Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats

Jin-zhen LYU; Yong-jian XU; Shi-ping HU; Gao-fei FENG; Yu-hua HUANG; Yuan-jun DENG; Xin WANG

doi:10.13422/j.cnki.syfjx.20192302

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Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats

Classic Prescriptions | 更新时间：2021-04-16

- Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 2, Pages: 6-12(2020)
- 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：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20192302
  CLC： R2-0; R22; R285.5; R289
- Received：22 May 2019，
  
  Published Online：21 August 2019，
  
  Published：20 January 2020
- 稿件说明：
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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：

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

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