FOLLOWUS
1.Department of Pneumology, Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences, Beijing (100053), China
2.Department of Pulmonary and Critical Care Medicine, Mayo Clinic, Rochester, MN 55905, United States
3.Drug Clinical Trial Agency, Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences, Beijing (100053), China
4.Department of Cardiovascular Medicine, Mayo Clinic, Rochester, MN 55905, United States
5.Department of Gynecology, Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences,Beijing (100053), China
6.Department of Cardiovascular Medicine, Guang'anmen Hospital, Chinese Academy of Chinese Medical Sciences, Beijing (100053), China
Dr. WANG Shi-han, E-mail: wang.shihan@mayo.edu
纸质出版日期:2022-06-01,
网络出版日期:2020-07-04,
录用日期:2019-10-22
Scan for full text
Jing-jing TANG, Guang-xi LI, Zhi-guo LIU, 等. 基于HIF-1α-Angptl4 mRNA信号通路探讨丹蒌片抗低氧诱导血脂异常和动脉硬化的机制[J]. Chinese Journal of Integrative Medicine, 2022,28(6):509-517.
Jing-jing TANG, Guang-xi LI, Zhi-guo LIU, et al. Danlou Tablet Improves Chronic Intermittent Hypoxia-Induced Dyslipidemia and Arteriosclerosis by HIF-1α-Angptl4 mRNA Signaling Pathway[J]. Chinese Journal of Integrative Medicine, 2022,28(6):509-517.
Jing-jing TANG, Guang-xi LI, Zhi-guo LIU, 等. 基于HIF-1α-Angptl4 mRNA信号通路探讨丹蒌片抗低氧诱导血脂异常和动脉硬化的机制[J]. Chinese Journal of Integrative Medicine, 2022,28(6):509-517. DOI: 10.1007/s11655-020-3255-8.
Jing-jing TANG, Guang-xi LI, Zhi-guo LIU, et al. Danlou Tablet Improves Chronic Intermittent Hypoxia-Induced Dyslipidemia and Arteriosclerosis by HIF-1α-Angptl4 mRNA Signaling Pathway[J]. Chinese Journal of Integrative Medicine, 2022,28(6):509-517. DOI: 10.1007/s11655-020-3255-8.
目的:
2
研究丹蒌片 (DLT) 对低氧诱导因子(HIF) 1α-血管生成素样4 (Angptl4) mRNA信号通路的调节作用
探讨其抗慢性间歇性低氧(CIH)诱导的血脂异常与动脉硬化的作用机制.
方法:
2
通过3T3-L1细胞培养成熟脂肪细胞
并将其分为8组
包括对照组(1组和5组
细胞培养级超纯水0.1 mL)、DLT组(2组和6组
1000 μg/mL DLT亚微细米级粉剂溶液 0.1 mL)、二甲基草酰甘氨酸 (DMOG) 组( 3组和7组
DMOG和细胞培养级超纯水0.1 mL)、DMOG + DLT组(4组和8组
DMOG和1000 μg/mL DLT亚微细米级粉剂溶液0.1 mL). 1-4组采用成熟脂肪细胞
5-8组采用HIF-1α-siRNA慢病毒转染的成熟脂肪细胞. 处理24小时后
采用实时聚合酶链反应(RT-PCR)和蛋白质印迹 (Western blot) 法分别检测HIF-1α和Angptl4 mRNA及其蛋白表达水平. 在动物实验中
采用ApoE
-/-
小鼠建立CIH模型. 16只小鼠完全随机分为4组
包括假手术组、CIH模型组[间歇性低氧
生理盐水每日灌胃1次(2 mL/次)]、Angptl4抗体组[间歇性低氧
Angptl4抗体每周腹腔注射1次(30 mg/kg)]
及DLT组[间歇性低氧
DLT每日灌胃一次 (250 mg/kg)]
每组4只. 处理4周后
采用酶联免疫吸附(ELISA)法检测血清总胆固醇(TC)及甘油三酯(TG)表达. 采用苏木精-伊红染色(HE)和CD68染色法观察动脉斑块的形态学特征.
结果:
2
Angptl4的表达与HIF-1α相关
在HIF-1α-siRNA慢病毒转染的成熟脂肪细胞中
Angptl4 mRNA水平下降
但蛋白表达不受DMOG及DLT影响. 成熟脂肪细胞中
在DMOG低氧诱导情况下
DLT可抑制HIF-1α及Angptl4 mRNA水平(
P
<
0.05或
P
<
0.01)
降低HIF-1及Angptl4蛋白的表达(均
P
<
0.01)
该作用在siHIF-1慢病毒转染的情况下也存在(
P
<
0.01). CIH处理后
ApoE
-/-
小鼠TG及TC水平升高 (均
P
<
0.01)
动脉粥样硬化斑块加重. Angptl4抗体及DLT均可降低TG和TC水平 (均
P
<
0.01)
在一定程度上降低动脉粥样硬化斑块面积
缩小动脉壁厚度
减轻动脉粥样硬化病变症状.
结论:
2
DLT可通过HIF-1α-Angptl4 mRNA信号通路抑制Angptl4蛋白水平
从而改善血脂异常和动脉硬化.
Objective:
2
To detect whether Danlou Tablet (DLT) regulates the hypoxia-induced factor (HIF)-1α- angiopoietin-like 4 (Angptl4) mRNA signaling pathway and explore the role of DLT in treating chronic intermittent hypoxia (CIH)-induced dyslipidemia and arteriosclerosis.
Methods:
2
The mature adipocytes were obtained from 3T3-L1 cell culturation and allocated into 8 groups including control groups (Groups 1 and 5
0.1 mL of cell culture grade water); DLT groups (Groups 2 and 6
0.1 mL of 1
000 μg/mL DLT submicron powder solution); dimethyloxalylglycine (DMOG) groups (Groups 3 and 7
DMOG and 0.1 mL of cell culture grade water); DMOG plus DLT groups (Groups 4 and 8
DMOG and 0.1 mL of 1
000 μg/mL DLT submicron powder solution). Groups 1–4 used mature adipocytes and groups 5–8 used HIF-1α-siRNA lentivirus-transfected mature adipocytes. After 24-h treatment
real-time polymerase chain reaction and Western blot were employed to determine the mRNA and protein expression levels of HIF-1α and Angptl4. In animal experiments
the CIH model in ApoE
-/-
mice was established. Sixteen mice were complete randomly divided into 4 groups including sham group
CIH model group [intermittent hypoxia and normal saline (2 mL/time) gavage once a day]
Angptl4 Ab group [intermittent hypoxia and Angptl4 antibody (30 mg/kg) intraperitoneally injected every week]
DLT group [intermittent hypoxia and DLT (250 mg/kg) once a day]
4 mice in each group. After 4-week treatment
enzyme linked immunosorbent assay was used to detect the levels of serum total cholesterol (TC) and triglyceride (TG). Hematoxylin-eosin and CD68 staining were used to observe the morphological properties of arterial plaques.
Results:
2
Angptl4 expression was dependent on HIF-1α
with a reduction in mRNA expression and no response in protein level to DMOG or DLT treatment in relation to siHIF-1α-transfected cells. DLT inhibited HIF-1α and Angptl4 mRNA expression (
P
<
0.05 or
P
<
0.01) and reduced HIF-1α and Angptl4 protein expressions with DMOG in mature adipocytes (all
P
<
0.01)
as the effect on HIF-1α protein also existed in the presence of siHIF-1α (
P
<
0.01). ApoE
-/-
mice treated with CIH had increased TG and TC levels (all
P
<
0.01) and atherosclerotic plaque. Angptl4 antibody and DLT both reduced TG and TC levels (all
P
<
0.01)
as well as reducing atherosclerotic plaque areas
narrowing arterial wall thickness and alleviating atherosclerotic lesion symptoms to some extent.
Conclusion:
2
DLT had positive effects in improving dyslipidemia and arteriosclerosis by inhibiting Angptl4 protein level through HIF-1α-Angptl4 mRNA signaling pathway.
Danlou TabletChinese medicinehypoxia-induced factorangiopoietin-like 4dyslipidemiaarteriosclerosis
Jiang YQ, Xue JS, Xu J, Ji YL, Zhou ZX. Efficacy of continuous positive airway pressure treatment in treating obstructive sleep apnea hypopnea syndrome associated with carotid arteriosclerosis. Exp Ther Med 2017;14:6176-6182.
Ahlin S, Manco M, Panunzi S, Verrastro O, Giannetti G,Prete A, et al. A new sensitive and accurate model to predict moderate to severe obstructive sleep apnea in patients with obesity. Medicine (Baltimore) 2019;98:e16687.
Marin JM, Carrizo SJ, Vicente E, Agusti AG. Long-term cardiovascular outcomes in men with obstructive sleep apnoea-hypopnoea with or without treatment with continuous positive airway pressure: an observational study. Lancet 2005;365:1046-1053.
Arnaud C, Poulain L, Lévy P, Dematteis M. Inflammation contributes to the atherogenic role of intermittent hypoxia in apolipoprotein-E knock out mice. Atherosclerosis 2011;219:425-431.
Jun J, Reinke C, Bedja D, Berkowitz D, Bevans-Fonti S,Li J, et al. Effect of intermittent hypoxia on atherosclerosis in apolipoprotein E-deficient mice. Atherosclerosis 2010;209:381-386.
Arnaud C, Beguin PC, Lantuejoul S, Pepin JL, Guillermet C, Pelli G, et al. The inflammatory preatherosclerotic remodeling induced by intermittent hypoxia is attenuated by RANTES/CCL5 inhibition. Am J Respir Crit Care Med 2011;184:724-731.
Li RC, Bodduluri H, Mathis SP, Kim J, Gozal D. Leukotriene B4 receptor-1 mediates intermittent hypoxia-induced atherogenesis. Am J Respir Crit Care Med 2011;184:124-131.
Drager LF, Bortolotto LA, Lorenzi MC, Figueiredo AC,Krieger EM, Filho GL. Early signs of atherosclerosis in obstructive sleep apnea. Am J Respir Crit Care Med 2005;172:613-618.
Drager LF, Jun J, Polotsky VY. Obstructive sleep apnea and dyslipidemia: implications for atherosclerosis. Curr Opin Endoc Diabet Obes 2010;17:394.
Drager LF, Polotsky VY, Lorenzi-Filho G. Obstructive sleep apnea: an emerging risk factor for atherosclerosis. Chest 2011;140:534-542.
Phillips CL, Yee BJ, Marshall NS, Liu PY, Sullivan DR,Grunstein RR. Continuous positive airway pressure reduces postprandial lipidemia in obstructive sleep apnea. Am J Respir Crit Care Med 2011;184:355-361.
Freiberg J, Tybjaerg-Hansen A, Nordestgaard B. Nonfasting triglycerides and risk of ischemic stroke in the general population. Atheros Suppl 2008;9:15.
Drager LF, Li J, Shin MK, Reinke C, Aggarwal NR, Jun JC,et al. Intermittent hypoxia inhibits clearance of triglyceride-rich lipoproteins and inactivates adipose lipoprotein lipase in a mouse model of sleep apnoea. Eur Heart J 2012;33:783-790.
Drager LF, Yao QL, Hernandez KL, Shin MK, Bevans-Fonti S, Gay J, et al. Chronic intermittent hypoxia induces atherosclerosis via activation of adipose angiopoietin-like 4.Am J Respir Crit Care Med 2013;188:240-248.
Manalo DJ, Rowan A, Lavoie T, Natarajan L, Kelly BD, Ye SQ, et al. Transcriptional regulation of vascular endothelial cell responses to hypoxia by HIF-1. Blood 2005;105:659-669.
Zhang H, Wong CC, Wei H, Gilkes DM, Korangath P, Chaturvedi P, et al. HIF-1-dependent expression of angiopoietin-like 4 and L1CAM mediates vascular metastasis of hypoxic breast cancer cells to the lungs.Oncogene 2012;31:1757-1770.
Semenza GL. Targeting HIF-1 for cancer therapy. Natu Rev Cancer 2003;3:721-732.
Semenza GL. Defining the role of hypoxia-inducible factor 1 in cancer biology and therapeutics. Oncogene 2010;29:625.
Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, et al. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 2001;292:464-468.
Masson N, Willam C, Maxwell PH, Pugh CW, Ratcliffe PJ. Independent function of two destruction domains in hypoxia-inducible factor-α chains activated by prolyl hydroxylation. EMBO J 2014;20:5197-5206.
Liu Y, Ma Z, Zhao C, Wang Y, Wu G, Xiao J, et al. HIF-1α and HIF-2α are critically involved in hypoxia-induced lipid accumulation in hepatocytes through reducing PGC-1α-mediated fatty acid β-oxidation. Toxicol Lett 2014;226:117-123.
Arai T, Tanaka M, Goda N. HIF-1-dependent lipin1 induction prevents excessive lipid accumulation in choline-deficient diet-induced fatty liver. Sci Rep 2018;8:14230.
Salceda S, Caro J. Hypoxia-inducible factor 1alpha (HIF-1alpha) protein is rapidly degraded by the ubiquitin-proteasome system under normoxic conditions. Its stabilization by hypoxia depends on redox-induced changes. J Biol Chem 1997;272:22642-22647.
Wang SH, Wang J, Li J. Efficacy assessment of treating patients with coronary heart disease angina of phlegm and stasis mutual obstruction syndrome by Danlou Tablet. Chin J Integr Tradit West Med (Chin) 2012;32:1051.
Wang L, Mao S, Qi JY, Ren Y, Guo XF, Chen KJ, et al.Effect of Danlou Tablet on peri-procedural myocardial injury among patients undergoing percutaneous coronary intervention for non-ST elevation acute coronary syndrome:a study protocol of a multicenter, randomized, controlled trial. Chin J Integr Med 2015;21:662.
Miao J, Zhou XB, Mao W, Chen J, Xu XM. Effects of Xuefu Zhuyu Granule and Danlou Tablet on anti-atherosclerosis rats and potential mechanisms. Chin J Integr Tradit West Med (Chin) 2016;36:80-84.
Qi JY, Wang L, Gu DS, Guo LH, Zhu W, Zhang MZ.Protect effects of Danlou Tablet against murine myocardial ischemia and reperfusion injury in vivo. Chin J Integr Med 2016;24:1-8.
Wu XQ, Dong J, Fu AZ, Wu HH, Meng W, Ning L, et al.Studies of HPLC fingerprint of Danlou Tablets. Tradit Chin Drug Res Clin Pharm (Chin) 2014;25:319-322.
Nagel S, Papadakis M, Chen R, Hoyte LC, Brooks KJ,Gallichan D, et al. Neuroprotection by dimethyloxalylglycine following permanent and transient focal cerebral ischemia in rats. J Cereb Blood Flow Metab 2011;31:132-143.
Song YR, You SJ, Lee YM, Chin HJ, Chae DW, Oh YK,et al. Activation of hypoxia-inducible factor attenuates renal injury in rat remnant kidney. Nephrol Dial Transplant 2010;25:77-85.
Jaakkola PM, Mole D, Tian YM, Wilson MI. Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. Science 2001;292:468-472.
Asikainen TM, Schneider BK, Waleh NS, Clyman RI,Ho WB, Flippin LA, et al. Activation of hypoxia-inducible factors in hyperoxia through prolyl 4-hydroxylase blockade in cells and explants of primate lung. Proceedings of the National Academy of Sciences of the United States.2005;102:10212-10217.
Jawień J, Nastalek P, Korbut R. Mouse models of experimental atherosclerosis. J Physiol Pharm 2004;55:503-517.
Commission of Chinese Pharmacopoeia. Chinese Pharmacopoeia. Beijing: China Medical Science and Technology Press; 2015:681-683.
Desai U, Lee EC, Chung K, Gao C, Gay J, Key B, et al. Lipid-lowering effects of anti-angiopoietin-like 4 antibody recapitulate the lipid phenotype found in angiopoietin-like 4 knockout mice.Proc Natl Acad Sci USA 2007;104:11766-11771.
Jun J, Reinke C, Bedja D, Berkowitz D, Bevans-Fonti S,Li J, et al. Effect of intermittent hypoxia on atherosclerosis in apolipoprotein E-deficient mice. Atherosclerosis 2010;209:381-386.
Savransky V, Nanayakkara A, Li J, Bevans S, Smith PL, Rodriguez A, et al. Chronic intermittent hypoxia induces atherosclerosis. Am J Respir Crit Care Med 2007;175:1290-1297.
Zhang M, Zhao H, Cai J, Li H. Chronic administration of mitochondrion-targeted peptide SS-31 prevents atherosclerotic development in ApoE knockout mice fed Western diet. PLoS One 2017;12:e0185688.
Savransky V, Jun J, Li J, Nanayakkara A, Fonti S, Moser AB,et al. Dyslipidemia and atherosclerosis induced by chronic intermittent hypoxia are attenuated by deficiency of stearoyl coenzyme a desaturase. Circul Res 2008;103:1173-1180.
Alshehri KA, Bashamakh LF, Alshamrani HM. Pattern and severity of sleep apnea in a Saudi sleep center: the impact of obesity. J Fam Comm Med 2019;26:127-132.
Chen H, Hu K, Zhu J, Xianyu Y, Cao X, Kang J, et al.Polymorphisms of the 5-hydroxytryptamine 2A/2C receptor genes and 5-hydroxytryptamine transporter gene in Chinese patients with OSAHS. Sleep Breath 2013;17:1241-1248.
Kushida CA, Littner MR, Hirshkowitz M, Morgenthaler TI,Alessi CA, Bailey D, et al. Practice parameters for the use of continuous and bilevel positive airway pressure devices to treat adult patients with sleep-related breathing disorders.Sleep 2006;29:375.
Song D, Fang G, Mao SZ, Ye X, Liu G, Gong Y, et al.Chronic intermittent hypoxia induces atherosclerosis by NF-kappaB-dependent mechanisms. Biochim Biophys Acta 2012;1822:1650-1659.
Poitz DM, Augstein A, Hesse K, Christoph M, Ibrahim K,Braun-Dullaeus RC, et al. Regulation of the HIF-system in human macrophages-differential regulation of HIF-alpha subunits under sustained hypoxia. Mol Immunol 2014;57:226-235.
Nanduri J, Vaddi DR, Khan SA, Wang N, Makarenko V,Semenza GL, et al. HIF-1alpha activation by intermittent hypoxia requires NADPH oxidase stimulation by xanthine oxidase. PLoS One 2015;10:e0119762.
Drager LF, Qiaoling Y, Hernandez KL, Shin MK, Bevans-Fronti S, Gay J, et al. Chronic intermittent hypoxia induces atherosclerosis via activation of adipose angiopoietin-like 4.Am J Respir Crit Care Med 2013;188:240-248.
Yu C, Qi D, Lian W, Li QZ, Li HJ, Fan HY. Effects of danshensu on platelet aggregation and thrombosis: in vivo arteriovenous shunt and venous thrombosis models in rats.PLoS One 2014;9:e110124.
Zhang X, Wu Q, Lu Y, Wan J, Dai H, Zhou X, et al.Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways. Free Rad Biol Med 2018;124:504-516.
Zhang H, Wang Y, Gao C, Gu Y, Huang J, Wang J,et al. Salvianolic acid A attenuates kidney injury and inflammation by inhibiting NF-κB and p38 MAPK signaling pathways in 5/6 nephrectomized rats. Acta Pharmacol Sin 2018;39:1855-1864.
Li ZM, Xu SW, Liu PQ. Salvia miltiorrhiza Burge (Danshen):a golden herbal medicine in cardiovascular therapeutics.Acta Pharmacol Sin 2018;39:802-824.
Gao S, Liu Z, Li H, Little PJ, Liu P, Xu S. Cardiovascular actions and therapeutic potential of tanshinone ⅡA.Atherosclerosis 2012;220:3-10.
Xu S, Liu P. Tanshinone Ⅱ-A: new perspectives for old remedies. Exp Opin Ther Patents 2013;23:149-153.
Yan H, Zou C. Network pharmacology of triterpenes and their glycosides in Fructus Trichosanthis. J Intern Pharm Res (Chin) 2018;4:288-294.
Yu X, Tang L, Wu H, Zhang X, Luo H, Guo R, et al.Trichosanthis Fructus: botany, traditional uses, phytochemistry and pharmacology. J Ethnopharm 2018;224:177-194.
Wang PL, Li XC, Bai RN, Zang MJ. Danlou Tablet in the treatment of coronary heart disease with phlegm and stasis mutual obstruction syndrome: a meta-analysis. Chin J Integr Med Cardio-/Cerebrovasc Dis (Chin) 2015;13:2033-2037.
Liu JX, Lin CR, Ren JX, Li L, Ren JG, Fu JH, et al.Experimental study on pathogenetic evolvement regularity of phlegm, toxin and blood-stasis syndromes in Chinese miniswine with phlegm-stasis cementation syndrome of coronary heart disease. China J Chin Mater Med (Chin)2013;38:4138.
Tang L, Liu D, Yi X, Xu T, Liu Y, Luo Y, et al. The protective effects of puerarin in cardiomyocytes from anoxia/reoxygenation injury are mediated by PKC. Cell Biochem Funct 2014;32:378-386.
Zhang X, Wu Q, Lu Y, Wang J, Dai H, Zhou X, et al.Cerebroprotection by salvianolic acid B after experimental subarachnoid hemorrhage occurs via Nrf2- and SIRT1-dependent pathways. Radic Biol Med 2018;124:504-516.
Yin MS, Zhang YC, Xu SH, Liu JJ, Sun XH, Liang C, et al.Puerarin prevents diabetic cardiomyopathy in vivo and in vitro by inhibition of inflammation. J Asian Nat Prod Res 2019;21:476-493.
Yang Z, Hong T, Liu YM, Han Q. Protection of Danlou Tablets on hyperlipidemia and vascular endothelial injury in rats. World J Integr Tradit West Med (Chin) 2010;5:491-494.
Young SG, Zechner R. Biochemistry and pathophysiology of intravascular and intracellular lipolysis. Genes Dev 2013;27:459-484.
Dijk W, Beigneux AP, Larsson M, Bensadoun A, Young SG, Kersten S. Angiopoietin-like 4 promotes intracellular degradation of lipoprotein lipase in adipocytes. J Lipid Res 2016;57:1670-1683.
Kroupa O, Vorrsjö E, Stienstra R, Mattijssen F, Nilsson SK,Sukonina V, et al. Linking nutritional regulation of Angptl4,Gpihbp1, and Lmf1 to lipoprotein lipase activity in rodent adipose tissue. BMC Physiol 2012;12:13.
Gusarova V, O'dushlaine C, Teslovich TM, Benotti PN,Mirshahi T, Gottesman O, et al. Genetic inactivation of ANGPTL4 improves glucose homeostasis and is associated with reduced risk of diabetes. Nature Commun 2018;9:2252.
0
浏览量
0
Downloads
2
CSCD
关联资源
相关文章
相关作者
相关机构