Wan Qiang, Yang Yu-Ping, Liu Zhong-Yong
Department of Medical Cardiology, the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
Department of Respiratory Medicine, the Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang 330006, China.
Zhongguo Zhong Yao Za Zhi. 2016 Jun;41(12):2309-2314. doi: 10.4268/cjcmm20161223.
To investigate the effect and the mechanism of puerarin in attenuating PM2.5-induced human umbilical vein endothelial cells (EA.hy926) injury, the samples of fine particulate matter (PM2.5) were collected and made into suspension. Different concentrations of PM2.5 (0,20, 200, 400 mg•L⁻¹) were used to contaminate EA.hy926 cells for 24 h. The cells survival rate was detected by MTT assay; cells apoptosis of EA.hy926cells was detected by flow cytometry; the protein levels of p-ERK1/2, Bax and Bcl-2 were detected by Western blot; the contents of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), malonaldehyde (MDA), and the activities of superoxide dismutase (SOD) and lactic dehydrogenase (LDH) were measured by ELISA. Puerarin at different concentrations (10, 50, 100 μmol•L⁻¹) or a specific inhibitor of ERK1/2 pathway PD98059 (20 μmol•L-1) was added into the EA.hy926 cells to observe the intervention effect and mechanism of puerarin. Compared with the control group, PM2.5 reduced the cells survival rate, up-regulatedp-ERK1/2 protein level and Bax/Bcl-2 ratio in a dose dependent manner to promote apoptosis; increased the contents of TNF-α, IL-6 and MDA, the activity of LDH, but decreased SOD activity in the EA.hy926 cells (P<0.05). Compared with PM2.5 group, puerarin increased the cells survival rate, down-regulated p-ERK1/2 protein level and Bax/Bcl-2 ratio in a dose dependent manner to inhibit the apoptosis; decreased the contents of TNF-α, IL-6 and MDA, the activity of LDH, but increased SOD activity in the EA.hy926 cells (P<0.05). The results indicated that puerarin could attenuate PM2.5-induced EA.hy926 cells injury via the inhibition of ERK1/2 pathway.
为探讨葛根素减轻细颗粒物（PM2.5）诱导的人脐静脉内皮细胞（EA.hy926）损伤的作用及机制，收集细颗粒物（PM2.5）样本并制成悬液。采用不同浓度的PM2.5（0、20、200、400mg•L⁻¹）污染EA.hy926细胞24小时。通过MTT法检测细胞存活率；采用流式细胞术检测EA.hy926细胞凋亡情况；通过蛋白质免疫印迹法检测p-ERK1/2、Bax和Bcl-2蛋白水平；采用酶联免疫吸附测定法检测肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、丙二醛（MDA）含量以及超氧化物歧化酶（SOD）和乳酸脱氢酶（LDH）活性。将不同浓度（10、50、100μmol•L⁻¹）的葛根素或ERK1/2通路特异性抑制剂PD98059（20μmol•L-1）加入EA.hy926细胞中，观察葛根素的干预作用及机制。与对照组相比，PM2.5呈剂量依赖性降低细胞存活率，上调p-ERK1/2蛋白水平及Bax/Bcl-2比值以促进细胞凋亡；增加EA.hy926细胞中TNF-α、IL-6和MDA含量以及LDH活性，但降低SOD活性（P<0.05）。与PM2.5组相比，葛根素呈剂量依赖性提高细胞存活率，下调p-ERK1/2蛋白水平及Bax/Bcl-2比值以抑制细胞凋亡；降低EA.hy926细胞中TNF-α、IL-6和MDA含量以及LDH活性，但增加SOD活性（P<0.05）。结果表明，葛根素可通过抑制ERK1/2通路减轻PM2.5诱导的EA.hy926细胞损伤。
