基于LPL/CPT1/FABP4通路探讨健脾化痰方对非酒精性脂肪性肝病“炎癌转化”的抑制作用

邵高眩; 孙陈岑; 季光; 徐汉辰

doi:10.3969/j.issn.1006-2157.2023.12.011

您当前的位置：

首页 >

文章列表页 >

基于LPL/CPT1/FABP4通路探讨健脾化痰方对非酒精性脂肪性肝病“炎癌转化”的抑制作用

实验研究 | 更新时间：2024-01-03

- 基于LPL/CPT1/FABP4通路探讨健脾化痰方对非酒精性脂肪性肝病“炎癌转化”的抑制作用
- Exploration on inhibitory effect of Jianpi Huatan Formula on "inflammation-cancer transformation" in non-alcoholic fatty liver disease based on LPL/CPT1/FABP4 pathway
- 北京中医药大学学报 2023年46卷第12期页码：1706-1715
- 作者机构：
  
  1.上海中医药大学附属龙华医院　上海　201200
  2.上海中医药大学中医脾胃病研究所
  3.上海市炎癌转化病证生物学前沿科学研究中心
- 作者简介：
  
  邵高眩，男，在读硕士生
  #徐汉辰，男，博士，研究员，博士生导师，主要研究方向：中医药防治消化系统疾病的应用基础及临床转化，E-mail：hanson0702@126.com
- 基金信息：
  
  国家自然科学基金项目(82104466);2020年度上海市青年科技启明星(20QA1409300);上海高校青年东方学者(QD2019034)
- DOI：10.3969/j.issn.1006-2157.2023.12.011
  中图分类号： R285.5
- 收稿日期：2023-04-21，
  
  网络出版日期：2023-10-13，
  
  纸质出版日期：2023-12-30
- 稿件说明：
移动端阅览
邵高眩, 孙陈岑, 季光, 等. 基于LPL/CPT1/FABP4通路探讨健脾化痰方对非酒精性脂肪性肝病“炎癌转化”的抑制作用[J]. 北京中医药大学学报, 2023,46(12):1706-1715.

SHAO Gaoxuan, SUN Chencen, JI Guang, et al. Exploration on inhibitory effect of Jianpi Huatan Formula on "inflammation-cancer transformation" in non-alcoholic fatty liver disease based on LPL/CPT1/FABP4 pathway[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(12): 1706-1715.
邵高眩, 孙陈岑, 季光, 等. 基于LPL/CPT1/FABP4通路探讨健脾化痰方对非酒精性脂肪性肝病“炎癌转化”的抑制作用[J]. 北京中医药大学学报, 2023,46(12):1706-1715. DOI： 10.3969/j.issn.1006-2157.2023.12.011.

SHAO Gaoxuan, SUN Chencen, JI Guang, et al. Exploration on inhibitory effect of Jianpi Huatan Formula on "inflammation-cancer transformation" in non-alcoholic fatty liver disease based on LPL/CPT1/FABP4 pathway[J]. Journal of beijing university of traditional chinese medicine, 2023, 46(12): 1706-1715. DOI： 10.3969/j.issn.1006-2157.2023.12.011.

摘要

目的

基于脂蛋白脂肪酶(LPL)/肉碱棕榈酰基转移酶1(CPT1)/脂肪酸结合蛋白质4(FABP4)通路探讨健脾化痰方对非酒精性脂肪性肝病小鼠“炎癌转化”的抑制作用。

方法

30只C57BL/6小鼠按随机数字表法分为对照组、模型组、健脾化痰方低剂量组(19.2 g/kg)、健脾化痰方高剂量组(38.4 g/kg)和依折麦布组(10.0 mg/kg)，每组6只。除对照组外，其他各组小鼠腹腔注射二乙基亚硝胺(25 mg/kg)。4周后，除对照组正常饲养外，其他各组小鼠予高脂、高胆固醇饲料喂养26周，建立非酒精性脂肪性肝炎相关肝细胞癌(NASH-HCC)模型。灌胃给药每日1次，持续26周。比较各组小鼠肝脏肿瘤数量、最大肿瘤表面积。HE染色、油红O染色观察肝脏组织炎症、脂肪变性和气球样变。酶联免疫吸附测定法检测肝脏肿瘤坏死因子-α(TNF-α)含量。生化分析仪检测血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)、高密度脂蛋白(HDL)含量。实时荧光PCR法检测肝脏杆状病毒凋亡抑制蛋白5(Birc5)、淋巴细胞抗原6复合位点D(Ly6d)、CD44抗原(CD44)、增殖标志物Ki-67(Mki67)等肿瘤相关标志物的mRNA表达。蛋白质印迹法检测各组小鼠肝脏LPL、CPT1、FABP4的蛋白表达。

结果

与对照组小鼠相比，模型组小鼠肝脏出现肿瘤，Ly6d、Birc5、CD44、Mki67 mRNA水平升高，肝脏损伤明显，脂肪变性及气球样变严重，肝脏有大量脂滴蓄积，血清ALT、AST及肝脏TNF-α均升高，血清TC、TG、LDL、HDL水平升高，肝脏LPL、CPT1B、FABP4蛋白表达升高(均

0.05)。与模型组小鼠相比，健脾化痰方低剂量组和依折麦布组Birc5 mRNA表达降低(均

0.05)；健脾化痰方低、高剂量组和依折麦布组肝脏损伤不同程度地缓解，表现为脂肪变性和气球样变性的减少、脂滴蓄积的减少及肝脏炎症因子TNF-α的降低(均

0.05)。与模型组比较，健脾化痰方低剂量组、依折麦布组血清TC、TG、LDL、HDL降低(均

0.05)；与健脾化痰方低剂量组比较，依折麦布组血清TC、LDL、HDL降低(均

0.05)。与模型组比较，健脾化痰方低剂量组CPT1B、FABP4的蛋白表达降低，健脾化痰方高剂量组LPL、FABP4的蛋白表达降低(均

0.05)。

结论

健脾化痰方可能通过调控LPL/CPT1/FABP4脂质代谢信号通路，促进TC和TG的分解，改善模型小鼠肝组织脂肪变性及其炎症损伤程度，抑制模型小鼠肿瘤发生发展，从而发挥抗NASH-HCC的作用。

Abstract

Objective

To explore the inhibitory effect of

Jianpi Huatan

Formula on "inflammation-cancer transformation" in a mouse model of non-alcoholic fatty liver disease based on the lipoprotein lipase (LPL)/carnitine palmitoyltransferase 1 (CPT1)/fatty acid binding protein 4 (FABP4) pathway.

Methods

According to the random table method

30 C57BL/6 mice were randomly divided into the control group

model group

Jianpi Huatan

Formula low-dose group (19.2 g/kg)

Jianpi Huatan

Formula high-dose group (38.4 g/kg)

and ezetimibe group (10.0 mg/kg)

six mice in each group. Except for the control group

mice in the other groups were injected intraperitoneally with diethylnitrosamine (25 mg/kg). After 4 weeks

except for the control group

mice in the other groups were fed with a high-fat and high-cholesterol diet for 26 weeks to establish the non-alcoholic steatohepatitis-related hepatocellular carcinoma (NASH-HCC) model. The treatment was administered once daily for 26 weeks. The number of liver tumors and the maximum tumor surface area were compared among the groups. HE staining and oil red O staining were used to observe liver inflammation

steatosis

and ballooning. The content of tumor necrosis factor-α (TNF-α) in liver was determined by ELISA. Serum alanine aminotransferase (ALT)

aspartate aminotransferase (AST)

cholesterol (TC)

triglyceride (TG)

low-density lipoprotein (LDL)

and high-density lipoprotein (HDL) levels were measured by a biochemical analyzer. The mRNA levels of tumor-related markers such as Birc5

Ly6d

CD44

and Mki67 in liver were determined by real-time PCR. The protein expression levels of LPL

CPT1

and FABP4 in the liver of each group of mice were measured by Western blotting.

Results

Compared with the control group

mice in the model group showed tumors in the liver

elevated mRNA levels of Ly6d

Birc5

CD44

and Mki67

obvious liver damage

severe steatosis and ballooning

large amounts of lipid droplet accumulation in the liver

significantly elevated serum ALT

serum AST

and hepatic TNF-α levels

elevated levels of serum TC

LDL

and HDL

and elevated protein expression levels of LPL

CPT1B

and FABP4 in the liver (

0.05). Compared with the model group

the mRNA expression of Birc5 was reduced in the

Jianpi Huatan

Formula low-dose group and the ezetimibe group(

0.05); liver injury was alleviated to varying degrees in the

Jianpi Huatan

Formula low- and high-dose groups and the ezetimibe group

as evidenced by a decrease in steatosis and ballooning

a decrease in lipid droplet accumulation

and a decrease in the hepatic inflammatory factor TNF-α (

0.05). Compared with the model group

serum TC

LDL

and HDL levels were decreased in the

Jianpi Huatan

Formula low-dose group and the ezetimibe group (

0.05); and compared with the

Jianpi Huatan

Formula low-dose group

serum TC

LDL

and HDL levels were significantly decreased in the ezetimibe group (

0.05). Compared with the model group

the protein expression levels of CPT1B and FABP4 were decreased in the

Jianpi Huatan

Formula low-dose group

and the protein expression levels of LPL and FABP4 were decreased in the

Jianpi Huatan

Formula high-dose group (

0.05).

Conclusion

Jianpi Huatan

Formula may exert anti-NASH-HCC effects by regulating the LPL/CPT1/FABP4 lipid metabolism signaling pathway

promoting the breakdown of TC and TG

improving liver tissue steatosis and its inflammatory damage in model mice

and further inhibiting tumor development.

关键词

Keywords

references

CHALASANI N , YOUNOSSI Z , LAVINE JE , et al . The diagnosis and management of nonalcoholic fatty liver disease: practice guidance from the American Association for the Study of Liver Diseases [J]. Hepatology , 2018 , 67 ( 1 ): 328 － 357 .

CHALASANI N , YOUNOSSI Z , LAVINE JE , et al . The diagnosis and management of non-alcoholic fatty liver disease: practice guideline by the American Association for the Study of Liver Diseases, American College of Gastroenterology, and the American Gastroenterological Association [J]. Hepatology , 2012 , 55 ( 6 ): 2005 － 2023 .

NOUREDDIN M , YATES KP , VAUGHN IA , et al . Clinical and histological determinants of nonalcoholic steatohepatitis and advanced fibrosis in elderly patients [J]. Hepatology , 2013 , 58 ( 5 ): 1644 － 1654 .

ARGYROU C , MORIS D , VERNADAKIS S . Hepatocellular carcinoma development in non-alcoholic fatty liver disease and non-alcoholic steatohepatitis. Is it going to be the “Plague” of the 21st century? A literature review focusing on pathogenesis, prevention and treatment [J]. J BUON , 2017 , 22 ( 1 ): 6 － 20 .

余思雨，操颖，朱明哲，等 . 化滞柔肝颗粒减轻非酒精性脂肪性肝炎相关肝纤维化的机制研究 [J]. 上海中医药杂志， 2023 , 57 ( 2 ): 41 － 50 .

程莹，张宁苏，潘琳．原发性肝癌中医证候分布特点研究 [J]. 中国处方药， 2022 ， 20 ( 4 )： 143 － 145 .

NOZAKI S , KUBO M , MATSUZAWA Y , et al . Sensitive non-radioisotopic method for measuring lipoprotein lipase and hepatic triglyceride lipase in post-heparin plasma [J]. Clin Chem , 1984 , 30 ( 5 ): 748 － 751 .

LIN H , PATEL S , AFFLECK VS , et al . Fatty acid oxidation is required for the respiration and proliferation of malignant glioma cells [J]. Neuro Oncol , 2017 , 19 ( 1 ): 43 － 54 .

FURUHASHI M , HOTAMISLIGIL GS . Fatty acid-binding proteins: role in metabolic diseases and potential as drug targets [J]. Nat Rev Drug Discov , 2008 , 7 ( 6 ): 489 － 503 .

HUI XY , LI HY , ZHOU ZG , et al . Adipocyte fatty acid-binding protein modulates inflammatory responses in macrophages through a positive feedback loop involving c-Jun NH2－terminal kinases and activator protein－1 [J]. J Biol Chem , 2010 , 285 ( 14 ): 10273 － 10280 .

BRITTON CH , SCHULTZ RA , ZHANG B , et al . Human liver mitochondrial carnitine palmitoyltransferase I: characterization of its cDNA and chromosomal localization and partial analysis of the gene [J]. Proc Natl Acad Sci U S A , 1995 , 92 ( 6 ): 1984 － 1988 .

WARD PS , THOMPSON CB . Metabolic reprogramming: a cancer hallmark even warburg did not anticipate [J]. Cancer Cell , 2012 , 21 ( 3 ): 297 － 308 .

CONDE DE LA ROSA L , GARCIA-RUIZ C , VALLEJO C , et al . STARD1 promotes NASH-driven HCC by sustaining the generation of bile acids through the alternative mitochondrial pathway [J]. J Hepatol , 2021 , 74 ( 6 ): 1429 － 1441 .

FAN JG , CAO HX . Role of diet and nutritional management in non-alcoholic fatty liver disease [J]. J Gastroenterol Hepatol , 2013 , 28 : 81 － 87 .

WANG D , WEI YR , PAGLIASSOTTI MJ . Saturated fatty acids promote endoplasmic reticulum stress and liver injury in rats with hepatic steatosis [J]. Endocrinology , 2006 , 147 ( 2 ): 943 － 951 .

LUUKKONEN PK , SÄDEVIRTA S , ZHOU Y , et al . Saturated fat is more metabolically harmful for the human liver than unsaturated fat or simple sugars [J]. Diabetes Care , 2018 , 41 ( 8 ): 1732 － 1739 .

THOMPSON KJ , AUSTIN RG , NAZARI SS , et al . Altered fatty acid-binding protein 4 (FABP4) expression and function in human and animal models of hepatocellular carcinoma [J]. Liver Int , 2018 , 38 ( 6 ): 1074 － 1083 .

PRENTICE KJ , SAKSI J , HOTAMISLIGIL GS . Adipokine FABP4 integrates energy stores and counterregulatory metabolic responses [J]. J Lipid Res , 2019 , 60 ( 4 ): 734 － 740 .

YANG HR , DENG QM , NI T , et al . Targeted Inhibition of LPL/FABP4/CPT1 fatty acid metabolic axis can effectively prevent the progression of nonalcoholic steatohepatitis to liver cancer [J]. Int J Biol Sci , 2021 , 17 ( 15 ): 4207 － 4222 .

HOSSEINI H , TEIMOURI M , SHABANI M , et al . Resveratrol alleviates non-alcoholic fatty liver disease through epigenetic modification of the Nrf2 signaling pathway [J/OL]. Int J Biochem Cell Biol , 2020 , 119 : 105667 [ 2023－04－11 ]. https://linkinghub.elsevier.com/retrieve/pii/S1357272519302444 https://linkinghub.elsevier.com/retrieve/pii/S1357272519302444 .

ZHANG YP , DENG YJ , TANG KR , et al . Berberine ameliorates high-fat diet-induced non-alcoholic fatty liver disease in rats via activation of SIRT3/AMPK/ACC pathway [J]. Curr Med Sci , 2019 , 39 ( 1 ): 37 － 43 .

LEE AH , SCAPA EF , COHEN DE , et al . Regulation of hepatic lipogenesis by the transcription factor XBP1 [J]. Science , 2008 , 320 ( 5882 ): 1492 － 1496 .

张晟安，刘志栋，戴亮，等．苓桂术甘汤的临床应用：1项系统性评价与Meta分析 [J]. 中草药， 2023 , 54 ( 14 ): 4599 － 4614 .

罗天益 . 卫生宝鉴 [M]. 史欣德，点评 . 北京：中国医药科技出版社， 2021 : 179 .

曲骞，吴煜 . 吴煜教授疏肝健脾法治疗肝癌经验探析 [J]. 西部中医药， 2018 , 31 ( 6 ): 40 － 43 .

师梦雅，周爱民 . 王彦田教授应用补土荣木法治疗肝癌及癌前病变经验探析 [J]. 河北中医药学报， 2016 , 31 ( 4 ): 52 － 54 .

ARGUELLO G , BALBOA E , ARRESE M , et al . Recent insights on the role of cholesterol in non-alcoholic fatty liver disease [J]. Biochim Biophys Acta , 2015 , 1852 ( 9 ): 1765 － 1778 .

BOUTARI C , KARAGIANNIS A , ATHYROS VG . Rosuvastatin and ezetimibe for the treatment of dyslipidemia and hypercholesterolemia [J]. Expert Rev Cardiovasc Ther , 2021 , 19 ( 7 ): 575 － 580 .

浏览量

1765

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

探讨温和灸通过调控AMPK/PGC-1α信号抑制氧化应激反应保护早发性卵巢功能不全模型小鼠卵巢功能的作用

基于肠－脑轴探讨“疏肝调神”针刺对抑郁障碍小鼠肠道菌群、脂多糖和海马TLR4/NF-κB信号通路的影响

补中益气汤调控自身免疫性甲状腺炎小鼠巨噬细胞极化的机制研究

心阳片对慢性心力衰竭小鼠心肌细胞铁死亡的作用及机制研究

基于“虚－瘀－毒”病机的射血分数保留的心力衰竭病证结合动物模型的建立及评价