天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (10): 1045-1049.doi: 10.11958/20190295

• 实验研究 • 上一篇    下一篇

CDK5介导的PPARγ磷酸化在动脉粥样硬化泡沫细胞形成过程中的作用

沈娜 1，2，贺晶 3，邸研博 2，刘勇 3，田凤石 3，刘运德 1△   

  1. 1天津医科大学医学检验学院（邮编300203）；2天津市第四中心医院中心实验室，3心血管内科
  • 收稿日期:2019-01-29 修回日期:2019-03-28 出版日期:2019-10-15 发布日期:2019-11-11
  • 通讯作者: 田凤石 E-mail:daixiezonghezheng@163.com
  • 基金资助:
    中国中青年临床研究基金-VG基金;天津市卫生行业重点攻关项目;天津市慢性病防治科技重大专项

The role of CDK5-mediated PPARγ phosphorylation in the formation of foam cells in atherosclerosis

SHEN Na1,2，HE Jing3，DI Yan-bo2，LIU Yong3，TIAN Feng-shi3，LIU Yun-de1△   

  1. 1 School of Medical Laboratory, Tianjin Medical University, Tianjin 300203, China; 2 Central Laboratory, 3 Department of Cardiology, Tianjin 4th Center Hospital
  • Received:2019-01-29 Revised:2019-03-28 Published:2019-10-15 Online:2019-11-11
  • Supported by:
     

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摘要： 摘要：目的 探讨细胞周期素依赖蛋白激酶5（CDK5）介导的过氧化物酶体增殖物激活受体γ（PPARγ）磷酸化在 动脉粥样硬化中的作用。方法 常规培养小鼠Raw264.7巨噬细胞，实验设对照组（C组）、氧化低密度脂蛋白（oxLDL）组（O组，50 mg/L ox-LDL）、Roscovitine+ox-LDL组（R组，15 μmol/L Roscovitine+50 mg/L ox-LDL）。待细胞融合 至70%左右，R组加入15 μmol/L Roscovitine预处理30 min，之后O组和R组分别加入50 mg/L ox-LDL继续培养24 h， 使其转化为泡沫细胞；C组不作处理。利用Western blot检测各组pPPARγ、tPPARγ、p35和CDK5蛋白表达的变化，油 红O染色和异丙醇萃取实验分析各组细胞内脂质聚积情况，酶法测定细胞内胆固醇含量，反转录PCR（RT-PCR）检 测各组ox-LDL摄取相关基因CD36、SR-A1和胆固醇外流相关基因ABCA1、ABCG1的mRNA表达水平。结果 oxLDL诱导后，O组pPPARγ/tPPARγ比值、p35/CDK5比值、细胞内脂质聚积、总胆固醇含量、游离胆固醇含量、胆固醇 酯/总胆固醇比值均较 C 组明显升高（P＜0.05）；CDK5 抑制剂干预后，R 组上述指标均较 O 组降低（P＜0.05）。RTPCR结果显示，ox-LDL诱导后，O组ox-LDL摄取相关基因CD36、SR-A1的mRNA表达水平升高，而胆固醇外流相关 基因ABCA1、ABCG1的mRNA表达水平降低（P＜0.05）；CDK5抑制剂干预后，上述指标变化与O组呈相反趋势（P＜ 0.05）。结论 CDK5/pPPARγ途径参与动脉粥样硬化泡沫细胞的形成。

关键词: 动脉粥样硬化, 泡沫细胞, 细胞周期蛋白依赖激酶 5, 过氧化物酶体增殖物激活受体, CD36, SR-A1, ABCA1, ABCG1

Abstract: Abstract: Objective To investigate the role of cyclin-dependent kinases 5 (CDK5) - mediated PPARγ phosphorylation in atherosclerosis. Methods Raw264.7 macrophages were induced by ox-LDL to differentiate into foam cells, and then CDK5 inhibitor was used as the intervent. The experimental cells were divided into control group (C group), ox-LDL group (O group, 50 mg/L ox-LDL) and roscovitine group (50 mg/L ox-LDL+15 μmol/L Roscovitine). Expressions of pPPARγ, tPPARγ, p35 and CDK5 protein were detected by Western blot assay. Lipid accumulation was analyzed by oil red O staining and isopropyl alcohol extraction experiments. Levels of cholesterol content in macrophages were measured by enzymatic method. Expression of ox-LDL uptake related genes CD36, SR-A1 and cholesterol efflux related genes ABCA1 and ABCG1 were detected by RT-PCR. Results After ox-LDL induction, the ratios of pPPARγ/tPPARγ and p35/CDK5 significantly increased, and lipid accumulation, total cholesterol content, free cholesterol content and the ratio of cholesterol ester / total cholesterol were up-regulated significantly in O group compared with those of C group (P＜0.05). RT-PCR showed that expression levels of ox-LDL uptake-related genes CD36 and SR-A1 mRNA increased, while the expression levels of cholesterol efflux-related genes ABCA1 and ABCG1 decreased in O group (P＜0.05). After intervention with CDK5 inhibitor, the above indicators showed the opposite trend compared with those of O group (P＜0.05). Conclusion CDK5/ pPPARγ pathway is involved in the formation of foam cells in atherosclerosis.

Key words: atherosclerosis, foam cells, cyclin-dependent kinase 5, peroxisome proliferator-activated receptors, CD36, SR-A1, ABCA1, ABCG1

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