CAJ | 학술논문

目的组织化学法检测视网膜上caspase-3和血管内皮生长因子(VEGF)的荧光表达.糖尿病鼠建模后2周,对实验组行玻璃体腔注射p38 MAPK抑制剂SB203580,注射后6周检测caspase-3和VEGF的荧光表达,caspase-3免疫荧光染色法计数RGC凋亡数量.采用SPSS 13.0应用软件进行统计分析.结果正常对照组大鼠中,IgG染色局限于血管腔内,几乎没有渗漏的迹象.糖尿病鼠建模后8周,IgG渗漏明显增加.SB203580璃体腔注射6周后的糖尿病鼠IgG渗漏减少明显.荧光免疫组织化学及相对定量分析结果显示,SB203580玻璃体腔注射6周后的糖尿病鼠视网膜上VEGF荧光表达呈下降趋势,VEGF相对荧光量从糖尿病鼠8周时较正常对照组高2.9倍减少到仅高于正常对照组1.8倍,两者比较差异有统计学意义(t=5.203,P<0.01).caspase-3免疫荧光染色法RGC凋亡计数结果显示,SB203580玻璃体腔注射6周后,caspase-3-阳性节细胞凋亡数与未注射SB203580相比明显减少,两者差异有统计学意义(t=5.731,P<0.01).结论 p38 MAPK抑制剂SB203580能够减轻糖尿病早期血视网膜屏障的破坏和视网膜节细胞的调亡,提示p38 MAPK信号通路在糖尿病早期对DR的发展起着一定的作用.
목적 조직화학법검측시망막상caspase-3화혈관내피생장인자(VEGF)적형광표체.당뇨병서건모후2주,대실험조행파리체강주사p38 MAPK억제제SB203580,주사후6주검측caspase-3화VEGF적형광표체,caspase-3면역형광염색법계수RGC조망수량.채용SPSS 13.0응용연건진행통계분석.결과 정상대조조대서중,IgG염색국한우혈관강내,궤호몰유삼루적적상.당뇨병서건모후8주,IgG삼루명현증가.SB203580리체강주사6주후적당뇨병서IgG삼루감소명현.형광면역조직화학급상대정량분석결과현시,SB203580파리체강주사6주후적당뇨병서시망막상VEGF형광표체정하강추세,VEGF상대형광량종당뇨병서8주시교정상대조조고2.9배감소도부고우정상대조조1.8배,량자비교차이유통계학의의(t=5.203,P<0.01).caspase-3면역형광염색법RGC조망계수결과현시,SB203580파리체강주사6주후,caspase-3-양성절세포조망수여미주사SB203580상비명현감소,량자차이유통계학의의(t=5.731,P<0.01).결론 p38 MAPK억제제SB203580능구감경당뇨병조기혈시망막병장적파배화시망막절세포적조망,제시p38 MAPK신호통로재당뇨병조기대DR적발전기착일정적작용.
Objective To investigate the protective effect of blocking the signal path of p38 mitogen-activated protein kinase on blood-retinal barrier (BRB) and retinal ganglion cells (RGC) in early diabetic rats. Methods A total of 60 Wistar rats were divided into the control and diabetes group, with 30 rats in each group. Diabetes was induced in rats in diabetes group by peritoneal injection of streptozotocin (STZ) ; the plasma glucose level of >16.7 mmol/L indicated that the diabetes model was set up successfully. The rats in the control group underwent peritoneal injection of equivalent sodium citrate solution. IgG leakage method was used to measure the damage of BRB function and vascular leakage. The expression and localization of caspase-3 and vascular endothelial growth factor (VEGF) in retina of diabetic rats were examined by immunohistochemistry analyses. Two weeks after the establishment of the diabtes model, the rats in diabtes group underwent intravitreal injection with SB203580, a p38 inhibitor; six weeks after the injection, the expression of caspase-3 and VEGF was detected, and the number of apoptosis RGC was counted via immunofluorescence technique. Results In the contral group, IgG staining located in the blood vessels with little leakage; while the IgG leakage was much more obvious in the diabetes group eight weeks after the establishment of the model. Six weeks after intravitreal injection with SB203580, the leakage decreased in diabtes rats. The results of semi-quantitative analysis and fluorescence immunohistochemistry showed that compared with the results in diabetes rats 8 weeks after intravitreal injection (2.9 times much more than that in the control group), the fluorescence expression of VEGF decreased in diabetes rats six weeks after intravitreal injection (1.8 times much more than that in the control group). The apoptisis RGC number in rats 6 weeks after intravitreal injection of SB203580 was much less than that in rats without intravitreal injection (t=5. 731, P<0. 01). Conclusions SB203580 can alleviate the disruption of BRB and apoptosis of RGC in early diabetes rats, which suggests that p38 MAPK pathways appear to be directly involved in the pathogenesis of early diabetic retinopathy.