NF_κBTGF_β1在糖尿病大鼠心肌及主动脉中表达.docVIP

NF_κBTGF_β1在糖尿病大鼠心肌及主动脉中表达 　　中图分类号:R587.1 文献标识 码:A 文章编号:1009_816X(2010)03_0186_02 　　 　　The Expression of NF_κB and TGF_β1 in the Myocardium and Aorta of STZ_inducedDiabetic Rats. ZHOU Ye, QIU Ming_cai, Department of cadre health care, The first peoples hos pital of Hangzhou, Zhejiang 310006, China 　　[Abstract] ObjectiveTo observe the expression of Nuclear factor_κB(NF_κB) a nd Transforming growth factor_β1(TGF_β1) in the myocardium and aorta of diabetic rats induced by STZ.Methods20 male Wister rats were divided into two groups. 10 rats without any treatmen t were served as control group. 10 rats were diabetic rats induced by STZ. At 34weeks, 20 male Wister rats were sacrificed for pathological and Immunohistochem ical study. The pathological changes were assessed by HE and Masson staining. Th e expression of cytokines (NF_κB and TGF_β1) in the myocardium and aorta of di abetic rats was assessed with immunohistochemistry.ResultsIn comparison with c ontrol group, the expressions of NF_κB and TGF_β1 in the myocardium and aortaof diabetic rats induced by STZ were significantly increased (both P1 6.7mmol/L视为造模成功。34周后各组大鼠均股动脉放血处死。迅速取大鼠心肌和主动脉以 10%福尔马林溶液固定。 　　1.2.2 HE、MASSON染色:心肌及主动脉组织做5μm连续切片,按常规程序进行染色。 　　1.2.3 免疫组化检测:所有标本的心肌及主动脉组织切片厚5μm,常规脱蜡至水,微波 抗原修复,滴加封闭血清,心肌组滴加1:150稀释的NF_κB、1:300稀释的TGF_β1一抗; 主动脉组滴加1:50稀释的NF_κB、1:300稀释的TGF_β1一抗,4℃过夜,DAB显色。NF_κB 、TGF_β1均以细胞膜及胞浆内出现棕黄色颗粒为阳性细胞。以积分法半定量描述所取标本 的染色情况,每张切片随机观察10个高倍视野,按染色强度分为4级:0分为阴性,1分为弱 阳性,2分为中度阳性,3分为强阳性。阳性细胞数:无阳性细胞数为0分,阳性细胞数1%～2 5%为1分,阳性细胞数26%～50%为2分,阳性细胞数51%～75%为3分,阳性细胞数76%～100%为 4分,每张切片的染色积分以这二者之和表示。 　　1.3 统计学分析:实验数据以x-±s 表示,采用SPSS11.5软件处理,组间比较用单因素方差分析。 　　 　　2 实验结果 　　 　　2.1 病理学观察:DM大鼠可见心肌细胞肥大,肌纤维的溶解、破坏、肌横纹的消失,玻璃 样变??、间质纤维化,并可见淋巴细胞的浸润。MASSON染色心肌细胞间质和血管周围均可以 看到绿色的胶原纤维组织。DM大鼠的主动脉管壁中可见淋巴细胞的浸润。MASSON染色以绿色 的胶原纤维为主。 　　2.2 免疫组化分析:主动脉:CON组内膜为弱阳性,中膜和外膜阴性。(见图1-A,C)DM组 有大量的NF_κB、TGF_β1的沉积,主要集中在血管内膜上,外膜也有一定的沉积。(见图1 -B,D)。两组表达水平比较有统计学意义,见表1。心肌:CON组为阴性,小部分为弱阳性。 (见图2-E,G)DM组心肌细胞间均可见大量的NF_κB、TGF_β1的沉积。(图2-F,H)。两组 比较结果亦有统计学意义,见表