益气逐瘀解毒方抗四氯化碳诱导大鼠肝纤维化的作用研究
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摘要
目的:研究益气逐瘀解毒方(YQZYJD)对四氯化碳(CCl_4)诱导的大鼠肝纤维化的治疗作用。方法:将SD雄性大鼠给予CCl_4橄榄油混合液进行皮下注射7周,制备肝纤维化大鼠模型,随机分为YQZYJD高、中、低剂量组、阳性对照组、模型组,给予相应治疗,5周后处死大鼠,检测外周血生化指标ALT、AST和GGT,酶联免疫吸附试验法(ELISA)检测透明质酸(HA)和层黏连蛋白(LN),肝组织HE、MASSON染色分别检测肝组织损伤及肝纤维化程度,免疫组化法分别检测大鼠肝组织中α-SMA及TGF-β1表达水平。结果:与阳性对照组相比,YQZYJD高剂量组GGT、HA和LN水平显著降低(P<0.05),病理组织学检测发现,YQZYJD治疗各组的肝组织结构回复均优于模型组,而高剂量组优于阳性对照组;YQZYJD各剂量组α-SMA表达均较模型组显著降低(P<0.05),YQZYJD高剂量组TGF-β1的表达较阳性对照组显著降低(P<0.01)。结论:YQZYJD可促进CCl_4诱导肝纤维化大鼠模型的肝组织结构回复,过度沉积的ECM降解,抑制HSCs的活化,降低TGF-β1的表达水平,改善CCl_4诱导肝纤维化大鼠的肝纤维化程度。
Objectives:To study the effect of Yiqi Zhuyu Jiedu Granule(YQZYJD) on hepatic fibrosis in a carbon tetrachloride(CCl_4)-induced rat model. Methods: Hepatic fibrosis was induced in male Sprague-Dawley rats by subcutaneous injection of mixture of CCl_4 and Olive oil twice a week for 7 weeks. The animals were randomly allocated into six groups(each group, n=14) as follows: high, middle and low dose groups which were treated with different doses of YQZYJD by gavage, the positive group which was treated with Fuzheng Huayu Capsule, the model group and blank control group which received distilled water. After 5 weeks, ALT, AST and GGT were detected by automatic biochemical analyzer and hyaluronic acid(HA) and Laminin(LN) were detectd by ELISA. After that, the effect of YQZYJD on hepatic fibrosis was evaluated by histomorphologically using hematoxylin and eosin(HE) staining and MASSON staining, and the collagen proportionate area was quantified. In addition, fibrosis-related factors, such as transforming growth factor β1(TGF-β1), α-smooth muscle actin(α-SMA) expression levels were evaluated using immunohistochemistry assays. Results:GGT, HA and LN decreased significantly in the high dose group of YQZYJD than that of the positive group(P<0.05). Histological improvement was observed in hepatic fibrosis after YQZYJD treatment(P<0.01). The level of α-SMA in all YQZYJD groups reduced significantly than that of the model group. TGF-β1 in the high dose group reduced than that of the positive group(P<0.01). Conclusions: YQZYJD inhibited the activation of HSCs and reduced the level of TGF-β1, and then attenuated hepatic fibrosis in rats with CCl_4-induced fibrosis.
Objectives:To study the effect of Yiqi Zhuyu Jiedu Granule(YQZYJD) on hepatic fibrosis in a carbon tetrachloride(CCl_4)-induced rat model. Methods: Hepatic fibrosis was induced in male Sprague-Dawley rats by subcutaneous injection of mixture of CCl_4 and Olive oil twice a week for 7 weeks. The animals were randomly allocated into six groups(each group, n=14) as follows: high, middle and low dose groups which were treated with different doses of YQZYJD by gavage, the positive group which was treated with Fuzheng Huayu Capsule, the model group and blank control group which received distilled water. After 5 weeks, ALT, AST and GGT were detected by automatic biochemical analyzer and hyaluronic acid(HA) and Laminin(LN) were detectd by ELISA. After that, the effect of YQZYJD on hepatic fibrosis was evaluated by histomorphologically using hematoxylin and eosin(HE) staining and MASSON staining, and the collagen proportionate area was quantified. In addition, fibrosis-related factors, such as transforming growth factor β1(TGF-β1), α-smooth muscle actin(α-SMA) expression levels were evaluated using immunohistochemistry assays. Results:GGT, HA and LN decreased significantly in the high dose group of YQZYJD than that of the positive group(P<0.05). Histological improvement was observed in hepatic fibrosis after YQZYJD treatment(P<0.01). The level of α-SMA in all YQZYJD groups reduced significantly than that of the model group. TGF-β1 in the high dose group reduced than that of the positive group(P<0.01). Conclusions: YQZYJD inhibited the activation of HSCs and reduced the level of TGF-β1, and then attenuated hepatic fibrosis in rats with CCl_4-induced fibrosis.
引文
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[13] Pellicoro A, Ramachandran P, Iredale JP, et al. Liver fibrosis and repair: immune regulation of wound healing in a solid organ[J]. Nat Rev Immunol, 2014, 14(3): 181-194.
[14] Friedman SL. Hepatic Fibrosis: Emerging Therapies[J]. Dig Dis, 2015, 33(4): 504-507.
[15] Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story[J]. Gut, 2015, 64(5): 830-841.
[16] Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis[J].Adv Drug Deliv Rev, 2017,121:27-42.
[17] Friedman SL. Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver[J]. Physiol Rev, 2008, 88(1): 125-172.
[18] Friedman SL, Sheppard D, Duffield JS, et al. Therapy for fibrotic diseases: nearing the starting line[J]. Sci Transl Med, 2013, 5(167):167sr1.
[19] Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story[J]. Gut, 2015, 64(5): 830-841.
[20] Heymann F, Tacke F. Immunology in the liver-from homeostasis to disease[J]. Nat Rev Gastroenterol Hepatol, 2016, 13(2): 88-110.
[2] Yoon YJ, Friedman SL, Lee YA. Antifibrotic Therapies: Where Are We Now[J]. Semin Liver Dis, 2016,36(1):87-98.
[3] Hardy T, Mann DA. Epigenetics in liver disease: from biology to therapeutics[J]. Gut, 2016, 65(11):1895-1905.
[4] Weiskirchen R1, Tacke F. Liver Fibrosis: From Pathogenesis to Novel Therapies[J]. Dig Dis, 2016, 34(4):410-422.
[5] Tsochatzis EA, Bosch J, Burroughs AK. Liver cirrhosis[J]. Lancet, 2014, 383(9930):1749-1761.
[6] Rockey DC, Bell PD, Hill JA. Fibrosis-a common pathway to organ injury and failure[J]. N Engl J Med, 2015, 372(12):1138-1149.
[7] 叶放, 薛博瑜, 周珉, 等. 论湿热瘀毒与肝纤维化[J]. 南京中医药大学学报, 2005, 21(6): 346-349.
[8] 吕嫒嫒, 薛博瑜. 关幼波治疗慢性肝病经验[J]. 河南中医, 2013, 33(4): 521-522.
[9] 戴克敏. 姜春华治疗肝病验方[J]. 山西中医, 2007, 23(6): 10-11.
[10] 叶放, 薛博瑜, 吴勉华, 等. 重视对慢性肝炎肝纤维化进程中湿热瘀毒证治研究[J]. 中华中医药学刊, 2007, 25(12): 2477-2480.
[11] Hyemin Kim, Hyeonseon Jeong, Seulgi Han, et al. Hyaluronate and its derivatives for customized biomedical applications[J]. Biomaterials, 2017, 123:155-171.
[12] Su TH,Kao JH,Liu CJ. Molecular mechanism and treatment of viral hepatitis-related liver fibrosis[J]. Int J Mol Sci, 2014,15(6): 10578-10604.
[13] Pellicoro A, Ramachandran P, Iredale JP, et al. Liver fibrosis and repair: immune regulation of wound healing in a solid organ[J]. Nat Rev Immunol, 2014, 14(3): 181-194.
[14] Friedman SL. Hepatic Fibrosis: Emerging Therapies[J]. Dig Dis, 2015, 33(4): 504-507.
[15] Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story[J]. Gut, 2015, 64(5): 830-841.
[16] Higashi T, Friedman SL, Hoshida Y. Hepatic stellate cells as key target in liver fibrosis[J].Adv Drug Deliv Rev, 2017,121:27-42.
[17] Friedman SL. Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver[J]. Physiol Rev, 2008, 88(1): 125-172.
[18] Friedman SL, Sheppard D, Duffield JS, et al. Therapy for fibrotic diseases: nearing the starting line[J]. Sci Transl Med, 2013, 5(167):167sr1.
[19] Lee YA, Wallace MC, Friedman SL. Pathobiology of liver fibrosis: a translational success story[J]. Gut, 2015, 64(5): 830-841.
[20] Heymann F, Tacke F. Immunology in the liver-from homeostasis to disease[J]. Nat Rev Gastroenterol Hepatol, 2016, 13(2): 88-110.