参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响

吕锦珍; 徐拥建; 胡世平; 冯高飞; 黄裕华; 邓远军; 王欣

doi:10.13422/j.cnki.syfjx.20192302

您当前的位置：

首页 >

文章列表页 >

参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响

经典名方 | 更新时间：2021-04-16

- 参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响
- Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats
- 中国实验方剂学杂志 2020年26卷第2期页码：6-12
- 作者机构：
  
  1.北京中医药大学　深圳医院，广东深圳　 518172
  2.暨南大学　中医药学院，广州　 510632
- 作者简介：
  
  [第一作者]　吕锦珍，硕士，主治医师，从事中西医结合防治肝病基础及临床研究，Tel：0755-28338833-6202，E-mail：671111933@qq.com
  *徐拥建，博士，副主任医师，从事中西医结合防治肝病及恶性肿瘤研究，Tel: 0755-28338833-6202, E-mail：himi@163.com
- 基金信息：
  
  北京中医药大学校级科研项目(2018BUCMXJKY013);深圳市龙岗区医疗卫生科技计划项目(20170405190544730)
- DOI：10.13422/j.cnki.syfjx.20192302
  中图分类号： R2-0; R22; R285.5; R289
- 收稿日期：2019-05-22，
  
  网络出版日期：2019-08-21，
  
  纸质出版日期：2020-01-20
- 稿件说明：
移动端阅览
吕锦珍, 徐拥建, 胡世平, 等. 参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响[J]. 中国实验方剂学杂志, 2020,26(2):6-12.

Jin-zhen LYU, Yong-jian XU, Shi-ping HU, et al. Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 6-12.
吕锦珍, 徐拥建, 胡世平, 等. 参苓白术散对NAFLD大鼠肝细胞mTORC1/STAT3信号通路的影响[J]. 中国实验方剂学杂志, 2020,26(2):6-12. DOI： 10.13422/j.cnki.syfjx.20192302.

Jin-zhen LYU, Yong-jian XU, Shi-ping HU, et al. Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(2): 6-12. DOI： 10.13422/j.cnki.syfjx.20192302.

摘要

目的：

观察参苓白术散对高脂饮食饲养的非酒精性脂肪性肝病(NAFLD)大鼠肝细胞哺乳动物雷帕霉素靶蛋白复合体1(mTORC1)/细胞信号转导因子及转录激活因子3(STAT3)信号通路的影响，从炎症角度揭示参苓白术散抗大鼠NAFLD的作用机制。

方法：

取SD大鼠80只，随机分为4组，分别为正常组、模型组、参苓白术散高、低剂量组(30，10 g·kg

-1

)，每组20只。采用高脂饲料喂养大鼠8周，建立NAFLD大鼠模型，各药物干预组大鼠灌服相应剂量的参苓白术散，8周后取血和肝组织样本，全自动生化分析仪检测血清丙氨酸氨基转移酶(ALT)，天门冬氨酸氨基转移酶(AST)，总胆固醇(TC)，甘油三酯(TG)，高密度脂蛋白胆固醇(HDL-C)，低密度脂蛋白胆固醇(LDL-C)定量；对肝组织进行油红O，苏木素-伊红(HE)染色；采用Ⅳ型胶原酶离体循环灌注法分离肝细胞；酶联免疫吸附测定(ELISA)检测肝细胞肿瘤坏死因子(TNF)-

，白细胞介素(IL)-1

，IL-5及IL-6含量变化，实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测大鼠肝细胞mTORC1，STAT3 mRNA及蛋白表达水平。

结果：

与正常组比较，模型组大鼠病理组织学改变表明，肝组织炎症及脂肪蓄积显著，血清ALT，AST，TC，TG及LDL-C含量显著升高，HDL-C显著下降，肝细胞TNF-

，IL-1

，IL-5及IL-6水平显著升高，mTORC1，STAT3 mRNA及蛋白相对表达量显著升高(

0.01)。与模型组比较，参苓白术散高、低剂量组肝组织脂质蓄积明显改善，血清ALT，AST，TC，TG及LDL-C含量明显下降，肝细胞TNF-

，IL-1

，IL-5及IL-6水平明显下降，肝细胞mTORC1，STAT3 mRNA及蛋白相对表达量明显降低；参苓白术散高剂量组在改善脂质蓄积，抑制肝组织炎症反应方面，效果优于参苓白术散低剂量组，mTORC1，STAT3 mRNA及蛋白表达明显降低(

0.05，

0.01)。

结论：

参苓白术散能够改善高脂饮食诱导的NAFLD大鼠脂肪代谢紊乱、减轻肝脏脂质蓄积及炎症反应，其作用机制可能与抑制肝细胞内mTORC1/STAT3通路相关mRNA及蛋白表达有关。

Abstract

Objective:

To observe the effect of Shenling Baizhusan(SBS)on the mammalian target of rapamycin complex 1 (mTORC1)/signal transducers and activators of transcription 3 (STAT3) pathway in liver hepatocyte of nonalcoholic fatty liver disease(NAFLD)rats induced by high fat diet

in order to reveal the mechanism of SBS against rat NAFLD from the perspective of inflammation.

Method:

Totally 80 SD rats were randomly divided into 4 groups

normal control group

model group

high-dose SBP group(30 g·kg

-1

)

and low-dose SBS group(10 g·kg

-1

)

with 20 rats in each group. The rats of NAFLD model were established by being fed with high-fat diets for 8 weeks

and the treatment groups were fed with high or low dose of SBS respectively. After treatment for 8 weeks

blood and liver samples of rats were collected. Alanine aminotransferase (ALT)

aspartate aminotransferase(AST)

total cholesterol (TC)

triglyceride(TG)

high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels in blood serum were detected with automatic biochemical analyzer. The liver tissues were observed by oil red O and hematoxylin-eosin (HE) staining. Hepatocytes were isolated by type Ⅳ collagenase perfusion

in vitro

. Tumor necrosis factor (TNF)-

interleukin (IL)-1

IL-5 and IL-6 in hepatocytes were detected by enzyme-linked immunosorbent assay (ELISA)

and the relevant gene and proteins expressions of mTORC1 and STAT3 in hepatocytes were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot detection respectively.

Result:

Compared with the normal control group

the serum levels of TG

AST

ALT and LDL-C were increased significantly

the levels of TNF-

IL-1

IL-5 and IL-6 in hepatocytes were increased significantly

and the expression levels of mTORC1

STAT3 mRNA and proteins in hepatocytes were increased significantly(

0.01). Compared with the model group

the hepatic lipid accumulation of the medicine intervention group was relieved significantly

the serum levels of AST

ALT

TG and LDL-C were decreased significantly

the expression levels of TNF-

IL-1

IL-5 and IL-6 of hepatocytes were decreased significantly

and the expressions of mTORC1

STAT3 mRNA and proteins in hepatocytes were decreased significantly(

0.05

0.01). In the high-dose SBS group

the effects in improving the lipid accumulation and inhibiting the inflammatory reaction were better than those of the low-dose SBS group

and the expressions of mTORC1 and STAT3 genes and proteins in hepatocytes were significantly decreased (

0.05

0.01).

Conclusion:

SBS can improve the fat metabolism disorder and reduce liver lipid accumulation and inflammatory reaction in NAFLD rats induced by high-fat diet. The mechanism may be correlated with the inhibition of mTORC1/STAT3 pathway relating to genes and protein expression in hepatocytes.

关键词

Keywords

references

J D Browning ， L S Szczepaniak ， R Dobbins ， et al . Prevalence of hepatic steatosis in an urban population in the United States: impact of ethnicity [J]． Hepatology ， 2004 ， 40 ( 6 )： 1387 - 1395 .

M Blachier ， H Leleu ， M Peck-Radosavljevic ， et al . The burden of liver disease in Europe: a review of available epidemiological data [J]． J Hepatol ， 2013 ， 58 : 593 - 608 .

K Q SHI ， Y C FAN ， W Y LIU ， et al . Traditional Chinese medicines benefit to nonalcoholic fatty liver disease: a systematic review and Meta-analysis [J]． Mol Biol Rep ， 2012 ， 39 ( 10 ) : 9715 - 9722 .

刘志华，孙晓波．网络药理学：中医药现代化的新机遇 [J]．药学学报， 2012 ， 47 ( 6 )： 696 - 703 .

张玉佩，杨钦河，孔怡琳，等．从痰瘀角度探讨脂肪肝“二次打击”学说 [J]．新中医， 2010 ， 10 ( 7 )： 158 - 159 .

王珏云，张异卓，邹金桥，等．参苓白术散治疗非酒精性脂肪肝随机对照试验Meta分析 [J]．辽宁中医药大学学报， 2017 ， 19 ( 9 )： 110 - 114 .

Q H YANG ， Y J XU ， G F FENG ， et al . p38 MAPK signal pathway involved in anti-inflammatory effect of chaihu-shugan-san and shen-ling-bai-zhu-san on hepatocyte in non-alcoholic steatohepatitis rats [J]． Afr J Tradit Complem Med ， 2014 ， 11 ( 1 )： 213 - 221 .

徐拥建，杨钦河，韩莉，等．疏肝健脾方药对NAFLD大鼠肝细胞SREBP-1c、SCD-1 mRNA及蛋白表达的影响 [J]．中药材， 2014 ， 37 ( 1 )： 83 - 89 .

S Sciarretta ， M Forte ， G Frati ， et al . New insights into the role of mTOR signaling in the cardiovascular system [J]． Circ Res ， 2018 ， 122 ( 3 )： 489 - 505 .

A Moeini ， H Cornellà ， A Villanueva ． Emerging signaling pathways in hepatocellular carcinoma [J]． Liver Canc ， 2012 ， 1 ( 2 ): 83 - 93 .

L Belloni ， S Di Cocco ， F Guerrieri ， et al . Targeting a phospho-STAT3-miRNAs pathway improves vesicular hepatic steatosis in an in vitro and in vivo model [J]． Sci Rep ， 2018 ， 8 ( 1 )： 13638 .

段富津．方剂学 [M]．上海：上海科学技术出版社， 1995 ： 114 - 179 .

施新猷．现代医学实验动物学 [M]．北京：人民军医出版社， 2000 ： 334 .

中华中医药学会脾胃病分会．非酒精性脂肪性肝病中医诊疗专家共识意见(2017) [J]．中医杂志， 2017 ， 58 ( 19 )： 1707 - 1708 .

丁凌辉，贾育新，成映霞，等．参苓白术散对脾虚湿困型溃疡性结肠炎大鼠结肠IL-13，IL-23及COX-2，CREB表达的影响 [J]．中国实验方剂学杂志， 2018 ， 24 ( 11 )： 67 - 72 .

张广玉，张勤生，孙晓娜，等．参苓白术散加减治疗抗生素相关性腹泻脾胃虚寒证的临床观察 [J]．中国实验方剂学杂志， 2019 ， doi： 10.13422/j.cnki.syfjx.20191835 http://dx.doi.org/10.13422/j.cnki.syfjx.20191835 .

张栎婧，战丽彬．基于整合药理学平台探究参苓白术散治疗2型糖尿病的物质基础和作用机制 [J]．中国实验方剂学杂志， 2018 ， 24 ( 21 )： 157 - 162 .

R A Saxton ， D M Sabatini ． mTOR signaling in growth，metabolism，and disease [J]． Cell ， 2017 ， 169 ( 2 )： 361 - 371 .

S Wullschleger ， R Loewith ， M N Hall ． TOR signaling in growth and metabolism [J]. Cell ， 2006 ， 124 ( 3 )： 471 - 484 .

E Choil ， W Kim ， S K Joo ， et al . Expression patterns of STAT3 ERK and estrogen-receptor α are associated with development and histologic severity of hepatic steatosis: a retrospective study [J]. Diagn Pathol ， 2018 ， 13 ( 1 )： 23 .

S Thiem ， T P Pierce ， M Palmieri ， et al . mTORC1 inhibition restricts inflammation-associated gastrointestinal tumorigenesis in mice [J]. J Clin Invest , 2013 ， 123 ( 2 ): 767 - 781 .

Z HE ， X HE ， Z CHEN ， et al . Activation of the mTORC1 and STAT3 pathways promotes the malignant transformation of colitis in mice [J]. Oncol Rep ， 2014 ， 32 ( 5 ): 1873 - 1880 .

W BAO ， Y WANG ， Y FU ， et al . mTORC1 regulates flagellin-induced inflammatory response in macrophages [J]． PLoS One ， 2015 ， 10 ( 5 )： e0125910 .

C ZHU ， L XIA ， F LI ， et al . mTOR complexes differentially orchestrates eosinophil development in allergy [J]． Sci Rep ， 2018 ， 8 ( 1 )： 6883 .

E D Smith ， G A Prieto ， L TONG ， et al . Rapamycin and interleukin-1 β impair brain-derived neurotrophic factor-dependent neuron survival by modulating autophagy [J]． J Biol Chem ， 2014 ， 289 ( 30 )： 20615 - 20629 .

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制

基于p38 MAPK通路探究参苓白术散对克罗恩病大鼠肠道炎症反应的影响

参苓白术散调节Nrf2/ARE通路改善高尿酸血症型少弱精子症小鼠的生精功能

基于Nrf2信号通路探究参苓白术散调节铁死亡对酒精性肝损伤大鼠的影响

参苓白术散在多种疾病模型中的潜在作用机制