肝苏颗粒对猪血清致免疫性肝纤维化大鼠肝功能和病理损伤的影响
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摘要
目的:探讨肝苏颗粒对猪血清诱导的大鼠免疫性肝纤维化的防治作用和机制。方法:将大鼠随机分为正常对照(A)组,模型对照(B)组,秋水仙碱(C)组和肝苏颗粒高(D)、中(E)、低(F)剂量组,采用猪血清腹腔注射建立大鼠免疫性肝纤维化模型。给药10周后,取血检测大鼠肝功能,取部分肝组织匀浆后用Elisa法测定透明质酸(HA)、层黏蛋白(LN)、三型前胶原(PCⅢ)、基质金属蛋白酶1(MMP-1)和基质金属蛋白酶抑制剂-1(TIMP-1)含量,以及α-平滑肌肌动蛋白(α-SMA)和转化生长因子-β1(TGF-β1)表达,另取部分肝组织行HE染色和Masson染色观察。结果:与B组比较,各给药组血清ALT、AST、TBIL均显著下降,肝组织HA、LN、PCⅢ和TIMP-1水平显著降低(P<0.05,P<0.01);C组及E组α-SMA、TGF-β1水平和F组TGF-β1水平均显著降低(P<0.05,P<0.01);各给药组纤维化程度有不同程度改善。结论:肝苏颗粒对猪血清致大鼠免疫性肝纤维化有明显的治疗作用,可明显改善肝功能和减轻肝脏纤维化病变,机制与减轻肝细胞损伤和抑制纤维病变发展有关。
Objective: To observe the potential preventive effect and possible mechanism of Gansu Granules on hepatic fibrosis rats induced by pig serum. Methods: Healthy SD rats were divided into 6 groups randomly according to weight, which contained control group(A group), pig serum induced model group(B group), colchicine group(C group), and Gansu Granules groups with different doses(D, E, F group). The model of liver fibrosis was established through intraperitoneal injections of pig serum. The general state of rats after given the corresponding medicine was observed. After 10 weeks of administration, the liver function of rats was measured. After taking part of liver homogenate, the contents of hyaluronic acid(HA), laminin(LN), type procollagen(PCⅢ), metalloproteinase-1(MMP-1) and inhibitor of metalloproteinase-1(TIMP-1), as well as the expressions of alpha-smooth muscle actin(α-SMA) and transforming growth factor-β1(TGF-β1) were determined by Elisa method. Pathological histology was observed by HE staining and Masson staining. Results: Compared with B group, the levels of serum ALT, AST and TBIL were significantly increased in the C, D, E, F groups, the levels of HA, LN, PCⅢ and TIMP-1 in liver were significantly increased(P<0.05, P<0.01). The levels of α-SMA and TGF-β1 in the C and E group and the level of TGF-β1 in the F group were significantly lower than those in the B group(P<0.05, P<0.01). The degree of fibrosis was improved in different degrees. Conclusion: Gansu Granules have obvious therapeutic effect on immune hepatic fibrosis induced by pig serum in rats, and can significantly improve liver function and alleviate liver fibrosis. The mechanism is related to alleviating hepatocyte damage and inhibiting the development of fibrosis.
Objective: To observe the potential preventive effect and possible mechanism of Gansu Granules on hepatic fibrosis rats induced by pig serum. Methods: Healthy SD rats were divided into 6 groups randomly according to weight, which contained control group(A group), pig serum induced model group(B group), colchicine group(C group), and Gansu Granules groups with different doses(D, E, F group). The model of liver fibrosis was established through intraperitoneal injections of pig serum. The general state of rats after given the corresponding medicine was observed. After 10 weeks of administration, the liver function of rats was measured. After taking part of liver homogenate, the contents of hyaluronic acid(HA), laminin(LN), type procollagen(PCⅢ), metalloproteinase-1(MMP-1) and inhibitor of metalloproteinase-1(TIMP-1), as well as the expressions of alpha-smooth muscle actin(α-SMA) and transforming growth factor-β1(TGF-β1) were determined by Elisa method. Pathological histology was observed by HE staining and Masson staining. Results: Compared with B group, the levels of serum ALT, AST and TBIL were significantly increased in the C, D, E, F groups, the levels of HA, LN, PCⅢ and TIMP-1 in liver were significantly increased(P<0.05, P<0.01). The levels of α-SMA and TGF-β1 in the C and E group and the level of TGF-β1 in the F group were significantly lower than those in the B group(P<0.05, P<0.01). The degree of fibrosis was improved in different degrees. Conclusion: Gansu Granules have obvious therapeutic effect on immune hepatic fibrosis induced by pig serum in rats, and can significantly improve liver function and alleviate liver fibrosis. The mechanism is related to alleviating hepatocyte damage and inhibiting the development of fibrosis.
引文
[1]中国中西医结合学会肝病专业委员会.肝纤维化中西医结合诊疗指南.中国肝脏病杂志(电子版),2010,2(4):54-59
[2]黄芹,朱晓宁,汪静.肝纤维化发生机制的研究新进展.西南军医,2016,18(3):264-267
[3]冯浩,王智民,董歌扬,等.赶黄草化学成分的研究.中国中药杂志,2001,26(4):44-46
[4]杨素芳.肝苏颗粒联合恩替卡韦对慢性乙型肝炎患者肝纤维化及乙型肝炎病毒-DNA转阴率的影响.新乡医学院学报,2012,29(7):527-528,531
[5]曲颖,宗蕾,沈镭,等.肝苏对CCl4肝纤维化大鼠氧化应激和胶原表达的影响.胃肠病学,2011,16(4):196-201
[6]谢君,谢晓芳,代良萍,等.肝苏颗粒对四氯化碳致肝纤维化大鼠肝功能和病理损伤的影响.中国实验方剂学杂志,2017,23(8):117-123
[7]张红.软肝冲剂对免疫性肝纤维化大鼠的实验研究.沈阳:辽宁中医药大学,2012
[8]Ishak K,Baptista A,Bianchi L,et al.Histological grading and staging of chronic hepatitis.J Hepatol,1995,22(6):696-699
[9]谌辉,张景辉,刘文琪.姜黄素抗血吸虫病肝纤维化及其机制的实验研究.中草药,2010,41(8):1274-1277
[10]Povero D,Busletta C,Novo E,et al.Liver fibrosis:A dynamic and potentially reversible process.Histol Histopathol,2010,25(8):1075-1091
[11]乔靖怡,白明,苗明三.基于肝病临床病症特点的动物模型分析.中华中医药杂志,2017,32(2):582-587
[12]李志钢,杨晋翔,张伟,等.肝纤维化实验动物模型的建立与评价.北京中医药大学学报(中医临床版),2009,16(4):43-46
[13]孙妩弋,魏伟,桂双英,等.芍芪多苷抗大鼠免疫性肝纤维化作用及部分机制.中国药理学通报,2010,26(4):492-497
[14]刘建文.药理实验方法学——新技术与新方法.2版.北京:化学工业出版社,2008:208
[15]邵祥强,肖华胜.肝纤维化发病机制与临床诊断的研究进展.世界华人消化杂志,2011,19(3):268-274
[16]赵治凤,贾秋龙.肝纤维化四项联合检测对410例慢性肝炎诊断的临床意义.中国民康医学,2012,24(7):798-799
[17]Costelloe S J,Theocharidou E,Tsochatzis E,et al.Hepascore and hyaluronic acid as markers of fibrosis in liver disease of mixedaetiology.Eur J Gastroenterol Hepatol,2015,27(3):313-320
[18]Istaces N,Gulbis B.Study of fibro test and hyaluronic acid biological variation in healthy volunteers and comparison of serum hyaluronic acid biological variation between chronic liver diseases of different etiology and fibrotic stage using confidence intervals.Clin Biochem,2015,48(10-11):652-657
[19]任俊杰,刘立新.MMPs/TIMPs介导的纤维消融平衡与肝纤维化的研究新进展.中华消化病与影像杂志(电子版),2016,6(4):175-179
[20]Rave-Frānk M,Malik I A,Christiansen H,et al.Rat model of fractionated(2 Gy/day)60 Gy irradiation of the liver:Long-term effects.Radiat Environ Biophys,2013,52(3):321-338
[21]黄艳,黄成,李俊.肝纤维化病程中Kupffer细胞分泌的细胞因子对肝星状细胞活化增殖、凋亡的调控.中国药理学通报,2010,26(1):9-13
[22]袁发浒,冯金梅,刘锴,等.EGCG对日本血吸虫感染小鼠TIMP-1及α-SMA表达和肝纤维化的影响.中国病原生物学杂志,2016,11(5):428-433
[23]刘洋,宋海燕,舒祥兵,等.降脂颗粒改善蛋氨酸-胆碱缺乏饮食诱导的非酒精性脂肪性肝炎的小鼠肝纤维化.中华中医药杂志,2015,30(9):3321-3325
[2]黄芹,朱晓宁,汪静.肝纤维化发生机制的研究新进展.西南军医,2016,18(3):264-267
[3]冯浩,王智民,董歌扬,等.赶黄草化学成分的研究.中国中药杂志,2001,26(4):44-46
[4]杨素芳.肝苏颗粒联合恩替卡韦对慢性乙型肝炎患者肝纤维化及乙型肝炎病毒-DNA转阴率的影响.新乡医学院学报,2012,29(7):527-528,531
[5]曲颖,宗蕾,沈镭,等.肝苏对CCl4肝纤维化大鼠氧化应激和胶原表达的影响.胃肠病学,2011,16(4):196-201
[6]谢君,谢晓芳,代良萍,等.肝苏颗粒对四氯化碳致肝纤维化大鼠肝功能和病理损伤的影响.中国实验方剂学杂志,2017,23(8):117-123
[7]张红.软肝冲剂对免疫性肝纤维化大鼠的实验研究.沈阳:辽宁中医药大学,2012
[8]Ishak K,Baptista A,Bianchi L,et al.Histological grading and staging of chronic hepatitis.J Hepatol,1995,22(6):696-699
[9]谌辉,张景辉,刘文琪.姜黄素抗血吸虫病肝纤维化及其机制的实验研究.中草药,2010,41(8):1274-1277
[10]Povero D,Busletta C,Novo E,et al.Liver fibrosis:A dynamic and potentially reversible process.Histol Histopathol,2010,25(8):1075-1091
[11]乔靖怡,白明,苗明三.基于肝病临床病症特点的动物模型分析.中华中医药杂志,2017,32(2):582-587
[12]李志钢,杨晋翔,张伟,等.肝纤维化实验动物模型的建立与评价.北京中医药大学学报(中医临床版),2009,16(4):43-46
[13]孙妩弋,魏伟,桂双英,等.芍芪多苷抗大鼠免疫性肝纤维化作用及部分机制.中国药理学通报,2010,26(4):492-497
[14]刘建文.药理实验方法学——新技术与新方法.2版.北京:化学工业出版社,2008:208
[15]邵祥强,肖华胜.肝纤维化发病机制与临床诊断的研究进展.世界华人消化杂志,2011,19(3):268-274
[16]赵治凤,贾秋龙.肝纤维化四项联合检测对410例慢性肝炎诊断的临床意义.中国民康医学,2012,24(7):798-799
[17]Costelloe S J,Theocharidou E,Tsochatzis E,et al.Hepascore and hyaluronic acid as markers of fibrosis in liver disease of mixedaetiology.Eur J Gastroenterol Hepatol,2015,27(3):313-320
[18]Istaces N,Gulbis B.Study of fibro test and hyaluronic acid biological variation in healthy volunteers and comparison of serum hyaluronic acid biological variation between chronic liver diseases of different etiology and fibrotic stage using confidence intervals.Clin Biochem,2015,48(10-11):652-657
[19]任俊杰,刘立新.MMPs/TIMPs介导的纤维消融平衡与肝纤维化的研究新进展.中华消化病与影像杂志(电子版),2016,6(4):175-179
[20]Rave-Frānk M,Malik I A,Christiansen H,et al.Rat model of fractionated(2 Gy/day)60 Gy irradiation of the liver:Long-term effects.Radiat Environ Biophys,2013,52(3):321-338
[21]黄艳,黄成,李俊.肝纤维化病程中Kupffer细胞分泌的细胞因子对肝星状细胞活化增殖、凋亡的调控.中国药理学通报,2010,26(1):9-13
[22]袁发浒,冯金梅,刘锴,等.EGCG对日本血吸虫感染小鼠TIMP-1及α-SMA表达和肝纤维化的影响.中国病原生物学杂志,2016,11(5):428-433
[23]刘洋,宋海燕,舒祥兵,等.降脂颗粒改善蛋氨酸-胆碱缺乏饮食诱导的非酒精性脂肪性肝炎的小鼠肝纤维化.中华中医药杂志,2015,30(9):3321-3325