补肾降脂方调控自噬与胆固醇流出干预ApoE-/-小鼠动脉粥样硬化的机制研究

摘要
图/表
参考文献(20)
相关文章 (15)

全文: PDF (1006 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的探讨补肾降脂方调控自噬与胆固醇流出干预动脉粥样硬化（AS）的作用机制。方法高脂饮食诱导100只ApoE-/-小鼠复制AS模型，采用随机数字表法分为模型组、补肾组、他汀组、3-甲基腺嘌呤（3-MA）组，每组25只；另选25只C57BL/6J小鼠为正常对照。苏木精-伊红（HE）、油红O及FILIPIN染色检测主动脉窦病理形态、脂质蓄积及游离胆固醇（FC）染色情况；电镜观察自噬小体、脂滴；比色法检测血清总胆固醇（TC）、FC含量；Western blot检测主动脉肝X受体α（LXRα）、ATP结合膜盒转运蛋白A1（ABCA1）、微管相关蛋白1轻链3（LC3）及p62蛋白表达。结果模型组主动脉窦AS斑块、脂质蓄积及FC阳性染色面积多于正常组，差异有高度统计学意义（P ＜ 0.01）；正常组主动脉内皮细胞结构完整，可见自噬小体；模型组主动脉内皮细胞肿胀，可见脂滴，未见自噬小体；模型组TC、FC、胆固醇酯（CE）含量高于正常组，差异有高度统计学意义（P ＜ 0.01）；模型组主动脉LXRα、ABCA1、LC3Ⅱ/Ⅰ表达低于正常组，p62表达高于正常组，差异有高度统计学意义（P ＜ 0.01）；补肾组主动脉窦AS斑块、脂质蓄积及FC阳性染色面积少于模型组，差异有高度统计学意义（P ＜ 0.01）；补肾组主动脉内皮细胞可见自噬小体；补肾组TC、FC、CE含量低于模型组，差异有统计学意义（P ＜ 0.05或P ＜ 0.01）；补肾组LXRα、ABCA1、LC3Ⅱ/Ⅰ表达高于模型组，p62表达低于模型组，差异有统计学意义（P ＜ 0.05或P ＜ 0.01）。结论补肾降脂方可能通过调控自噬与胆固醇流出进而有效干预治疗AS。

	服务

	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	闫黎申定珠

关键词 ：补肾降脂方, 动脉粥样硬化, 自噬, 胆固醇流出

Abstract：Objective To explore the mechanism of Bushen Jiangzhi Formula in regulating autophagy and cholesterol efflux in the intervention of atherosclerosis (AS). Methods One hundred ApoE-/- mice induced by high fat diet were divided into model group, kidney tonifying group, statin group and 3-methyladenine (3-MA) group by using random number table method, with 25 mice in each group. Twenty-five C57BL/6J mice were normal controls. Hematoxylin-eosin (HE), oil red O and FILIPIN staining were used to detect the pathological morphology, lipid accumulation and free cholesterol (FC) staining of the aortic sinus. Autophagosomes and lipid droplets were observed by electron microscopy. The content of serum total cholesterol (TC) and FC were detected by colorimetry. The expression of liver X receptor alpha (LXRα), ATP-binding membrane cassette transport protein A1 (ABCA1), microtubule-associated protein 1 light chain 3 (LC3) and p62 protein were detected by Western blot. Results The model group had more aortic sinus AS plaque, lipid accumulation and FC positive staining area than the normal group, and the differences were highly statistically significant (P ＜ 0.01). The normal group had intact aortic endothelial cell structure and autophagosome. The endothelial cells of aorta in model group were swollen with lipid droplets, but no autophagosomes were observed. The contents of TC, FC, and cholesterol ester (CE) in model group were higher than those in normal group, and the differences were highly statistically significant (P ＜ 0.01). The expression of LXRα, ABCA1, and LC3 Ⅱ/Ⅰ in aorta of model group was lower than that of normal group, and the expression of p62 was higher than that of normal group, the differences were highly statistically significant (P ＜ 0.01). AS plaque, lipid accumulation, and FC positive staining area of aortic sinus in the kidney tonifying group were less than those in the model group, and the differences were highly statistically significant (P ＜ 0.01). In the kidney tonifying group, autophagosomes were found in the endothelial cells of aorta. The contents of TC, FC, and CE in kidney tonifying group were lower than those in model group, and the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01). The expression of LXRα, ABCA1, LC3Ⅱ/Ⅰ in kidney tonifying group was higher than that in model group, while the expression of p62 was lower than that in model group, the differences were statistically significant (P ＜ 0.05 or P ＜ 0.01). Conclusion Bushen Jiangzhi Formula may effectively intervene in the treatment of AS by regulating autophagy and cholesterol efflux.

Key words： Bushen Jiangzhi Formula Atherosclerosis Autophagy Cholesterol efflux

基金资助:国家自然科学基金面上项目（82074506、818733 48）；
全国中医药创新骨干人才培训项目（国中医药人教函〔2019〕128号）；
上海中医药大学“研究生创新培养专项”（Y201934）。

通讯作者: 申定珠（1975-），女，医学博士，主任医师，硕士生导师；研究方向：中医药防治老年病临床与基础研究。

作者简介: 闫黎（1993-），女，上海中医药大学龙华临床医学院2018级中医内科学专业在读硕士研究生；研究方向：中医药防治老年病临床与基础研究。

引用本文:

闫黎申定珠. 补肾降脂方调控自噬与胆固醇流出干预ApoE-/-小鼠动脉粥样硬化的机制研究[J]. 中国医药导报, 2021, 18(33): 7-11.
YAN Li SHEN Dingzhu. Study on the mechanism of Bushen Jiangzhi Formula regulating autophagy and cholesterol efflux on the intervention of atherosclerosis in ApoE-/- mice#br#. 中国医药导报, 2021, 18(33): 7-11.

链接本文:

https://www.yiyaodaobao.com.cn/CN/ 或 https://www.yiyaodaobao.com.cn/CN/Y2021/V18/I33/7

[1] Castano D，Rattanasopa C，Monteiro-Cardoso VF，et al. Lipid efflux mechanisms， relation to disease and potential therapeutic aspects [J]. Adv Drug Deliv Rev，2020，159：54-93.
[2] Guo S，Li L，Yin H. Cholesterol Homeostasis and Liver X Receptor（LXR）in Atherosclerosis [J]. Cardiovasc Hematol Disord Drug Targets，2018，18（1）：27-33.
[3] Okamoto Y，Tomioka M，Ogasawara F，et al. C-terminal of ABCA1 separately regulates cholesterol floppase activity and cholesterol efflux activity [J]. Biosci Biotechnol Biochem，2020，84（4）：764-773.
[4] Wang D，Hiebl V，Schachner D，et al. Soraphen A enhances macrophage cholesterol efflux via indirect LXR activation and ABCA1 upregulation [J]. Biochem Pharmacol，2020， 177：114022.
[5] Khawar MB，Gao H，Li W. Autophagy and Lipid Metaboli-sm [J]. Adv Exp Med Biol，2019，1206：359-374.
[6] 申定珠，陈川，迟惠英，等.从血管老化角度探讨动脉粥样硬化中医防治策略[J].中国中西医结合杂志，2012，32（2）：266-268.
[7] 贾庆玲，申定珠.补肾降脂方通过PPARγ-LXRα-ABCA1通路干预ApoE-/-小鼠动脉粥样硬化的效应机制[J].上海中医药杂志，2020，54（4）：85-91.
[8] Cao H，Jia QL，Shen DZ，et al. Bushen Jiangzhi formula reduces atherosclerosis in apoE-/- mice through autophagy [J]. J Tradit Chin Med，2020，40（4）：593-601.
[9] Wang B，Tontonoz P. Liver X receptors in lipid signalling and membrane homeostasis [J]. Nat Rev Endocrinol，2018， 14（8）： 452-463.
[10] Ouimet M，Barrett TJ，Fisher EA. HDL and Reverse Cholesterol Transport [J]. Circ Res，2019，124（10）：1505-1518.
[11] Wang N，Westerterp M. ABC Transporters，Cholesterol Efflux，and Implications for Cardiovascular Diseases [J]. Adv Exp Med Biol，2020，1276：67-83.
[12] Breslow JL. Mouse Models of Atherosclerosis [J]. Science，1996，272（5262）：685-688.
[13] Mialet-Perez J，Vindis C. Autophagy in health and disease：focus on the cardiovascular system [J]. Essays Biochem，2017，61（6）：721-732.
[14] Nadal M，Gold SE. Assessment of autophagosome formation by transmission electron microscopy [J]. Methods Mol Biol，2012，835：481-489.
[15] Wang XF，Qi HB，Wang Q，et al. FGFR3/fibroblast growth factor receptor 3 inhibits autophagy through decreasing the ATG12-ATG5 conjugate，leading to the delay of cartilage development in achondroplasia [J]. Autophagy，2015，11（11）：1998-2013.
[16] Jiang P，Mizushima N. LC3-and p62-based biochemical methods for the analysis of autophagy progression in mammalian cells [J]. Methods，2015，75：13-18.
[17] Ouimet M，Franklin V，Mak E，et al. Autophagy regulates cholesterol efflux from macrophage foam cells via lysosomal acid lipase [J]. Cell Metab，2011，13（6）：655-667.
[18] Shi YN，Jiang S，Zhao TJ，et al. Celastrol suppresses lipid accumulation through LXRα/ABCA1 signaling pathway and autophagy in vascular smooth muscle cells [J]. Biochem Biophys Res Commun，2020，532（3）：466-474.
[19] Lin XL，Guo DM，Liu MH，et al. FGF21 induces autophagy-mediated cholesterol efflux to inhibit atherogenesis via RACK1 up-regulation [J]. J Cell Mol Med，2020，24（9）：4992-5006.
[20] 闫黎，贾庆玲，申定珠.从脂噬角度探讨补肾中药复方防治动脉粥样硬化的新策略[J].中国医药导报，2019， 16（30）：159-162.