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
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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,2,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,2,30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,2,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,P,<,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,P,<,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,P,<,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,P,<,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,P,<,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,P,<,0.05)。,结论,2,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,2,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,2,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,2,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, TG, LDL, and HDL, and elevated protein expression levels of LPL, CPT1B, and FABP4 in the liver (,P,<,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(,P,<,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 (,P,<,0.05). Compared with the model group, serum TC, TG, LDL, and HDL levels were decreased in the ,Jianpi Huatan, Formula low-dose group and the ezetimibe group (,P,<,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 (,P,<,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 (,P,<,0.05).,Conclusion,2,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
健脾化痰方非酒精性脂肪性肝炎相关肝细胞癌脂质代谢脂蛋白脂肪酶肉碱棕榈酰基转移酶1脂肪酸结合蛋白质4小鼠
Keywords
Jianpi Huatan Formulanon-alcoholic steatohepatitis-related hepatocellular carcinomalipid metabolismlipoprotein lipasecarnitine palmitoyltransferase 1fatty acid binding protein 4mice
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