CAJ | 학술논문

目的研究高血糖对人巨噬细胞中抑瘤素M表达的影响以及抑瘤素M对人主动脉平滑肌细胞增殖的影响.方法采用佛波酯孵育人单核/巨噬细胞系THP-1细胞,诱导其分化为巨噬细胞.应用人细胞因子抗体芯片筛选出受高血糖(15 mmol/L)调控的人巨噬细胞源性细胞因子,并经Western blot和酶联免疫吸附试验(ELISA)技术验证.运用逆转录-聚合酶链反应(RT-PCR)明确人主动脉内皮细胞、平滑肌细胞及巨噬细胞中是否存在抑瘤素M受体GP130-OSMRβ二聚体及GP130-LIFR二聚体表达.运用细胞增殖实验(XTT法)研究抑瘤素M对人主动脉平滑肌细胞增殖的影响.采用单因素方差分析进行统计学分析.结果佛波酯孵育48 h可诱导THP-1细胞分化为巨噬细胞.细胞因子芯片结果显示高血糖可上调抑瘤素M在巨噬细胞中的表达,并为Western blot及ELISA所证实.RT-PCR显示人主动脉内皮细胞、平滑肌细胞及巨噬细胞均可表达GP130-OSMRβ二聚体及GP130-LIFR二聚体.XTT法证实抑瘤素M可促进血管平滑肌细胞增殖,与0μg/L抑瘤素M(吸光度为0.468±0.032)相比,5μg/L(吸光度为0.503±0.026)、10μg/L(吸光度为0.520±0.027)、20μg/L抑瘤素M(吸光度为0.513±0.026)人主动脉平滑肌细胞增殖均明显增加(F=10.40,P<0.05).结论高血糖可促进巨噬细胞中抑瘤素M的表达和分泌.抑瘤素M通过与人主动脉平滑肌细胞中的受体结合而促进血管平滑肌细胞的增殖,进而部分参与糖尿病大血管病变的发生发展.
목적 연구고혈당대인거서세포중억류소M표체적영향이급억류소M대인주동맥평활기세포증식적영향.방법 채용불파지부육인단핵/거서세포계THP-1세포,유도기분화위거서세포.응용인세포인자항체심편사선출수고혈당(15 mmol/L)조공적인거서세포원성세포인자,병경Western blot화매련면역흡부시험(ELISA)기술험증.운용역전록-취합매련반응(RT-PCR)명학인주동맥내피세포、평활기세포급거서세포중시부존재억류소M수체GP130-OSMRβ이취체급GP130-LIFR이취체표체.운용세포증식실험(XTT법)연구억류소M대인주동맥평활기세포증식적영향.채용단인소방차분석진행통계학분석.결과 불파지부육48 h가유도THP-1세포분화위거서세포.세포인자심편결과현시고혈당가상조억류소M재거서세포중적표체,병위Western blot급ELISA소증실.RT-PCR현시인주동맥내피세포、평활기세포급거서세포균가표체GP130-OSMRβ이취체급GP130-LIFR이취체.XTT법증실억류소M가촉진혈관평활기세포증식,여0μg/L억류소M(흡광도위0.468±0.032)상비,5μg/L(흡광도위0.503±0.026)、10μg/L(흡광도위0.520±0.027)、20μg/L억류소M(흡광도위0.513±0.026)인주동맥평활기세포증식균명현증가(F=10.40,P<0.05).결론 고혈당가촉진거서세포중억류소M적표체화분비.억류소M통과여인주동맥평활기세포중적수체결합이촉진혈관평활기세포적증식,진이부분삼여당뇨병대혈관병변적발생발전.
Objective To study the effects of high blood glucose on expression of oncostatin M (OSM) in macrophages and the association of OSM with proliferation of human aortic smooth muscle cells. Methods Monocytic cell line THP-1 was incubated with PMA to induce macrophages. The human macrophage cell line-derived cytokines influenced by hyperglycemia (15 mmol/L) were screened by human cytokine antibody array, which was further confirmed by Western blot and ELISA. RT-PCR was used to determine the expression of OSM receptors GP130-OSMRβ and GP130-LIFR in human aortic endothelial cells, smooth muscle cells and macrophages. The effects of OSM on proliferation of human aortic smooth muscle cell was assessed using the XTT assay. One-way analysis of variances was used for data analysis. Results Incubation with PMA induced mature macrophages. Human cytokine antibody array showed that high blood glucose up-regulated the expression of OSM in macrophages, which was further confirmed by Western blot and ELISA. The expression of GPI30-OSMRβ and GP130-LIFR was found in human aortic endothelial cells, smooth muscle cells and macrophages. XTT assay showed that OSM promoted proliferation of human aortic smooth muscle cells. Compared with 0 μg/L OSM (OD = 0.468 ±0.032), the proliferation of human aortic smooth muscle cells was significantly increased in 5, 10, 20 μg/L OSM (OD = 0.503±0.026, 0.520±0.027, 0.513±0.026, respectively) (F=10.40,P<0.05) Conclusions High blood glucose could increase the expression and secretion of OSM in macrophages. OSM may promote the proliferation of human aortic smooth muscle cells by binding with its receptors, which partly involves in the development of diabetic macroangiopathy.