低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究

doi:10.12092/j.issn.1009-2501.2019.05.005

中国临床药理学与治疗学 ›› 2019, Vol. 24 ›› Issue (5): 511-516.doi: 10.12092/j.issn.1009-2501.2019.05.005

低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究

郑琼娜¹，武俏俏¹，马哲乐¹，张贤菲¹，干方敏¹，孔庆名²

¹温州医科大学附属乐清医院感染科，乐清 325600，浙江; ²浙江省医学科学院，杭州 310013，浙江

收稿日期:2018-12-14 修回日期:2019-05-09 出版日期:2019-05-26 发布日期:2019-05-28
作者简介:郑琼娜，女，主治医师，本科，研究方向：感染性疾病。 E-mail:zhengliiq@126.com
基金资助:
浙江省医药卫生科技计划项目（2015KYA059）

Interventional effect of low molecular weight heparin on mice with experimental hepatic fibrosis

ZHENG Qiongna¹, WU Qiaoqiao¹, MA Zhele¹, ZHANG Xianfei¹, GAN Fangmin¹, KONG Qingming²

¹ Department of Infectious, Yueqing Hospital Affiliated to Wenzhou Medical University, Yueqing 325600，Zhejiang, China; ² Zhejiang Academy of Medical Sciences，Hangzhou 310013, Zhejiang, China

Received:2018-12-14 Revised:2019-05-09 Online:2019-05-26 Published:2019-05-28

摘要/Abstract

摘要：

目的:探讨低分子肝素钠（LMWH）对四氯化碳（CCl4）诱导的小鼠肝纤维化的干预作用。方法:将30只KM小鼠随机均分为空白对照组（腹腔注射橄榄油8周，n=10）、模型对照组（CCl4腹腔注射8周，n=10）和LMWH治疗组（CCl4腹腔注射+LMWH皮下注射8周，n=10）。8周后处死所有小鼠，收集血清及肝组织样本。检测血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、总胆红素（TBIL）、直接胆红素（DBIL）和透明质酸（HA）水平；行HE染色、Masson三色染色和PSR染色观察肝组织病理形态及胶原纤维沉积情况；免疫组化检测肝组织α-平滑肌肌动蛋白（α-SMA）表达量；实时聚合酶链反应（RT-PCR）检测小鼠肝组织中纤维相关因子α-SMA和Ⅰ型胶原（COL-Ⅰ）mRNA表达水平。结果:与空白对照组比较，8周后模型对照组和LMWH治疗组小鼠血清ALT、AST、TBIL、DBIL和HA水平明显升高，肝组织胶原纤维沉积增多，肝组织α-SMA表达增多，α-SMA 和COL-ⅠmRNA表达水平显著升高（均P＜0.05）。与模型对照组比较，8周后LMWH治疗组小鼠血清ALT、AST、TBIL、DBIL和HA水平降低，肝组织胶原纤维沉积减少，肝组织α-SMA表达减少，肝组织α-SMA 和COL-ⅠmRNA表达降低（均P＜0.05）。结论:LMWH可抑制CCl4诱导的小鼠肝脏损害及纤维化，其机制可能与LMWH的抗凝作用有关。

关键词: 低分子肝素钠, 肝纤维化, 动物模型, α-平滑肌肌动蛋白

Abstract:

AIM: To investigate the interventional effect of low molecular weight heparin on mice with hepatic fibrosis caused by carbon tetrachloride (CCl4). METHODS: Thirty mice were randomly divided into three groups: normal control group (intraperitoneally injection of olive oil for 8 weeks, n=10), model group (intraperitoneally injection of CCl4 for 8 weeks, n=10) and interventional group (intraperitoneally injection of CCl4 and subcutaneous injection of low molecular weight heparin for 8 weeks, n=10). All animals were sacrificed at the end of week 8, and blood samples and liver tissue samples were obtained. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL) and hyaluronicacid (HA) were detected. Liver pathology morphology and collagen deposition were observed by HE staining, masson-trichrome staining and PCR staining. The expression of α-smooth muscle actin (α-SMA) in hepatic tissue was detected by immunohistochemistry. The expression of α-SMA and collagen type Ⅰ(COL-Ⅰ) mRNA in the hepatic tissue was quantified by real time PCR. RESULTS:Compared with control group, the serum level of ALT, AST, TBIL, DBIL and HA, the liver collagen deposition, the hepatic tissue expression of α-SMA and the hepatic tissue expression of α-SMA and COL-Ⅰ mRNA in both model group and interventional group were all significantly higher(all P＜0.05). Compared with model group, the serum level of ALT, AST, TBIL, DBIL and HA, the liver collagen deposition, the hepatic tissue expression of α-SMA and the hepatic tissue expression of α-SMA and COL-Ⅰ mRNA in interventional group were all significantly decreased (all P＜0.05).CONCLUSION: Low molecular weight heparin can attenuate the progression of mice hepatic damage and fibrosis caused by CCl4. The mechanism may be related with the anticoagulation of low molecular weight heparin.

Key words: low molecular weight heparin, liver fibrosis, animal model, α-smooth muscle actin

中图分类号:

R332
R575.2

郑琼娜，武俏俏，马哲乐，张贤菲，干方敏，孔庆名. 低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究[J]. 中国临床药理学与治疗学, 2019, 24(5): 511-516.

ZHENG Qiongna, WU Qiaoqiao, MA Zhele, ZHANG Xianfei, GAN Fangmin, KONG Qingming. Interventional effect of low molecular weight heparin on mice with experimental hepatic fibrosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(5): 511-516.

参考文献

［1］Chahal HS, Marseille EA, Tice JA, et al. Cost-effectiveness of early treatment of hepatitis C virus genotype 1 by stage of liver fibrosis in a US treatment-naive population［J］. JAMA Intern Med, 2016, 176(1):65-73.
［2］Hakuno D, Kimura M, Ito SHepatokine, et al. α1-Microglobulin signaling exacerbates inflammation and disturbs fibrotic repair in mouse myocardial infarction［J］. Sci Rep, 2018, 8(1):16749.
［3］Nakao T, Arii S, Kaido T, et al. Heparin accclcrates liver regeneration following portal branch ligation in normal and cirrhotic rats with increased plasma hepatocyte growth factor levels［J］. J Hepatol, 2002,37(1):87-92.
［4］Yan Y, Guan C, Du S, et al. Effects of enzymatically depolymerized low molecular weight heparins on CCl4-induced liver fibrosis［J］. Front Pharmacol, 2017, 8:514.
［5］Tripodi A, Primignani M, Braham S, et al. Coagulation parameters in patients with cirrhosis and portal vein thrombosis treated sequentially with low molecular weight heparin and vitamin K antagonists［J］. Dig Liver Dis, 2016, 48(10):1208-1213.
［6］Kim WR, Berg T, Asselah T, et al. Evaluation of APRI and FIB-4 scoring systems for non-invasive assessment of hepatic fibrosis in chronic hepatitis B patients ［J］. J Hepatol, 2016, 64(4):773-780.
［7］Hui J, Gao J, Wang Y, et al. Panax notoginseng saponins ameliorates experimental hepatic fibrosis and hepatic stellate cell proliferation by inhibiting the Jak2/Stat3 pathways［J］. J Tradit Chin Med, 2016, 36(2):217-224.
［8］潘旭旺, 童永喜, 庄让笑, 等. 芹菜素纳米乳抗CCl-4诱导大鼠肝纤维化作用实验研究［J］. 中华中医药学刊, 2017, 35(6):1482-1485.
［9］Adatya S, Sunny R, Fitzpatrick MJ, et al. Coagulation factor abnormalities related to discordance between anti-factor Xa and activated partial thromboplastin time in patients supported with continuous-flow left ventricular assist devices.［J］. J Heart Lung Transplant, 2016, 35(11):1311-1320.
［10］Annika W, Victoria A, Debashis H, et al. CD248/endosialin critically regulates hepatic stellate cell proliferation during chronic liver injury via a PDGF-regulated mechanism:［J］. Gut, 2016, 65(7):1175-1185.
［11］Cerini F, Vilaseca M, Lafoz E, et al. Enoxaparin reduces hepatic vascular resistance and portal pressure in cirrhotic rats ［J］. J Hepatol, 2016, 64(4):834-842.
［12］Simonetto DA, Yang H, Yin M, et al. Chronic passive venous congestion drives hepatic fibrogenesis via sinusoidal thrombosis and mechanical forces ［J］. Hepatology, 2015, 61(2):648-659.
［13］Papatheodoridis GV, Papakonstantinou E, Andrioti E, et al. Thrombotic risk factors and extent of liver fibrosis in chronic viral hepatitis ［J］. Gut, 2003, 52(3):404-409.
［14］徐列明. 中医药抗肝纤维化疗效评价的探索［J］. 临床肝胆病杂志, 2017, 33(5):825-828.

[1]	魏凤, 何阳, 樊志强, 刘世坤, 欧阳林旗. 氟非尼酮对二乙基亚硝胺诱导的肝损伤大鼠肝细胞中TGF-β1/Smads通路的抑制作用[J]. 中国临床药理学与治疗学, 2022, 27(7): 739-746.
[2]	陈森, 凡畅, 张家富, 马艳珍, 姜辉. 疏肝健脾方调控TLR4/MyD88/NLRP3信号轴抑制肝纤维化小鼠细胞焦亡的机制研究[J]. 中国临床药理学与治疗学, 2022, 27(10): 1081-1089.
[3]	吴芙蓉, 宁丽娟, 周冉. 小柴胡汤对化学性肝纤维化小鼠的保护作用[J]. 中国临床药理学与治疗学, 2020, 25(5): 481-488.
[4]	曹志文，胡卫华. 生长激素宫腔灌注治疗薄型子宫内膜的研究[J]. 中国临床药理学与治疗学, 2019, 24(9): 983-988.
[5]	曾熹，胡杨柳子，吴亚敏，陈志恒. 欧米伽3脂肪酸对硫代乙酰胺诱导的大鼠肝纤维化的保护作用[J]. 中国临床药理学与治疗学, 2017, 22(8): 892-898.
[6]	刘建刚, 王晓波. 子宫内膜异位症模型大鼠背根神经节神经元中TRPV1、TRPA1表达及其意义[J]. 中国临床药理学与治疗学, 2017, 22(6): 617-621.
[7]	田小霞, 来丽娜, 陈云霞, 李旭炯, 王黎敏, 孟丽, 张丽丽. 甜菜碱对肝硬化大鼠肝脏巨噬细胞极化的影响[J]. 中国临床药理学与治疗学, 2016, 21(9): 961-967.
[8]	曹媛，夏延哲，陈杰，陈孝. 五味子甲素在人肝星状细胞中的抗纤维化作用[J]. 中国临床药理学与治疗学, 2016, 21(8): 878-883.
[9]	田小霞，来丽娜，李旭炯，陈云霞，郭晓姝. 甜菜碱对复合致病因素诱导肝硬化大鼠的作用及机制研究[J]. 中国临床药理学与治疗学, 2016, 21(4): 372-377.
[10]	庄让笑，潘旭旺，方红英，王福根，邵益丹，史婷婷，席建军，潘红英. 芹菜素纳米乳抗大鼠肝纤维化作用及对MMP-1，TIMP-1的影响[J]. 中国临床药理学与治疗学, 2016, 21(10): 1122-1125.
[11]	王玲，黄学宽，万磊，杨玉，刘璐. 复肾功方对慢性肾功能衰竭大鼠肾脏α-平滑肌肌动蛋白表达的影响[J]. 中国临床药理学与治疗学, 2016, 21(1): 33-37.
[12]	方红英, 庄让笑, 席建军, 孙晶晶, 邵益丹, 史婷婷, 潘旭旺, 王福根. 乙酰半胱氨酸活性炭微囊治疗大鼠肝纤维化的实验研究[J]. 中国临床药理学与治疗学, 2015, 20(9): 976-980.
[13]	王福根, 梁伟峰, 席建军, 孙晶晶, 潘金明, 叶荣夏, 潘旭旺, 蔡兆斌. 非酒精性脂肪性肝炎动物模型的建立与应用[J]. 中国临床药理学与治疗学, 2015, 20(7): 835-840.
[14]	张纬建, 许建华, 张洁旻, 陈金梅, 陈秀英, 杨梅春, 吕文龙, 洪金省. 甘草次酸对小鼠急性放射性肺损伤的防治作用[J]. 中国临床药理学与治疗学, 2015, 20(6): 629-633.
[15]	华晓萍, 张莉, 黄利, 谭正怀, 陈庄. 两种实验性肝纤维化大鼠模型肝脏组织学特点的比较研究[J]. 中国临床药理学与治疗学, 2014, 19(7): 747-751.

低分子肝素钠对小鼠实验性肝纤维化干预作用的初步研究

Interventional effect of low molecular weight heparin on mice with experimental hepatic fibrosis

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价