目的 研究c-Jun氨基端激酶(JNK)信号转导通路对内脏脂肪素(内脂素)诱导人脐静脉内皮细胞(HUVEC)分泌白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的调控作用及小檗碱的干预作用。方法 体外培养HUVEC, 用内脂素不同质量浓度(0、50、100、200 μg/L)及不同时间(0、6、12、24、48 h)作用HUVEC, MTT法检测HUVEC增殖;流式细胞仪检测HUVEC凋亡;ELISA法检测HUVEC上清液中IL-6及TNF-α的量;Western blotting法检测HUVEC中p-JNK、Bax、Bcl-2蛋白表达;并用小檗碱(50 μmol/L)及JNK通路特异性抑制剂SP600125(10 μmol/L)干预HUVEC, 检测小檗碱的干预作用。结果 与对照组相比, 100 μg/L内脂素可显著抑制HUVEC增殖, 诱导HUVEC凋亡, 诱导HUVEC分泌IL-6及TNF-α 的量显著增高, 并可显著增加HUVEC内p-JNK及Bax蛋白表达, 减少HUVEC内Bcl-2蛋白表达;与内脂素组相比, 小檗碱及SP600125均可显著减轻内脂素对HUVEC增殖的抑制及其诱导的HUVEC凋亡, 降低内脂素诱导HUVEC上清液中IL-6及TNF-α的量并能降低HUVEC中p-JNK及Bax蛋白的表达, 增加HUVEC内Bcl-2蛋白表达。结论 小檗碱可减少IL-6及TNF-α的分泌, 减轻内脂素诱导的HUVEC损伤, 其机制可能与抑制JNK信号转导通路有关。

Objective To study the regulation of c-Jun N-terminal kinase (JNK) pathway on interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) secreted in human umbilical vein endothelial cell (HUVEC) induced by visfatin and the intervention of berberine. Methods HUVEC was cultured for the in vitro experiment, visfatin was added at different concentration (0, 50, 100, and 200 μg/L) and in different times (0, 6, 12, 24, and 48 h) to obserbe the effect on HUVEC. MTT assay was used to detect the proliferation of HUVEC, apoptosis rate was measured by Flow Cytometer, the contents of IL-6 and TNF-α in HUVEC supernatant were determined by enzyme-linked immuno-sorbent assay (ELISA) assay, and the protein expressions of p-JNK, Bax, and Bcl-2 in HUVEC lysate were determined by Western blotting. Berberine (50 μmol/L) and SP600125 (10 μmol/L) were added to interfere HUVEC and to detect the effect of berberine. Results Compared with the control group, visfatin (100 μg/L) could significantly decrease the proliferation of HUVEC, induce HUVEC apoptosis, and increase the IL-6 and TNF-α contents in HUVEC supernatant. Meanwhile, it could increase p-JNK and Bax expression, decrease Bcl-2 expression after 24 h. Compared with visfatin group, berberine and SP600125 could increase the proliferation of HUVEC while decrease the IL-6 and TNF-α contents in HUVEC supernatant, restrain p-JNK and Bax expression while increase Bcl-2 expression in HUVEC. Conclusion Berberine could decrease the contents of IL-6 and TNF-α in HUVEC supernatant and ease the injury induced by visfatin, the protective mechanism is related to the inhibition of JNK signal pathway activation.

国家自然科学基金资助项目(81460680, 81373574);江西省科技计划项目(20135BBG70002);广州市海珠区科技计划项目(2013-cg-35)