天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (11): 1025-1030.doi: 10.11958/20200939

• 细胞与分子生物学 •    下一篇

IL-6、GATA6在大鼠肺动脉平滑肌细胞中的表达及对 细胞增殖的影响

罗杰1，刘维佳2△   

  1. 1贵州大学医学院（邮编550025）；2贵州省人民医院
  • 收稿日期:2020-04-10 修回日期:2020-08-24 出版日期:2020-11-15 发布日期:2020-11-15
  • 基金资助:
    贵州省科技计划项目（黔科合基础[2018]1408）

Expressions of IL-6 and GATA6 in rat pulmonary artery smooth muscle cells and their effects on cell proliferation

LUO Jie1， LIU Wei-jia2△   

  1. 1 School of Medicine, Guizhou University, Guiyang 550025, China; 2 Guizhou Provincial People's Hospital
  • Received:2020-04-10 Revised:2020-08-24 Published:2020-11-15 Online:2020-11-15

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摘要： 目的　探讨白细胞介素（IL）-6、转录因子GATA6在血小板源性生长因子-BB（PDGF-BB）诱导大鼠肺动脉平滑肌细胞（PASMCs）增殖中的表达及作用。方法　从8周龄SD大鼠中分离肺动脉，采用Ⅰ型胶原酶消化法原代培养PASMCs；采用倒置显微镜观察和免疫荧光染色鉴定细胞；采用CCK-8检测不同剂量（0、15、30、45、60、75 μg/L）PDGF-BB分别作用12、24、48 h对PASMCs增殖能力的影响；采用流式细胞术检测30 μg/L PDGF-BB诱导细胞增殖48 h的细胞周期；应用qPCR和Western blot技术检测30 μg/L PDGF-BB诱导细胞增殖12、24、48 h后IL-6和GATA6 mRNA及蛋白的表达水平。结果　15~60 μg/L的PDGF-BB诱导24 h、48 h后PASMCs增殖显著，其中以30 μg/L诱导48 h效果最佳（P＜0.05）；流式周期结果表明，PDGF-BB组G1期的百分比较对照组降低，而S期和G2期百分比增高（P＜0.01）。PDGF-BB能够诱导细胞增加IL-6 mRNA和蛋白的分泌并降低GATA6 mRNA和蛋白表达水平。相关性分析提示PASMCs的增殖与IL-6 mRNA表达呈正相关，与GATA6 mRNA表达呈负相关（r分别为0.538和-0.647，均P＜0.05）；IL-6与GATA6的表达呈负相关（r=-0.705，P＜0.01）。结论　PDGF-BB诱导体外培养的PASMCs增殖；IL-6、GATA6参与了PDGF-BB诱导PASMCs增殖的发生和发展；PASMC增殖与IL-6 mRNA表达上调和GATA6 mRNA表达下调有关。

关键词: 血小板源性生长因子；白细胞介素6；GATA6转录因子；肌细胞, 平滑肌；细胞增殖

Abstract: Objective　To explore the expressions and roles of interleukin (IL)-6 and transcription factor GATA6 in the proliferation of rat pulmonary artery smooth muscle cells (PASMCs) induced by platelet-derived growth factor-BB (PDGF-BB). Methods　Pulmonary arteries were isolated from 8-week-old SD rats, and PASMCs were cultured by type Ⅰ collagenase digestion method. The cells were observed by inverted microscope and identified by immunofluorescence staining. The effects of different concentrations of PDGF-BB (0, 15, 30, 45, 60 and 75 μg/L) on the proliferation of PASMCs at 12 h, 24 h and 48 h were detected by CCK-8. Flow cytometry was used to detect the cell cycle of 30 μg/L PDGF-BB induced cell proliferation for 48 h. The qPCR and Western blot assay were used to detect the expression levels of IL-6 mRNA and GATA6 mRNA and protein after 30 μg/L PDGF-BB induced cell proliferation for 12 h, 24 h and 48 h. Results　The proliferation model of PASMCs could be effectively established by PDGF-BB. The proliferation of PASMCs was significantly increased by PDGF-BB at 15-60 μg/L for 24 h and 48 h, and the best effect was induced by 30 μg/L for 48 h (P＜0.05). Flow cytometry results showed that the percentage of G1 phase was significantly lower in PDGF-BB group than that of control group, while the percentages of S phase and G2 phase were significantly increased (P＜0.01). PDGF-BB can induce cells to increase IL-6 mRNA and protein secretion and reduce GATA6 mRNA and protein expression levels. Correlation analysis showed that the proliferation of PASMCs was positively correlated with the expressions of IL-6 mRNA and negatively correlated with the expression of GATA6 mRNA (r=0.538, -0.647 respectively, P＜0.05). The expression of IL-6 mRNA was negatively correlated with the expression of GATA6 mRNA (r=-0.705, P＜0.01). Conclusion　PDGF-BB induces proliferation of PASMCs cultured in vitro. IL-6 and GATA6 are involved in the occurrence and development of PDGF-induced PASMCs proliferation. PASMC proliferation is associated with up-regulation of IL-6 mRNA and down-regulation of GATA6 mRNA.

Key words: platelet-derived growth factor, interleukin-6, GATA6 transcription factor, myocytes, smooth muscle, cell proliferation