温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-<italic style="font-style: italic">α</italic>及肝细胞凋亡的影响

杜珊; 王杰; 陈斌; 周为; 李武; 苏煦初; 彭杰; 周月

doi:10.13422/j.cnki.syfjx.20200222

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温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-α及肝细胞凋亡的影响

中医药抗肝衰竭炎症反应的免疫学机制研究专题 | 更新时间：2021-04-16

- 温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-α及肝细胞凋亡的影响
- Effect of Wenyang Jiedu Huayu Granule on TLR4 mRNA, TNF-α and Hepatocyte Apoptosis in IETM Rats with Hepatic Failure
- 中国实验方剂学杂志 2020年26卷第2期页码：26-32
- 作者机构：
  
  1.湖南中医药大学，长沙　 410208
  2.金华市中医医院，浙江金华　 321017
  3.湖南中医药大学　第一附属医院，长沙　 410007
- 作者简介：
  
  [第一作者]　杜珊，在读博士，从事中医药防治肝病研究，E-mail: ds19910706@163.com
  *陈斌，博士，主任医师，从事中医药防治肝病研究，E-mail: chenbin0410@126.com
- 基金信息：
  
  国家中医临床研究基地业务建设科研专项(JDZX2012062);湖南省中医药科研计划项目(201205);湖南省科技厅科技计划一般项目(2013SK3105)
- DOI：10.13422/j.cnki.syfjx.20200222
  中图分类号： R22; R242; R2-031;R285.5
- 收稿日期：2019-05-09，
  
  网络出版日期：2019-10-09，
  
  纸质出版日期：2020-01-20
- 稿件说明：
移动端阅览
杜珊, 王杰, 陈斌, 等. 温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-α及肝细胞凋亡的影响[J]. 中国实验方剂学杂志, 2020,26(2):26-32.

Shan DU, Jie WANG, Bin CHEN, et al. Effect of Wenyang Jiedu Huayu Granule on TLR4 mRNA, TNF-α and Hepatocyte Apoptosis in IETM Rats with Hepatic Failure[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 26-32.
杜珊, 王杰, 陈斌, 等. 温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-α及肝细胞凋亡的影响[J]. 中国实验方剂学杂志, 2020,26(2):26-32. DOI： 10.13422/j.cnki.syfjx.20200222.

Shan DU, Jie WANG, Bin CHEN, et al. Effect of Wenyang Jiedu Huayu Granule on TLR4 mRNA, TNF-α and Hepatocyte Apoptosis in IETM Rats with Hepatic Failure[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 26-32. DOI： 10.13422/j.cnki.syfjx.20200222.

摘要

目的：

研究Toll样受体4(TLR4) mRNA及其下游炎性因子肿瘤坏死因子-

(TNF-

)与肝衰竭肠源性内毒素血症(IETM)大鼠肝细胞凋亡的关系，并探索温阳解毒化瘀颗粒对内毒素介导的肝细胞凋亡的调控机制。

方法：

SPF级雄性SD大鼠85只，随机分为正常组，模型组，TLR4单克隆抗体组和温阳解毒化瘀颗粒组(8.925 g·kg

-1

)。采用

-半乳糖胺(

-Gal)腹腔注射建立肝衰竭IETM模型，TLR4单克隆抗体组和温阳解毒化瘀颗粒组在造模前5 d给予温阳解毒化瘀颗粒溶液灌胃，正常组、模型组以等容积蒸馏水代替，直至处死。各组分别在24，48，72 h随机处死大鼠并采集标本。检测24，48，72 h各组时间点血清丙氨酸氨基转移酶(ALT)，天门冬氨酸氨基转移酶(AST)水平，苏木素-伊红(HE)染色观察肝组织病理变化，实时荧光定量聚合酶链式反应(Real-time PCR)检测肝组织TLR4 mRNA表达，酶联免疫吸附测定法(ELISA)检测肝组织TNF-

表达，流式细胞仪检测肝细胞凋亡率。

结果：

与正常组比较，模型组ALT，AST升高，肝组织病理损伤程度明显加重，TLR4 mRNA，TNF-

表达均增高(

0.05，

0.01)，肝细胞凋亡率明显上升(

0.05，

0.01)；与模型组比较，温阳解毒化瘀颗粒组ALT，AST显著降低(

0.01)，肝组织病理损伤程度明显减轻(

0.05)，TLR4 mRNA，TNF-

表达显著下降(

0.01)，肝细胞凋亡率亦显著降低(

0.01)。

结论：

TLR4 mRNA，TNF-

在肝衰竭时与肝细胞凋亡呈正相关，温阳解毒化瘀颗粒能够改善肝衰竭IETM大鼠肝功能，减轻肝细胞损伤，减少肝细胞凋亡，其机制可能与其下调肝脏TLR4 mRNA表达，抑制TNF-

释放，降低肝细胞凋亡率有关。

Abstract

Objective:

To investigate the relationship between Toll-like receptor 4 (TLR4) mRNA and its downstream inflammatory factor-tumor necrosis factor-

(TNF-

) and hepatocyte apoptosis in mice with intestinal endotoxemia (IETM) of hepatic failure

and explore the regulatory mechanism of Wenyang Jiedu Huayu granule on endotoxin-induced hepatocyte apoptosis.

Method:

The 85 SPF male SD rats were randomly divided into normal group

model group

TLR4 monoclonal antibody group and Wenyang Jiedu Huayu granule group.

-galactosamine (

-Gal) intraperitoneal injection was performed to establish the IETM model of hepatic failure. The TLR4 monoclonal antibody group and the Wenyang Jiedu Huayu granule group were given Wenyang Jiedu Huayu granule solution by gavage 5 days before the modeling. The normal group and the model group were given isovolumetric distilled water. Each group was given by gavage until sacrifice. Rats in each group were randomly sacrificed at 24

72 h

respectively

and samples were collected. The levels of serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected at 24

72 h. The pathological changes of liver tissue were observed by hematoxylin and eosin(HE) staining. The expression of TLR4 in liver tissue was detected by real-time fluorescence quantitative PCR (Real-time PCR). The expression of TNF-

in liver tissue was detected by enzyme-linked immunosorbent assay (ELISA). The apoptotic rate of hepatocytes was detected by flow cytometry.

Result:

Compared with the normal group

ALT and AST were increased in model group

while the pathological injury degree of liver tissue was significantly increased. The expressions of TLR4 mRNA and TNF-

were increased (

0.05

0.01)

whereas the apoptosis rate of liver cells was significantly increased (

0.05

0.01). Compared with the model group

ALT and AST were decreased in Wenyang Jiedu Huayu granule group (

0.01)

and the degree of pathological injury of liver tissue was significantly reduced (

0.05). The expressions of TLR4 mRNA and TNF-

were significantly decreased (

0.01)

and the apoptosis rate of liver cells was also reduced (

0.01).

Conclusion:

TLR4 mRNA and TNF-

are positively correlated with hepatocyte apoptosis in liver failure. Wenyang Jiedu Huayu granule can improve liver function

alleviate liver cell injury and reduce liver cell apoptosis in IETM mice with hepatic failure. The mechanism may be related to its ability to down-regulate the expression of liver TLR4 mRNA

inhibit the release of TNF-

and reduce the rate of hepatocyte apoptosis.

关键词

Keywords

references

Y YANG , X B GONG , L G HUANG , et al . Diosmetin exerts anti-oxidative, anti-inflammatory and anti-apoptotic effects to protect against endotoxin-induced acute hepatic failure in mice [J]. Oncotarget , 2017 , 8 ( 19 )： 30723 - 30733 .

Y H NIU ， D L YIN ， H L LIU ， et al . Restoring the Treg cell to Th17 cell ratio may alleviate HBV-related acute-on-chronic liver failure [J]. World J Gastroenterol , 2013 ， 19 ( 26 )： 4146 - 4154 .

L DONG , L YIN , H F QUAN , et al . Hepatoprotective effects of kaempferol-3- O - α -l-arabinopyranosyl-7- O - α -l-rhamnopyranoside on D -galactosamine and lipopolysaccharide caused hepatic failure in mice [J]. Molecules , 2017 , 22 ( 10 )： 1755 - 1768 .

S J Kim , S M Lee . Necrostatin-1 protects against D -galactosamine and lipopolysaccharide-induced hepatic injury by preventing TLR4 and RAGE signaling [J]. Inflammation , 2017 ， 40 ( 6 )： 1912 - 1923 .

L P YAN , X N HU , Q H WU , et al . CQMUH-011, a novel adamantane sulfonamide compound, inhibits lipopolysaccharide-and D -galactosamine-induced fulminant hepatic failure in mice [J]． Int Immunopharmacol , 2017 , 47 ( 4 )： 231 - 243 .

王雅，陈斌，龙远雄，等．基于数据挖掘的慢加急性(亚急性)肝衰竭证素分布特点及用药规律研究 [J]．湖南中医药大学学报， 2017 ， 37 ( 11 )： 1234 - 1237 .

莫小艾，蒋开平，黄清华，等．甘露消毒丹治疗乙肝病毒相关慢加急性肝衰竭湿热证疗效及FibroScan变化的临床观察 [J]．中国实验方剂学杂志， 2018 ， 24 ( 16 )： 170 - 175 .

龙富立，陈小明，谢丽，等．解毒化瘀颗粒对急性肝衰竭大鼠细胞因子及肝组织的影响 [J]．中国实验方剂学杂志， 2016 ， 22 ( 20 )： 101 - 105 .

王轩，陈斌，彭杰，等．温阳解毒化瘀颗粒对肝衰竭内毒素血症大鼠肠道sIgA的影响 [J]．中西医结合肝病杂志， 2013 ， 23 ( 3 )： 157 - 159 .

陈斌，王杰，彭杰，等．温阳解毒化瘀颗粒对肝衰竭肠源性内毒素血症大鼠TLR4、NF- κ B表达的影响 [J]．世界科学技术—中医药现代化， 2013 ， 15 ( 5 )： 818 - 824 .

陈奇，沈映君，孙学蕙，等．中药药理实验 [M]．贵阳：贵州人民出版社， 1988 ： 21 - 23 .

艾陈芳．抗肝衰复方治疗重型肝炎的临床观察及对急性肝衰竭大鼠防护作用的实验研究 [D]．武汉：湖北中医学院， 2007 .

E C Dullah , C M Ongkudon . Current trends in endotoxin detection and analysis of endotoxin-protein interactions [J]. Crit Rev Biotechnol , 2017 ， 37 ( 2 ): 251 - 261 .

M Murphy , Y XIONG , G Pattabiraman , et al . Pellino-1 positively regulates Toll-like receptor (TLR) 2 and TLR4 signaling and is suppressed upon induction of endotoxin tolerance [J]. J Biol Chem , 2015 , 290 ( 31 )： 19218 - 19232 .

李小苹，李三强，王冬梅，等．肿瘤坏死因子- α 在小鼠酒精性肝损伤过程中的表达及意义 [J]．中国临床药理学杂志， 2017 ， 33 ( 19 )： 1912 - 1914 .

李树志，熊壮，刘铁军．基于下法毒消肝清丸对于肝硬化内毒素血症的临床研究 [J]．辽宁中医杂志， 2018 ， 45 ( 9 )： 1870 - 1872 .

张向磊，卢立伟，胡冬青，等．护肠清毒微丸结肠靶向给药阻断瘀毒内蕴证乙型肝炎相关慢加急性肝衰竭肠源性内毒素血症临床观察 [J]．中国实验方剂学杂志， 2017 ， 23 ( 18 )： 191 - 197 .

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温阳解毒化瘀颗粒对肝衰竭IETM大鼠TLR4 mRNA，TNF-α及肝细胞凋亡的影响

Effect of Wenyang Jiedu Huayu Granule on TLR4 mRNA, TNF-α and Hepatocyte Apoptosis in IETM Rats with Hepatic Failure

DOI：10.13422/j.cnki.syfjx.20200222

摘要

Abstract

关键词

Keywords

references