PPAR－γ激动剂对炎症状态下人系膜细胞LOX－1表达的影响

doi:10.3877/cma.j.issn.2095-3216.2019.06.007

中华肾病研究电子杂志 ›› 2019, Vol. 08 ›› Issue (06) : 263 -268. doi: 10.3877/cma.j.issn.2095-3216.2019.06.007

所属专题：文献；

论著

PPAR－γ激动剂对炎症状态下人系膜细胞LOX－1表达的影响

刘华¹, 李银平¹, 吴雷云¹, 林娜¹, 付文静¹, 李文¹, 董星彤¹, 贾林沛¹, 贾强¹, 邓英辉¹^,^†(

)

^1. 100053　北京，首都医科大学宣武医院肾内科

收稿日期:2019-03-11 出版日期:2019-12-28
通信作者: 邓英辉

Effect of PPAR-γ agonist on the expression of LOX-1 in human mesangial cells under inflammatory conditions

Hua Liu¹, Yinping Li¹, Leiyun Wu¹, Na Lin¹, Wenjing Fu¹, Wen Li¹, Xingtong Dong¹, Linpei Jia¹, Qiang Jia¹, Yinghui Deng¹^,^†()

^1. Department of Nephrology, Xuanwu Hospital, Capital Medical University, Beijing 100053, China

Received:2019-03-11 Published:2019-12-28
Corresponding author: Yinghui Deng
About author:
Corresponding author: Deng Yinghui, Email: yinghuideng20046@sina.com

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目的

研究过氧化物酶体增殖体激活受体γ(PPAR－γ)激动剂15－脱氧前列腺素J₂(15d－PGJ₂)对炎症因子白细胞介素1β(IL－1β)作用下人肾小球系膜细胞(HMC)氧化低密度脂蛋白(Ox－LDL)植物血凝素样受体1(LOX－1)表达的影响。

方法

激光共聚焦显微镜观察IL－1β对HMC脂质摄取的影响；实时定量PCR和Western印迹方法检测不同剂量的PPAR－γ激动剂15d－PGJ₂对IL－1β诱导的LOX－1表达的影响。

结果

激光共聚焦显微镜检查发现5 ng/ml的IL－1β促进HMC摄取荧光标记的Ox－LDL；IL－1β能够促进HMC的LOX－1 mRNA和蛋白表达，15d－PGJ₂呈剂量依赖性抑制IL－1β诱导的LOX－1表达。与单纯IL－1β(5 ng/ml)刺激组相比，15d－PGJ₂浓度2.5、5、10 μmol/L处理组细胞LOX－1 mRNA表达分别下调为73.13%、66.54%、35.23%；15d－PGJ₂浓度5、10 μmol/L处理组细胞LOX－1蛋白表达分别下调为82.39%、63.17%。

结论

炎症因子IL－1β促进HMC摄取Ox－LDL；PPAR－γ激动剂15d－PGJ₂剂量依赖性抑制IL－1β诱导的LOX－1 mRNA和蛋白表达，对炎症因子导致的系膜细胞脂质失衡可能具有保护作用。

关键词: 过氧化物酶体增殖体激活受体γ激动剂, 白细胞介素1β, 氧化低密度脂蛋白植物血凝素样受体1

Objective

To investigate the effect of peroxisome proliferator-activated receptor γ (PPAR-γ) agonist 15-deoxyprostaglandin J₂ (15d-PGJ₂) on the expression of lectin-like oxidized low-density lipoprotein (Ox-LDL) receptor 1 (LOX-1) in human glomerular mesangial cells (HMC) treated with interleukin 1β (IL-1β).

Methods

Laser confocal microscopy was used to observe the effect of IL-1β on lipid uptake by HMC. Real-time quantitative PCR and Western blotting were applied to study the effects of different doses of PPAR-γ agonist 15d-PGJ₂ on IL-1β-induced LOX-1 expression.

Results

Laser confocal microscopy revealed that IL-1β (5 ng/ml) promoted HMC uptake of Ox-LDL. IL-1β promoted the mRNA and protein expression of LOX-1 in HMC, while 15d-PGJ₂ dose-dependently inhibited IL-1β-induced LOX-1 expression. Compared with the IL-1β (5ng/ml) group, the treatment group showed that 15d-PGJ₂ in concentrations of 2.5 μmol/L, 5 μmol/L, and 10 μmol/L down-regulated the mRNA expression of LOX-1 to 73.13%, 66.54%, and 35.23%, respectively, while 15d-PGJ₂ in concentrations of 5 μmol/L and 10 μmol/L down-regulated the protein expression of LOX-1 to 82.39% and 63.17%, respectively.

Conclusion

Inflammatory factor IL-1β promoted the uptake of Ox-LDL and expression of LOX-1 in HMC. PPAR-γ agonist 15d-PGJ₂ dose-dependently inhibited the mRNA and protein expression of LOX-1 induced by IL-1β, which may have a protective role in the lipid imbalance of HMC caused by inflammatory factors.

Key words: PPAR-γ agonist, Interleukin-1β, Lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1)

图1 IL－1β对HMC摄取Dil－Ox－LDL的影响(400×)

图2 15d－PGJ₂　的细胞毒性试验

表1 15d－PGJ₂的细胞毒性试验

15d－PGJ₂浓度(μmol/L)	细胞死亡比率(%)(n＝6)
15d－PGJ₂浓度(μmol/L)	台盘兰试验	LDH释放试验
0	0.32±0.13	0.48±0.15
2.5	1.26±0.15	1.75±0.13
5	2.85±0.27	3.26±0.21
10	3.86±0.21	4.17±0.18
15	16.78±0.13	19.83±0.16
20	31.25±0.26	41.08±0.23

表1 15d－PGJ₂的细胞毒性试验

15d－PGJ₂浓度(μmol/L)	细胞死亡比率(%)(n＝6)
15d－PGJ₂浓度(μmol/L)	台盘兰试验	LDH释放试验
0	0.32±0.13	0.48±0.15
2.5	1.26±0.15	1.75±0.13
5	2.85±0.27	3.26±0.21
10	3.86±0.21	4.17±0.18
15	16.78±0.13	19.83±0.16
20	31.25±0.26	41.08±0.23

图3 15d－PGJ₂　对IL－1β诱导的LOX－1mRNA表达的影响

图4 15d－PGJ₂　对IL－1β诱导的LOX－1蛋白表达的影响

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