健脾化瘀祛痰法对apoA-Ⅰ/AMPK/CPT1A信号通路介导的脂肪酸β氧化改善血脂异常的调控作用及其机制

doi:10.13481/j.1671-587X.20220604

Abstract

Abstract:

Objective To explore the effect of the Jianpi Huayu Qutan method on the dyslipidemia in the rats and to elucidate its molecular mechanism based on apolipoprotein A-Ⅰ（apoA-Ⅰ）/ adenosine monophosphate activated protein kinase （AMPK） / carnitine palmitoyltransferase Ⅰ A（CPT1A） signaling pathway. Methods A total of 32 rats were randomly divided into blank control group， model group， simvastatin group （given 1.575 mg·kg^－1·d^－1 simvastatin） and Huayu Qutan Prescription（HYQTP） group （given 13.846 mg·kg^－1·d^－1 HYQTP drug）， and there were 8 rats in each group. The rats in blank control group were given normal diet，and the rats in model group， simvastatin group and HYQTP group were given high-fat diet to establish the hyperlipidemia rat models. After 8 weeks， the rats in simvastatin group and HYQTP group were given drugs by gavage； the rats in blank control group and model group were given the same volume of normal saline. The levels of total cholesterol （TC）， triglycerides （TG）， low density lipoprotein cholesterol （LDL-c） and high density lipoprotein cholesterol （HDL-c） in serum of the rats in various groups were detected by automatic biochemical analyzer；HE staining was used to observe the pathomorphology of liver tissue of the rats in various groups； Oil red O staining was used to observe the lipid deposition in liver tissue of the rats in various group； the levels of TG in liver tissue of the rats in various groups were detected；ELISA method was used to detect the levels of apoA-Ⅰ in liver tissue of the rats in various groups；real-time fluorescence quantitative PCR （RT-qPCR） method was used to detect the expression levels of apoA-Ⅰ， scavenger receptor class B type 1 （SR-B1）， and CPT1A mRNA in liver tissue of the rats in various groups；Western blotting method was used to detect the expression levels of SR-B1， phosphorylated AMPK （p-AMPK）and CPT1A proteins in liver tissue of the rats in various groups. Results Compared with blank control group， the levels of serum TC， TG，and LDL-c of the rats in model group were increased （P<0.05）， and the level of serum HDL-c was decreased （P<0.05）； the level of TG in liver tissue of the rats was increased （P<0.05）； the hepatocytes showed obvious swelling and increased volume， and the obvious red lipid droplet distribution could be seen； the expression levels of apoA-Ⅰ，SR-B1， and CPT1A mRNA and protein were decreased （P<0.05）， and the ratio of p-AMPK/AMPK was decreased （P<0.05）；compared with model group， the levels of serum TC， TG，and LDL-c in liver tissue of the rats in simvastatin group and HYQTP group were decreased （P<0.05）， and the level of HDL-c was increased （P<0.05），the level of TG in liver tissue was decreased （P<0.05）；the swelling of hepatocytes was obviously reduced，and the distribution of lipid droplets was decreased obviously； the expression levels of apoA-Ⅰ， SR-B1， and CPT1A mRNA and proteins were increased （P<0.05）， and the ratio of p-AMPK/AMPK was increased （P<0.05）. Conclusion The Jianpi Huayu Qutan method can improve the dyslipidemia and reduce the liver damage and lipid deposition of the hyperlipidemia rats， and its mechanism may be related to regulating the apoA-Ⅰ/AMPK/CPT1A signaling pathway and promoting fatty acid β oxidation in the rats.

Key words: Dyslipidemia, Fatty acid β oxidation, Apolipoprotein A-Ⅰ, Adenosine monophosphate activated protein kinase, Carnitine palmitoy ltransferase ⅠA

CLC Number:

R255.7

Qi ZHANG,Qiuyu ZHAO,Guoyuan SUI,Huihui LIU,Yarong ZHAI,Ning YU,Jie WANG,Xueying QIU,Jiawei MENG,Lianqun JIA. Regulatory effect of Jianpi Huayu Qutan method on dyslipidemia improved by fatty acid β oxidation mediated by apoA-Ⅰ/ AMPK/CPT1A signaling pathway and its mechanism[J].Journal of Jilin University(Medicine Edition), 2022, 48(6): 1395-1402.

Figures/Tables 8

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References 23

1	《中国心血管健康与疾病报告2020》编写组. 《中国心血管健康与疾病报告2020》概述［J］. 中国心血管病研究， 2021， 19（7）： 582-590.
2	JAYARAMAN R， REINIER K， NAIR S， et al. Risk factors of sudden cardiac death in the young： multiple-year community-wide assessment［J］. Circulation， 2018， 137（15）： 1561-1570.
3	诸骏仁，高润霖，赵水平，等. 中国成人血脂异常防治指南（2016年修订版）［J］.中国循环杂志，2016，31（10）： 937-953.
4	魏苏宁，苏雪莹，徐国恒. 肝细胞甘油三酯代谢途径异常与脂肪肝［J］. 中国生物化学与分子生物学报， 2016， 32（2）： 123-132.
5	CANTÓ C， AUWERX J. AMP-activated protein kinase and its downstream transcriptional pathways［J］. Cell Mol Life Sci， 2010， 67（20）： 3407-3423.
6	熊文生，李任先，刘国普，刘友章. 辨治高脂血症重在理脾化痰［J］. 新中医， 1996， 28（11）： 4-5.
7	贾连群，宋囡，张妮，等. 基于“脾主运化”理论探讨肠道微生物稳态与膏脂转输的关系［J］. 中医杂志， 2017， 58（18）： 1554-1557.
8	陈阳，张妮，贾连群. 基于PCR array技术探讨化瘀祛痰方对高脂血症大鼠肝脏脂质代谢信号通路相关基因mRNA表达的影响［J］. 时珍国医国药， 2016，27（4）： 809-811.
9	安冬青，吴宗贵，梁春，等. 血脂异常中西医结合诊疗专家共识［J］. 中国全科医学， 2017， 20（3）： 262-269.
10	郑贵力，王煦，王绵之. 王绵之教授治疗高脂血症学术思想及经验［J］. 北京中医药大学学报，2000， 23（2）： 48-50.
11	孟嘉伟，陈丝，王群，等. 化瘀祛痰方调控长链非编码RNA-NEAT1/miR-27b对高脂血症大鼠胆固醇代谢的影响及机制研究［J］. 中华中医药学刊， 2021， 39（9）： 125-129， 263.
12	BONNEFONT J P， DJOUADI F， PRIP-BUUS C，et al.Carnitine palmitoyltransferases 1 and 2： biochemical， molecular and medical aspects［J］. Mol Aspects Med， 2004， 25（5/6）： 495-520.
13	DU Q Q， TAN Z Q， SHI F， et al. PGC1α/CEBPB/CPT1A axis promotes radiation resistance of nasopharyngeal carcinoma through activating fatty acid oxidation［J］. Cancer Sci， 2019， 110（6）： 2050-2062.
14	KIM M K， KIM S H， YU H S， et al. The effect of clozapine on the AMPK-ACC-CPT1 pathway in the rat frontal cortex［J］. Int J Neuropsychopharmacol， 2012， 15（7）： 907-917.
15	JÄGER S， HANDSCHIN C， ST-PIERRE J， et al. AMP-activated protein kinase （AMPK） action in skeletal muscle via direct phosphorylation of PGC-1alpha［J］. Proc Natl Acad Sci U S A， 2007， 104（29）： 12017-12022.
16	郝梦娇，党院霞，周欣欣，等. 黄芪散对高脂血症大鼠AMPK/ACC/CPT1通路的影响［J］. 中国中药杂志， 2018， 43（12）： 2586-2592.
17	WOODS A， DICKERSON K， HEATH R， et al. Ca²⁺/calmodulin-dependent protein kinase kinase-β acts upstream of AMP-activated protein kinase in mammalian cells［J］. Cell Metab， 2005， 2（1）： 21-33.
18	VIOLLET B， ATHEA Y， MOUNIER R，et al. AMPK：lessons from transgenic and knockout animals［J］. Front Biosci （Landmark Ed）， 2009， 14（1）： 19-44.
19	DREW B G， DUFFY S J， FORMOSA M F， et al. High-density lipoprotein modulates glucose metabolism in patients with type 2 diabetes mellitus［J］. Circulation， 2009， 119（15）： 2103-2111.
20	BOUDABA N， MARION A， HUET C， et al. AMPK re-activation suppresses hepatic steatosis but its downregulation does not promote fatty liver development［J］. EBioMedicine， 2018， 28： 194-209.
21	RAO X， WANG Y T. Apolipoprotein A-I improves hepatic autophagy through the AMPK pathway［J］. Biochimie， 2019， 165： 210-218.
22	COCHRAN B J， ONG K L， MANANDHAR B， et al. APOA1： a protein with multiple therapeutic functions［J］. Curr Atheroscler Rep， 2021， 23（3）： 11.
23	SONG P， KWON Y， YEA K， et al. Apolipoprotein a1 increases mitochondrial biogenesis through AMP-activated protein kinase［J］. Cell Signal， 2015， 27（9）： 1873-1881.

Gene	Forward	Reverse
ApoA-Ⅰ	5'-CGAGCTTCACAAAAACGCGA-3'	5'-CATGCGGTCTCGAAACTCCT-3'
SR-B1	5'-CCTTCGTTTCCAGCCAGACA-3'	5'-CCCACAGGATCTCACCAACC-3'
CPT1A	5'-TCAAACCCATTCGTCTTCTGG-3'	5'-CTGCTTATTTTTCCCTCGTGC-3'

Group	TG	TC	LDL-c	HDL-c
Blank control	0.70±0.22	1.40±0.16	0.37±0.12	0.69±0.06
Model	1.67±0.03^*	2.08±0.42^*	0.88±0.32^*	0.37±0.01^*
Simvastatin	0.88±0.36^?	1.41±0.07^△	0.39±0.12^△	0.54±0.02^△
HYQTP	1.10±0.05^?	1.25±0.19^△	0.24±0.06^△	0.56±0.11^△

Group	TG [m_B/(mmol·g^－1)]	apoA-Ⅰ[ρ_B/(ng·L^－1)]
Blank control	0.009±0.001	8.166±0.223
Model	0.017±0.001^*	7.006±0.170^*
Simvastatin	0.011±0.002^△	7.930±0.454^△
HYQTP	0.013±0.002^△	7.747±0.183^△

Group	apoA-Ⅰ	SR-B1	CPT1A
Blank control	1.000±0.000	1.000±0.000	1.000±0.000
Model	0.540±0.141^*	0.484±0.083^*	0.488±0.010^*
Simvastatin	0.987±0.244^△	1.048±0.325^△	1.017±0.279^△
HYQTP	0.945±0.127^△	0.984±0.178^△	0.999±0.268^△

Group	SR-B1	CPT1A	p-AMPK/AMPK
Blank control	0.998±0.006	1.010±0.030	0.989±0.018
Model	0.743±0.025^*	0.746±0.055^*	0.725±0.015^*
Simvastatin	0.900±0.026^△	0.907±0.032^△	0.857±0.024^△
HYQTP	0.830±0.026^△#	0.839±0.007^△	0.896±0.040^△

Regulatory effect of Jianpi Huayu Qutan method on dyslipidemia improved by fatty acid β oxidation mediated by apoA-Ⅰ/ AMPK/CPT1A signaling pathway and its mechanism

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