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目的研究小檗碱(Ber)对内脏脂肪素(visfatin)诱导人脐静脉内皮细胞(HUVEC)损伤的保护作用及与p38丝裂原活化蛋白激酶(p38 MAPK)通路的关系。方法以visfatin不同质量浓度(0,50,100,200μg·L-1)及以100μg·L-1质量浓度的不同时间(0,6,12,24,48 h)作用HUVEC,MTT法检测细胞增殖;流式细胞术检测细胞凋亡;ELISA法检测白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)含量;Western blot法检测HUVEC中p-p38 MAPK、Bax、Bcl-2蛋白表达;并以50μmol·L-1 Ber及20μmol·L-1 SB203580(P38分裂原激活蛋白激酶通路抑制剂)检测Ber的干预作用。结果与对照组比较,100μg·L-1 visfatin可显著抑制HUVEC增殖,诱导HUVEC分泌IL-6及TNF-α,上调HUVEC内p-p38 MAPK、Bax蛋白表达及降低Bcl-2蛋白表达以促进细胞凋亡(P<0.01);与visfatin组比较,Ber可显著减轻visfatin对HUVEC增殖的抑制、下调HUVEC内p-p38 MAPK、Bax蛋白表达及上调Bcl-2蛋白表达以抑制HUVEC凋亡、减少visfatin诱导HUVEC分泌IL-6及TNF-α(P<0.01)。结论 Ber可减轻visfatin诱导的HUVEC损伤,其机制与抑制p38 MAPK通路有关。
Objective To investigate the protective effect of berberine on the visfatin-induced injury of human umbilical vein endothelial cells (HUVECs) and its relationship with the p38 mitogen activated protein kinase (p38 MAPK) pathway. Methods HUVECs were treated with different concentrations of visfatin (0, 50, 100, 200μg · L-1) and different concentrations of 100μg · L-1 for 0, 6, 12, 24 and 48 h. MTT assay was used to detect cell proliferation. The levels of IL-6 and TNF-α were detected by ELISA. The expressions of p-p38 MAPK, Bax and Bcl-2 in HUVEC were detected by Western blot. The protein expression of Ber was detected by Western blotting. The effects of Ber on the expression of Ber34 in 50μmol·L-1 Ber and 20μmol·L-1 SB203580 (P38 mitogen-activated protein kinase pathway inhibitor) were detected. Results Compared with the control group, 100μg · L-1 visfatin significantly inhibited the proliferation of HUVECs and induced the secretion of IL-6 and TNF-α by HUVECs and up-regulated the expressions of p-p38 MAPK and Bax protein and decreased the expression of Bcl-2 protein in HUVEC (P <0.01). Compared with visfatin group, Ber could significantly reduce the inhibition of visfatin on the proliferation of HUVEC, down-regulate the expression of p-p38 MAPK, Bax protein and up-regulate the expression of Bcl-2 protein in HUVEC to inhibit the apoptosis of HUVEC and decrease visfatin Induction of HUVEC secretion of IL-6 and TNF-α (P <0.01). Conclusion Ber can reduce the visfatin-induced HUVEC injury and its mechanism is related to the inhibition of the p38 MAPK pathway.