CAJ | 학술논문

研究观察了重组人白介素10 (rhIL-10)对晚期糖基化终产物(AGE) 刺激下离体大鼠胸主动脉血管平滑肌细胞增殖及对SD大鼠血管损伤后新生内膜增殖的影响. 体外培养大鼠主动脉血管平滑肌细胞, 采用 MTS/PES法确定血管平滑肌细胞的增殖状态; 应用流式细胞术测定细胞周期; 利用 p44/42 磷酸化抗 MAPK抗体的蛋白免疫印迹法测定p44/42 MAPK磷酸化蛋白表达. 利用大鼠颈动脉血管损伤模型, 观察 rhIL-10对新生内膜增殖的影响.结果显示: (1) AGE处理组与对照组相比, AGE对血管平滑肌细胞增殖具有明显的刺激作用(P<0.05). rhIL-10单独应用对血管平滑肌细胞生长没有影响(P>0.05).在AGE刺激下, 低至100 ng/ml的rhIL-10可抑制血管平滑肌细胞的生长(P<0.05). (2) 流式细胞术测定的结果显示, rhIL-10可以使AGE作用下的VSMC大部分处于G0/G1期, 与对照组相比有明显差异(P<0.01).(3) AGE对p44/p42 MAPK磷酸化蛋白表达有显著的增强作用, 此作用可被rhIL-10抑制(P<0.001). (4) 大鼠颈动脉损伤后, rhIL-10治疗组的动脉血管新生内膜/中层面积比低于对照组约45% (P<0.01).表明抗炎细胞因子rhIL-10可抑制AGE诱导的大鼠血管平滑肌细胞增殖和血管新生内膜的增殖.
연구관찰료중조인백개소10 (rhIL-10)대만기당기화종산물(AGE) 자격하리체대서흉주동맥혈관평활기세포증식급대SD대서혈관손상후신생내막증식적영향. 체외배양대서주동맥혈관평활기세포, 채용 MTS/PES법학정혈관평활기세포적증식상태; 응용류식세포술측정세포주기; 이용 p44/42 린산화항 MAPK항체적단백면역인적법측정p44/42 MAPK린산화단백표체. 이용대서경동맥혈관손상모형, 관찰 rhIL-10대신생내막증식적영향.결과현시: (1) AGE처리조여대조조상비, AGE대혈관평활기세포증식구유명현적자격작용(P<0.05). rhIL-10단독응용대혈관평활기세포생장몰유영향(P>0.05).재AGE자격하, 저지100 ng/ml적rhIL-10가억제혈관평활기세포적생장(P<0.05). (2) 류식세포술측정적결과현시, rhIL-10가이사AGE작용하적VSMC대부분처우G0/G1기, 여대조조상비유명현차이(P<0.01).(3) AGE대p44/p42 MAPK린산화단백표체유현저적증강작용, 차작용가피rhIL-10억제(P<0.001). (4) 대서경동맥손상후, rhIL-10치료조적동맥혈관신생내막/중층면적비저우대조조약45% (P<0.01).표명항염세포인자rhIL-10가억제AGE유도적대서혈관평활기세포증식화혈관신생내막적증식.
The purposes of this study was to determine the effects of recombinant human interleukin-10 (rhIL-10) on proliferation of vascular smooth muscle cells (VSMCs) stimulated by advanced glycation end products (AGE) and neointima hyperplasia after rat carotid arterial injury. Rat aortic VSMCs were cultured and treated with rhIL-10 or AGE respectively, and then co-treated with rhIL-10 and AGE. Proliferation of VSMCs was quantified by colormetric assay. Cell cycle analysis was performed by flow cytomertry. Sprague-Dawley rats were treated with recombinant human IL-10 (rhIL-10) for 3 d after carotid arteries injury. The ratio of neointima to media area at the site of arterial injury was measured 28 d after balloon injury. The p44/42 MAPK activity was evaluated by the immunoblotting technique using anti-p44/42 phospho-MAPK antibody. Compared to control, AGE stimulated VSMCs proliferation. rhIL-10 alone had no effect on VSMCs growth. With AGE stimulation, rhIL-10, at dose as low as 10 ng/ml, inhibited VSMCs growth (P<0.05). The cell number in G0/G1 phase of AGE and rhIL-10 co-treatment group was higher than that of AGE treatment alone (P<0.01) by flow cytometry analysis. Compared with the control group of neointima hyperplasia in rats, the ratio of neointima to media area of recombinant human IL-10 group was reduced by 45% (P<0.01). The p44/42 MAPK activity was significantly enhanced by AGE. The AGE effects were opposed by rhIL-10. The anti-inflammatory cytokine rhIL-10 inhibits AGE-induced VSMCs proliferation. Recombinant human IL-10 also inhibited neointima hyperplasia after carotid artery injury in rats. The results suggest the possibility that recombinant human IL-10, as a potential therapeutic approach, prevents neointimal hyperplasia.