塞来昔布对嘌呤氨基学核苷诱导的足突细胞凋亡影响的实验研究-内科学专业毕业论文.docxVIP

江苏大学硕士论文 Caspase．3活性检测试剂盒检澳lJCaspase．3蛋白酶活性水平。在24h，用 Western blot技术检测各组P53及COX．2水平。 结果 1．在33℃增殖状态下，足细胞呈鹅卵石状，在37℃分化状态下 的足细胞呈分叉状。正常足细胞在33℃表达COX．2，在37℃分化14d 后，用低血清培养液培养后，随着时间延长，COX．2表达逐渐增多， 24h达最高，以后再逐渐减少。 2．与正常对照组相比较，DEX、CELE组凋亡率无明显变化 (P0．05)，PA组凋亡率明显增高(P0．001)，DEX+PA、CELE+PA 组凋亡率较PA组明显下降(P0．05)。 3．各组Caspase一3活性无明显差异(P0．05)。 4．24h时，PA组P53表达较正常对照组明显增多，用DEX、CELE 干预后其表达明显降低(P0．05)。 5．正常对照组即可表达COX．2，用PA诱导24h，COX．2表达明显 增加，用DEX、CELE干预后COX．2表达水平有所减少(P0．05)。 结论 1．COX．2在足细胞上的表达呈时间依赖性。 2．塞来昔布可以抑制PA诱导的足细胞凋亡，P53参与了PA诱导 的足细胞凋亡过程。塞来昔布抑制PA诱导的足细胞凋亡可能与足细 胞所表达的COX．2水平有一定的相关性。 关键词：塞来昔布，足细胞，细胞凋亡，环加氧酶2 II 江苏大学硕士论文 ABSTRACT Background The glomerular visceral epithelial cell，also called as podocyte，is a hi曲ly specialized and terminally differentiated cell which plays a major role in establishing the glomerular filtration barrier．Podocytes therefore have limited ability to be replaced if lost．There is a growing body of literature showing the reduced podocyte number is a critical determinant underlying the development of glomerulosclerosis．Decreased podocyte number is caused by detachment of cells from the glomerular basement membrane and／or by apoptosis．The underlying mechanisms of which the specific cyclooxygenase一2(cox一2)inhibitors have been shown to decrease proteinuria and retard progressive glomerular injury in some animal models have not been addressed formally．Accordingly,in this study we investigated the direct effect of celecoxib，a specific COX一2 inhibitors，on podocyte apoptosis induced by puromycin aminonucleoside (PA)and the mechanisms that underlie this effect in order to further search for clinical therapeutic target on chronic progressive glomerular lnJury． Methods 1．Experiments were performed using early-passage growth-restricted， conditionally immortalized mouse podocytes．The expression of COX一2 III 江苏大学硕士论文 in podocytes was measured by use of Western blot analysis．The effects of specific COX一2 inhibitors celecoxib on podocyte apoptosis induced by PA will be examined on