HDAC9基因对LPS诱导冠状动脉平滑肌细胞增殖与凋亡及炎症反应影响

EFFECT OF THE HDAC9 GENE ON LIPOPOLYSACCHARIDE-INDUCED PROLIFERATION, APOPTOSIS, AND INFLAMMATORY RESPONSE OF CORONARY ARTERY SMOOTH MUSCLE CELLS

目的探讨蛋白去乙酰化酶9(HDAC9)基因对脂多糖(LPS)诱导的人冠状动脉平滑肌细胞(HCASMC)增殖、凋亡及炎症反应的影响。方法采用LPS诱导HCASMC,实验分为空白对照组、模型组、阴性对照组和转染组。采用实时荧光定量PCR(qPCR)和Western blot方法检测HDAC9 mRNA和蛋白表达;MTT法和流式细胞术分别检测细胞增殖和凋亡情况;ELISA检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-8(IL-8)含量。结果与空白对照组相比,模型组细胞中HDAC9 mRNA和蛋白表达水平升高(F=91.145、185.108,q=19.403、25.646,P<0.05)、细胞活力升高(F=77.940,q=17.819,P<0.05)、凋亡率降低(F=98.321,q=15.563,P<0.05)、TNF-α与IL-1β及IL-8的含量增多(F=91.145~325.808,q=10.813~15.180,P<0.05)。敲低HDAC9能够抑制细胞活力和炎症反应,并促进细胞凋亡。结论敲低HDAC9可抑制LPS诱导的HCASMC增殖及炎症反应,并促进细胞凋亡。

Objective To investigate the effect of the protein deacetylase 9(HDAC9)gene on lipopolysaccharide(LPS)-induced proliferation,apoptosis,and inflammatory response of human coronary artery smooth muscle cells(HCASMCs).Me-thods HCASMCs were induced by LPS,and the cells were divided into blank control group,model group,negative control group,and transfection group.Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of HDAC9;MTT assay and flow cytometry were used to measure cell proliferation and apoptosis;ELISA was used to mea-sure the levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-8(IL-8).Results Compared with the blank control group,the model group had significant increases in the mRNA and protein expression levels of HDAC9 in cells(F=91.145-185.108,q=19.403-25.646,P<0.05),a significant increase in cell viability(F=77.940,q=17.819,P<0.05),a significant reduction in cell apoptosis rate(F=98.321,q=15.563,P<0.05),and significant increases in the levels of TNF-α,IL-1β,and IL-8(F=91.145-325.808,q=10.813-15.180,P<0.05).HDAC9 knockdown inhibited cell viability and inflammatory response and promoted cell apoptosis.Conclusion HDAC9 knockdown can inhibit LPS-induced proliferation and inflammation of HCASMCs and promote cell apoptosis.