白芍总苷抗大鼠免疫性肝纤维化作用及其部分机制
摘要
目的：探讨白芍总苷(total glucosides of paeony，TGP)对人白蛋白致大鼠免疫性肝纤维化模型的作用及其部分机制。
方法：建立人白蛋白致大鼠免疫性肝纤维化模型，分光光度法检测血清中丙氨酸氨基转移酶(alanine aminotransferase，ALT)、天门冬氨酸氨基转移酶(aspartateaminotransferase，AST)、一氧化氮(nitric oxide，NO)、脯氨酸肽酶(prolidase，PLD)水平和肝匀浆中羟脯氨酸(hydroxyproline，Hyp)、丙二醛(malondiadehyde，MDA)、超氧化物歧化酶(superoxide dismutase，SOD)、谷胱甘肽过氧化物酶(glutathioneperoxidase，GSH-px)含量，放免法检测血清中透明质酸(hyaluronic acid，HA)和Ⅲ型前胶原(procollagen Ⅲ，PCⅢ)水平，ELISA法检测血清中血清转化生长因子β(transforming growth factor-β，TGF-β)水平。HE染色法对肝脏组织作病理检查。
结果：TGP可明显减轻人白蛋白所致的肝纤维化程度，不仅降低肝匀浆Hyp含量和血清HA和PCⅢ含量，还可减轻肝脏纤维化的病理组织学改变。虽然TGP对人白蛋白致肝纤维化大鼠的肝功能和A／G值变化无明显影响，但TGP能改善肝纤维化大鼠肝脏的氧化状态，减少MDA的产生，提高抗氧化酶的活性，同时发现TGP可减少致肝纤维化细胞因子TGF-β的产生。
结论：TGP具有明显抗大鼠免疫性肝纤维化的作用，其机制可能与清除自由基、提高抗氧化酶活性和减少TGF-β的生成等作用有关。
Objective To study the effects and mechanisms of total glucosides ofpaeony (TGP) on immunological hepatic fibrosis induced by human albumin in rats . Methods The model of immunological hepatic fibrosis induced by human albumin was prepared. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) , prolidase (PLD), nitric oxide(NO) in serum, hydroxyproline(Hyp) content, malondiadehyde(MDA) content, superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) activities in liver homogenate were assayed by spectrophotometry; Serum procollagen type III (PC III) and hyaluronic acid (HA) was determined by radioimmunoassays. The level of transforming growth factor- (TGF- )was determined by ELISA method. Hepatic tissue sections were stained with hematoxylin and eosin and examined under a light microscope. Results: Histological results showed that TGP improved the human albumin -induced alterations in the liver structure, alleviated lobular necrosis and significantly lowered collagen content. The anti-fibrotic effect of TGP also confirmed by decreased serum content of HA and PCIII in TGP-treated group. Moreover, the treatment with TGP effectively reduced the hydroxyproline content in liver homogenate. However, the level of ALT and AST increased in fibrotic rat but had no significance compared with normal control, whereas the ratio of A/G decreased also had no significance. TGP had no effect on level of ALT ,AST and the ratio of A/G Furthermore, TGP treatment significant blocked the increase in MDA and NO, associated with a partially elevation in liver total antioxidant capacity including SOD and GSH-px. Meanwhile, TGP treatment significant decreased the production of TGF- 3 .
Conclusion TGP have beneficial effects on hepatic fibrosis in rats by inhibition of collagen synthesis, decreasing oxidative stress and TGF- production.
方法：建立人白蛋白致大鼠免疫性肝纤维化模型，分光光度法检测血清中丙氨酸氨基转移酶(alanine aminotransferase，ALT)、天门冬氨酸氨基转移酶(aspartateaminotransferase，AST)、一氧化氮(nitric oxide，NO)、脯氨酸肽酶(prolidase，PLD)水平和肝匀浆中羟脯氨酸(hydroxyproline，Hyp)、丙二醛(malondiadehyde，MDA)、超氧化物歧化酶(superoxide dismutase，SOD)、谷胱甘肽过氧化物酶(glutathioneperoxidase，GSH-px)含量，放免法检测血清中透明质酸(hyaluronic acid，HA)和Ⅲ型前胶原(procollagen Ⅲ，PCⅢ)水平，ELISA法检测血清中血清转化生长因子β(transforming growth factor-β，TGF-β)水平。HE染色法对肝脏组织作病理检查。
结果：TGP可明显减轻人白蛋白所致的肝纤维化程度，不仅降低肝匀浆Hyp含量和血清HA和PCⅢ含量，还可减轻肝脏纤维化的病理组织学改变。虽然TGP对人白蛋白致肝纤维化大鼠的肝功能和A／G值变化无明显影响，但TGP能改善肝纤维化大鼠肝脏的氧化状态，减少MDA的产生，提高抗氧化酶的活性，同时发现TGP可减少致肝纤维化细胞因子TGF-β的产生。
结论：TGP具有明显抗大鼠免疫性肝纤维化的作用，其机制可能与清除自由基、提高抗氧化酶活性和减少TGF-β的生成等作用有关。
Objective To study the effects and mechanisms of total glucosides ofpaeony (TGP) on immunological hepatic fibrosis induced by human albumin in rats . Methods The model of immunological hepatic fibrosis induced by human albumin was prepared. The levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST) , prolidase (PLD), nitric oxide(NO) in serum, hydroxyproline(Hyp) content, malondiadehyde(MDA) content, superoxide dismutase(SOD) and glutathione peroxidase(GSH-px) activities in liver homogenate were assayed by spectrophotometry; Serum procollagen type III (PC III) and hyaluronic acid (HA) was determined by radioimmunoassays. The level of transforming growth factor- (TGF- )was determined by ELISA method. Hepatic tissue sections were stained with hematoxylin and eosin and examined under a light microscope. Results: Histological results showed that TGP improved the human albumin -induced alterations in the liver structure, alleviated lobular necrosis and significantly lowered collagen content. The anti-fibrotic effect of TGP also confirmed by decreased serum content of HA and PCIII in TGP-treated group. Moreover, the treatment with TGP effectively reduced the hydroxyproline content in liver homogenate. However, the level of ALT and AST increased in fibrotic rat but had no significance compared with normal control, whereas the ratio of A/G decreased also had no significance. TGP had no effect on level of ALT ,AST and the ratio of A/G Furthermore, TGP treatment significant blocked the increase in MDA and NO, associated with a partially elevation in liver total antioxidant capacity including SOD and GSH-px. Meanwhile, TGP treatment significant decreased the production of TGF- 3 .
Conclusion TGP have beneficial effects on hepatic fibrosis in rats by inhibition of collagen synthesis, decreasing oxidative stress and TGF- production.
引文
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2. Friedman SL. The cellular basis of hepatic fibrosis: mechanisms and treatment strategies. N Engl J Med, 1993;328(25):1828-1835
3. Friedman SL. Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury. J Biol Chem, 2000; 275(4):2247-2250
4. Friedman SL. Liver fibrosis-from bench to bedside. J Hepatol,2003 ;38(S1):S38-S53
5．王吉耀．肝纤维化．见：王吉耀主编．现代肝病治疗——理论与进展．上海：上海医科大学出版社，1999；12：99。
6．尚瑞莲．肝纤维化的病因．见：权启镇，孙自勤，王要军主编．新肝脏病学．济南：山东科学技术出版社，2002；304
7．邱德凯，范竹萍．肝纤维化的发病机制及治疗进展．国外医学·消化系疾病分册，2002；22(4)：195-199
8．黄瑾．肝星状细胞活化的启动、维持和调控．国外医学·消化系疾病分册，2002；22(4)：227-230
9. Mann DA, Smart DE. Transcriptional regulation of hepatic stellate cell activation. Gut, 2002;50(6): 891-896
10. Albanis E, Friedman SL. Hepatic fibrosis. Pathogenesis and principles of therapy. Clin Liver Dis. 2001; 5(2): 315-334
11. Du WD, Zhang YE, Zhai WR, Zhou XM. Dynamic changes of type Ⅰ, Ⅲ and Ⅳ collagen synthesis and distribution of collagen-producing cells in carbon tetrachloride-induced rat liver fibrosis. World J Gastroenterol, 1999; 5 (5): 397～403
12. Knittel T, Schuppan D, Meyer zum Buschenfelde K-H, et al. Differential expression
of collagen type Ⅰ,Ⅲ, and Ⅳ by fatting storing (Ito) cells in vitro. Gastroenterology, 1992; 102(5):1724～35
13. Rombouts K, Kisanga E, Hellemans K, Wielant A, Schuppan D, Geerts A. Effect of HMG-CoA reductase inhibitors on proliferation and protein synthesis by rat hepatic stellate cells. J Hepatol, 2003; 38 (5): 564～72
14. Bonis PA, Friedman SL, Kaplan MM. Is liver fibrosis reversible? N Engl J Med,2001; 344(6):452-454
15．李萍．中医药抗肝纤维化研究进展．湖北中医杂志，2001；23(3)：52-54
16．刘永刚，陈厚昌，蒋毅萍．中草药抗肝纤维化的研究．中药材，2001；24(3)：212-214
17．白云峰，彭康．抗肝纤维化中药研究进展．中药药理与临床，2001；14(4)：48-50
18. Shimizu I, Ma YR, Mizobuchi Y, Liu F, Miura T, Nakai Y, Yasuda M, Shiba M, Horie T, Amagaya S, Kawada N, Hori H, Ito S. Effects of Sho-saiko-to, a Japanese herbal medicine, on hepatic fibrosis in rats. Hepatology, 1999;29(1): 149-60
19．杨利侠，夏慧茹，张菊芳．《傅青主女科》白芍平肝法浅析．山西中医，2000；16(5)：51
20．冯瑞芝，肖培根．白芍．见：中国医学科学院药物研究所编．中药志(1)．北京：人民卫生出版社，1982：182-185
21．张晓燕，王金辉，李铣．白芍的化学成分研究．沈阳药科大学学报，2001；18(1)：30-32
22．张晓燕，李铣．白芍的化学研究进展．沈阳药科大学学报，2002；19(1)：70-73
23．徐叔云，陈敏珠，魏伟等，白芍总苷免疫药理与临床。见：周金黄，刘干中主编：中药药理与临床研究进展(第一册)，北京：中国科学技术出版社，1992：49-59
24．魏伟，刘家骏，刘家琴，等．白芍总苷对乙型肝炎的治疗作用及其前景．中国药理学通报，2000；16(5)：597-598
25．郭浩，魏伟，陈敏珠等．白芍总苷对免疫调节细胞的作用．中国药理学与毒理学
志，1993；7(3)：193-195
26．魏文树，汪文琦，王玉山，等．白芍总苷对免疫应答的调控作用．中国药理学通报，1987；3(3)：148-150
27．周玲玲，魏伟，沈玉先，等．白芍总苷对小鼠系统性红斑狼疮样改变的保护作用．中国药理学通报，2002；18(2)：175-177
28．王华，魏伟，岳莉等．白芍总苷对卡介苗加脂多糖引起的小鼠免疫性肝损伤的保护作用．2004；20(8)(已清样)
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