2005,13(5):671-674.
[5]王彩生,赵志清.肝纤维化发病机制研究进展[J].内蒙古医学杂志,2010,42(3):331-334.
[6]胡云龙,孔丽.TGFβ1,CTGF信号传导通路在小鼠肝纤维化发生中的作用[J].基础医学与临床,2012,32(1):66-70.
[7]朱碧红,张慧芳,陈永平.大鼠肝纤维化组织中TGF-β1的研究[J].中国微生态学杂志,2013,25(2):150-152.
[8]白音夫,周雪梅,周长凤,等.蒙古山萝卜有效成分的分离、鉴定及含量测定[J].西北药学杂志,2014,29(6):564-566.
[9]常亮,杨宏昕,赵小原,等.蒙古山萝卜酸对大鼠肝损伤纤维化防治作用的研究[J].中国民族医药杂志,2012,18(2):44-46.
[10]王乐,李多伟.元宝枫中绿原酸研究进展[J].西北药学杂志,2009,24(3):230-232.
[11]王振常,杨删,黄晶晶,等.柔肝化纤颗粒对CCl4诱导的肝纤维化大鼠肝脏MMP-2、TIMP-2的影响[J].疑难病杂志,2014,13(1):74-76,82.
[12]陈祝英,何萍,黄仁彬,等.乙肝转阴散保护小鼠CCl4急性肝损伤的生化机理[J].中药材,2007,30(2):208-210.
[13]于萍,歩宏,王华,等.免疫组化结果的图像分析与人工计数方法的对比研究[J].生物医学工程学杂志,2003,20(2):288-290.
[14]姜宪辉,叶华,孙雷.MMP-1、TIMP-1在肝纤维化组织中的表达及其病理意义[J].医学信息,2008,21(12):2291-2294.
[15]聂青和,周永兴,谢玉梅.肝硬化病人血清、肝组织中金属蛋白酶组织抑制因子的表达及意义[J].中华医学杂志,2001,81(13):805-807.
[16]过建春,陈群伟,包剑锋,等.转化生长因子β1在肝纤维化发病中的作用[J].国际消化病杂志,2009,29(2):86-88.
[17]徐新保,何振平,梁志清,等.阻断转化生长因子-β1信号传导治疗大鼠实验性肝纤维化[J].中华肝脏病杂志,2004,12(5):263-266.
[18]陈莲香,舒建昌.PDGF与肝纤维化关系的研究新进展[J].胃肠病学和肝病学杂志,2011,20(1):95-98.
[19]陈勇良,李振燕,姚春甫,等.TGF-β1、PDGF-BB与慢性乙型肝炎肝纤维化相关性研究[J].实用肝脏病杂志,2010,13(2):92,100-103.
[20]寇国先,刘冰,龙波,等.慢性乙型肝炎患者血清TGF-β、MMP-1、TIMP-1水平及其与肝纤维化程度的研究[J].现代医药卫生,2008,24(3):330-332.