[1]史晓燕,许小凡,宋　亮,等.柔木丹颗粒对四氯化碳诱导小鼠肝纤维化的治疗作用[J].新乡医学院学报,2019,36(1):001-6.[doi:10.7683/xxyxyxb.2019.01.001]
　SHI Xiao-yan,XU Xiao-fan,SONG Liang,et al.Effect of Roumudan granules on liver fibrosis induced by carbon tetrachloride in mice[J].Journal of Xinxiang Medical University,2019,36(1):001-6.[doi:10.7683/xxyxyxb.2019.01.001]
点击复制

柔木丹颗粒对四氯化碳诱导小鼠肝纤维化的治疗作用
分享到：

《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
36
期数:
2019年1
页码:
001-6
栏目:
基础研究
出版日期:
2019-01-05

文章信息/Info

Title:
Effect of Roumudan granules on liver fibrosis induced by carbon tetrachloride in mice
作者:
史晓燕1; 许小凡1; 宋　亮1; 李　倩1; 常占杰2; 席　奇2; 李京涛2
（1.陕西中医药大学医学科研实验中心，陕西　咸阳　712046；2.陕西中医药大学附属医院肝病科，陕西　咸阳　712000）
Author(s):
SHI Xiao-yan1; XU Xiao-fan1; SONG Liang1; LI Qian1; CHANG Zhan-jie2; XI Qi2; LI Jing-tao2
（1.Medical Experiment Center of Shaanxi University of Chinese Medicine，Xianyang 712046，Shaanxi Province，China；2.Department of Liver Disease，Affiliated Hospital of Shaanxi University of Chinese Medicine，Xianyang 712000，Shaanxi Province，China）
关键词:
肝纤维化; 柔木丹颗粒; 四氯化碳; α-平滑肌肌动蛋白; 组织金属蛋白酶抑制剂-1; 转化生长因子-β1
Keywords:
liver fibrosis; Roumudan granules; carbon tetrachloride; α-smooth muscle actin; tissue-inhibitor of metalloproteinase-1; transforming growth factor-β1
分类号:
R657.3
DOI:
10.7683/xxyxyxb.2019.01.001
文献标志码:
A
摘要:
目的　探讨柔木丹颗粒对四氯化碳（CCl4）诱导的小鼠肝纤维化的治疗作用。方法　将30只成年昆明小鼠随机分为对照组、肝纤维化组和柔木丹组，每组10只。肝纤维化组和柔木丹组小鼠腹腔注射含体积分数20% CCl4的橄榄油混合液5 mL·kg-1，对照组小鼠腹腔注射相同体积的橄榄油，每周2次，连续8周；第3周起，柔木丹组小鼠灌胃柔木丹颗粒4 g·kg-1，每日1次，共6周；肝纤维化组和对照组小鼠灌胃相同体积的生理盐水。实验前和末次给药后24 h（实验后）分别称各组小鼠的体质量，观察各组小鼠体质量的变化；并于取材时称小鼠肝质量，计算各组小鼠肝体比；取小鼠肝组织，进行苏木精-伊红染色和Masson染色，观察小鼠肝组织病理学变化；免疫组织化学法检测小鼠肝组织中α-平滑肌肌动蛋白（α-SMA）的定位及表达；实时聚合酶链反应检测小鼠肝组织中α-SMA、Ⅰ型胶原蛋白A1（COL1A1）、组织金属蛋白酶抑制剂1（TIMP-1）及转化生长因子-β1（TGF-β1） mRNA的表达；Western blot法检测小鼠肝组织中α-SMA、TGF-β1蛋白的表达。结果　实验前3组小鼠体质量比较差异无统计学意义（F=2.312，P>0.05）。肝纤维化组和柔木丹组小鼠体质量增加显著小于对照组（P<0.01），柔木丹组小鼠体质量增加显著大于肝纤维化组（P<0.01）。肝纤维化组小鼠肝体比显著大于对照组和柔木丹组（P<0.01），对照组与柔木丹组小鼠肝体比比较差异无统计学意义（P>0.05）。肝纤维化组小鼠肝组织中α-SMA蛋白及mRNA相对表达量显著高于对照组（P<0.01），柔木丹组小鼠肝组织中α-SMA蛋白及mRNA相对表达量显著低于肝纤维化组（P<0.01），柔木丹组与对照组小鼠肝组织中α-SMA蛋白及mRNA相对表达量比较差异无统计学意义（P>0.05）。肝纤维化组小鼠肝组织中COL1A1和TIMP-1 mRNA相对表达量显著高于对照组（P<0.01），柔木丹组小鼠肝组织中COL1A1和TIMP-1 mRNA相对表达量显著低于肝纤维化组（P<0.01），柔木丹组小鼠肝组织中COL1A1和TIMP-1 mRNA相对表达量显著高于对照组（P<0.01）。肝纤维化组小鼠肝组织中TGF-β1蛋白及mRNA相对表达量显著高于对照组（P<0.01），柔木丹组小鼠肝组织中TGF-β1蛋白及mRNA相对表达量显著低于肝纤维化组（P<0.01），柔木丹组小鼠肝组织中TGF-β1蛋白及mRNA相对表达量显著高于对照组（P<0.01）。结论　柔木丹颗粒可通过降低肝组织中α-SMA、COL1A1、TIMP-1及TGF-β1的表达而改善CCl4诱导的小鼠肝纤维化。
Abstract:
Objective　To investigate the therapeutic effect of Roumudan granules on liver fibrosis induced by carbon tetrachloride （CCl4） in mice.Methods　Thirty adult male Kunming mice were randomly divided into control group，liver fibrosis group and Roumudan granules group，with ten mice in each group.The mice in the liver fibrosis group and Roumudan granules group were intraperitoneally injected with olive oil mixture containing 20% CCl4 （5 mL·kg-1） to establish liver fibrosis models，the mice in the control group were intraperitoneally injected with the same volume of olive oil,twice a week for 8 weeks.From the third week，the mice in the Roumudan granules group were given Roumudan granules 4 g·kg-1 by intragastric administration，once a day for 6 weeks.The mice in the liver fibrosis group and control group were given the same volume of physiological saline by intragastric administration.The body mass of mice in each group was weighed before and 24 hours after the last administration，and the changes of body mass of mice in each group were observed.The liver mass of mice was weighed and the ratio of liver mass to body mass of mice in each group was calculated.The liver tissues of mice were taken.The pathological changes of liver tissues of mice were observed by hematoxylin-eosin staining and Mason staining.The localization and expression of α-smooth muscle actin （α-SMA） in liver tissues of mice was detected by immunohistochemical method.The expression of α-SMA，collagen type I A1 （COL1A1），tissue-inhibitor of metalloproteinase-1 （TIMP-1） and transforming growth factor-β1 （TGF-β1） mRNA in liver tissues of mice were detected by real-time polymerase chain reaction.The expression of α-SMA and TGF-β1 protein in liver tissues of mice were detected by western blot.Results　There was no significant difference in the body mass of mice among the three groups before experiment （F=2.312，P>0.05）.The body mass gain of mice in the liver fibrosis group and Roumudan granules group was significantly lower than that in the control group （P<0.01）.The body mass gain of mice in the Roumudan granules group was significantly higher than that in the liver fibrosis group （P<0.01）.The ratio of liver mass to body mass of mice in the liver fibrosis group was significantly higher than that in the control group and Roumudan granules group （P<0.01）.There was no significant difference in the ratio of liver mass to body mass of mice between the control group and the Roumudan granules group （P>0.05）.The relative expression of α-SMA protein and mRNA in liver tissues of mice in the liver fibrosis group was significantly higher than that in the control group （P<0.01）.The relative expression of α-SMA protein and mRNA in liver tissues of mice in the Roumudan granules group was significantly lower than that in the liver fibrosis group （P<0.01）.There was no significant difference in the relative expression of α-SMA protein and mRNA between the Roumudan granules group and the control group （P>0.05）.The relative expression of COL1A1 and TIMP-1 mRNA in liver tissues of mice in the liver fibrosis group were significantly higher than that in the control group （P<0.01）.The relative expression of COL1A1 and TIMP-1 mRNA in liver tissues of mice in the Roumudan granules group were significantly lower than that in the liver fibrosis group （P<0.01）.The relative expression of COL1A1 and TIMP-1 mRNA in liver tissues of mice in the Roumudan granules group were significantly higher than that in the control group （P<0.01）.The relative expression of TGF-β1 protein and mRNA in liver tissues of mice in the liver fibrosis group was significantly higher than that in the control group （P<0.01）.The relative expression of TGF-β1 protein and mRNA in liver tissues of mice in the Roumudan granules group was significantly lower than that in the liver fibrosis group （P<0.01）.The relative expression of TGF-β1 protein and mRNA in liver tissues of mice in the Roumudan granules group was significantly higher than that in the control group （P<0.01）.Conclusion　Roumudan granules can improve CCl4-induced liver fibrosis of mice by reducing the expression of α-SMA，COL1A1，TIMP-1 and TGF-β1 in liver tissues.

参考文献/References:

［1］　DONGIOVANNI P，ROMEO S，VALENTI L.Hepatocellular carcinoma in nonalcoholic fatty liver：role of environmental and genetic factors［J］.World J Gastroenterol，2014，20（36）：12945-12955.
［2］　SUN M，KISSELEVA T.Reversibility of liver fibrosis［J］.Clin Res Hepatol Gastroenterol，2015，39（Suppl 1）：S60-S63.
［3］　郭英君.柔木丹颗粒治疗慢性乙型肝炎肝纤维化的临床研究［D］.咸阳：陕西中医药大学，2007.
［4］　罗宏丽，肖顺林，冯碧敏.抑毒调肝合剂对四氯化碳复合因素所致肝纤维化大鼠的保护作用［J］.医药导报，2017，36（11）：1244-1249.
［5］　PANEBIANCO C，OBEN J A，VINCIGUERRA M，et al.Senescence in hepatic stellate cells as a mechanism of liver fibrosis reversal：a putative synergy between retinoic acid and PPAR-gamma signaling［J］.Clin Exp Med，2016，17（3）：269-280.
［6］　HUANG Y，DENG X，LIANG J.Modulation of hepatic stellate cells and reversibility of hepatic fibrosis［J］.Exp Cell Res，2017，352（2）：420-426.
［7］　叶国荣，舒建昌，何雅军，等.四氯化碳腹腔注射致大鼠肝纤维化机理研究［J］.广东药学院学报，2010，26（2）：175-178.
［8］　SUI M，JIANG X，CHEN J，et al.Magnesium isoglycyrrhizinate ameliorates liver fibrosis and hepatic stellate cell activation by regulating ferroptosis signaling pathway［J］.Biomed Pharmacother，2018，106：125-133.
［9］　XU F，LIU C，ZHOU D，et al.TGF-β/SMAD pathway and its regulation in hepatic fibrosis［J］.J Histochem Cytochem，2016，64（3）：157-167.
［10］　刘梅，张斌.从瘀论治肝纤维化［J］.中国中西医结合消化杂志，2018，26（3）：310-314.
［11］　程亚伟，胡衡，杨华.中医辨治肝纤维化进展综述［J］.山东中医杂志，2017，36（11）：986-988.
［12］　陈文玲，王宪东，陈文慧，等.健脾软肝方对四氯化碳致肝纤维化大鼠α-SMA及其mＲNA表达的影响［J］.世界中医药，2018，13（9）：2261-2267，2271.
［13］　KOO J H，LEE H J，KIM W，et al.Endoplasmic reticulum stress in hepatic stellate cells promotes liver fibrosis via PERK-Mediated degradation of HNRNPA1 and up-regulation of SMAD2［J］.Gastroenterology，2016，150（1）：181-193.
［14］　ZHANG X，HAN X，YIN L，et al.Potent effects of dioscin against liver fibrosis［J］.Sci Rep，2015，5：9713.
［15］　HUANG Y，DENG X，LIANG J.Modulation of hepatic stellate cells and reversibility of hepatic fibrosis［J］.Exp Cell Res，2017，352（2）：420-426.

相似文献/References:

[1]王宪波,乔汉臣 桑雁,乔立新.愈肝汤对慢性乙型肝炎患者脂质过氧化和肝纤维化的影响[J].新乡医学院学报,2000,17(04):242.
[2]乔汉臣.非酒精性脂肪肝病研究进展[J].新乡医学院学报,2006,23(06):630.
[3]刘恒兴,邵锋,王环震,等.β-胡萝卜素对肝纤维化大鼠肝组织α-平滑肌肌动蛋白和转化生长因子-β1表达的影响[J].新乡医学院学报,2008,25(01):005.
[4]张超贤,乔汉臣,杨道坤.三甲益肝冲剂对肝纤维化大鼠肝组织结缔组织生长因子表达的影响[J].新乡医学院学报,2008,25(01):009.
[5]喻红霞,申志扬,郭　琦,等.声速匹配组织量化技术对慢性乙型病毒性肝炎肝纤维化的诊断价值[J].新乡医学院学报,2017,34(6):523.[doi:10.7683/xxyxyxb.2017.06.019]
　YU Hong-xia,SHEN Zhi-yang,GUO Qi,et al.Diagnostic value of sound velocity tissue quantification technique for hepatic fibrosis in chronic viral hepatitis B patients[J].Journal of Xinxiang Medical University,2017,34(1):523.[doi:10.7683/xxyxyxb.2017.06.019]
[6]王彩娥,杨鹿奎,李桂芳,等.沙利度胺对四氯化碳致小鼠肝纤维化的影响[J].新乡医学院学报,2021,38(8):706.[doi:10.7683/xxyxyxb.2021.08.002]
　WANG Caie,YANG Lukui,LI Guifang,et al.Effect of thalidomide on liver fibrosis induced by carbon tetrachloride in mice[J].Journal of Xinxiang Medical University,2021,38(1):706.[doi:10.7683/xxyxyxb.2021.08.002]
[7]向保云,李　娟,杨　梅,等.慢性丙型病毒性肝炎患者血小板水平与肝纤维化的相关性[J].新乡医学院学报,2020,37(9):856.[doi:10.7683/xxyxyxb.2020.09.011]
　XIANG Baoyun,LI Juan,YANG Mei,et al.Correlation between platelet level and liver fibrosis in patients with chronic hepatitis C[J].Journal of Xinxiang Medical University,2020,37(1):856.[doi:10.7683/xxyxyxb.2020.09.011]
[8]吴宙光,王　斌,刘　冬,等.环状RNA 42398 调控转化生长因子-β1/Smad信号通路在大鼠胆道闭锁肝纤维化中的作用机制[J].新乡医学院学报,2020,37(11):1013.[doi:10.7683/xxyxyxb.2020.11.003]
　WU Zhouguang,WANG Bin,LIU Dong,et al.Mechanism of role of circular RNA 42398 regulating the transforming growth factor-β1/smad signaling pathway in liver fibrosis of rats with biliary atresia[J].Journal of Xinxiang Medical University,2020,37(1):1013.[doi:10.7683/xxyxyxb.2020.11.003]
[9]朱建凤.恩替卡韦和复方鳖甲软肝片联合治疗乙型肝炎肝硬化疗效观察[J].新乡医学院学报,2020,37(11):1084.[doi:10.7683/xxyxyxb.2020.11.018]
　ZHU Jianfeng.Effect of entecavir combined with compound Biejia Ruangan tablet in the treatment of hepatitis B cirrhosis[J].Journal of Xinxiang Medical University,2020,37(1):1084.[doi:10.7683/xxyxyxb.2020.11.018]
[10]赵巍峰,王园园,李　赢,等.安络化纤丸和恩替卡韦联合治疗对乙型肝炎肝硬化代偿期患者肝纤维化和炎性因子的影响[J].新乡医学院学报,2019,36(4):319.[doi:10.7683/xxyxyxb.2019.04.005]
　ZHAO Wei-feng,WANG Yuan-yuan,LI Ying,et al.Effect of Anluo Huaxian Wan combined with entecavir on the liver fibrosis and inflammatory factors in patients with compensatory hepatitis B cirrhosis[J].Journal of Xinxiang Medical University,2019,36(1):319.[doi:10.7683/xxyxyxb.2019.04.005]

更新日期/Last Update: 2019-01-05