组织激肽释放酶在糖尿病视网膜病变患者血清中的变化及其调节血管新生机制

目的　检测组织激肽释放酶（ｔｉｓｓｕｅｋａｌｌｉｋｒｅｉｎ，ＴＫＬＫ）、血管内皮细胞生长因子（ｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｇｒｏｗｔｈｆａｃｔｏｒ，ＶＥＧＦ）和可溶性细胞间黏附分子-１（ｓｏｌｕｂｌｅｉｎｔｒａｃｅｌｌｕｌａｒａｄｈｅｎｓｉｏｎｍｏｌｅｃｕｌ-１，ｓＩＣＡＭ-１）在糖尿病视网膜病变（ｄｉａｂｅｔｉｃｒｅｔｉｎｏｐａｔｈｙ，ＤＲ）患者血清中的变化，观察ＴＫＬＫ在低氧条件下对人视网膜微血管内皮细胞（ｈｕｍａｎｒｅｔｉｎａｌｍｉｃｒｏｖａｓｃｕｌａｒｅｎｄｏｔｈｅｌｉａｌｃｅｌｌｓ，ＨＲＭＥＣｓ）中ＶＥＧＦ和ＩＣＡＭ-１表达的影响。方法　收集２型糖尿病患者６０例，按照ＤＲ分期标准将患者分为糖尿病无ＤＲ组（ＤＭ组）、非增生性ＤＲ组（ＮＰＤＲ组）和增生性ＤＲ组（ＰＤＲ组），收集同期在本院体检的志愿者作为对照组，每组２０例。ＥＬＩＳＡ法检测血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１的水平。体外ＨＲＭＥＣｓ分别进行常氧和低氧培养，不同浓度重组ＴＫＬＫ处理后，检测细胞增殖、凋亡以及ＶＥＧＦ和ＩＣＡＭ-１的表达。结果　ＤＭ、ＮＰＤＲ和ＰＤＲ组患者血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１水平明显高于对照组，４组总体差异有统计学意义（Ｆ＝２８．８０５，Ｐ＝０．００２；Ｆ＝３２．０４１，Ｐ＝０．００２；Ｆ＝２６．１６９，Ｐ＝０．００１）；ＰＤＲ患者血清中ＴＫＬＫ、ＶＥＧＦ和ｓＩＣＡＭ-１水平显著高于ＤＭ和ＮＰＤＲ组（均为Ｐ＜０．００１）。ＴＫＬＫ与ＶＥＧＦ（ｒ＝０．６２３，Ｐ＜０．０１）和ｓＩＣＡＭ-１水平（ｒ＝０．５９８，Ｐ＜０．０１）均呈正相关。１０μｇ?ｍＬ－１ｒｈＴＫＬＫ可显著抑制低氧诱导的ＨＲＭＥＣｓ增殖以及ＶＥＧＦ和ＩＣＡＭ-１的表达，并可促进细胞凋亡（Ｐ＜０．０５）。结论　ＴＫＬＫ通过与ＶＥＧＦ和ＩＣＡＭ-１相互作用而影响ＤＲ的进展。

Changes of tissue kallikrein in serum of patients with diabetic retinopathy and its role in retinal angiogensis

Objective To determine the levels of serum tissue kallikrein ( TKLK) ,vascular endothelial growth factor ( VEGF) and soluble intracellular adhension molecul-l ( sICAM-I) in diabetic retinopathy ( DR) patients.and evaluate the effects of TKLK on the expression of VEGF and ICAM-I in human retinal vascular endothelial cells ( HRMECs) under hypoxia. Methods Sixty DM patients were enrolled and grouped according to the staging criteria of DR into: DM patients without DR group ( DM group) ,non-proliferative DR group ( NPDR group) and proliferative DR group ( PDR group ) . Simultaneously ,20 healthy volunteers were collected as the controls. ELISA was applied to measure the levels of serum TKLK .VEGF and sICAM-I in all the eligible subjects,and the correlations between them were analyzed. HRMECs were incubated in vitro with various concentrations of thTKLK. and then were grown in normoxia and hypoixa conditions. Cell viability and apoptosis as well as the expression of VEGF and ICAM-I were examined. Results The serum levels of TKLK.VEGF and sICAM-I in DM .NPDR and PDR groups were higher than that in control group , there were significant differences among four groups ( F = 28. 805 .P = 0. 002 ; F = 32. 041 ,P = 0. 002 .F = 26. 169 .P =0. 001 ). Meanwhile , the serum levels of TKLK.VEGF and sICAM-I in PDR group were higher than those in DM and NPDR groups ( all P < 0. 001 ) . There were significant positive correlations between the levels of TKLK and VEGF ( r = 0. 623 ,P < 0. 01 ) as well as TKLK and sICAM-I ( r = 0. 598 .P < 0. 01 ) . 10 Vg . mL -l thTKLK dramatically inhibited the hypoxia-induced proliferation of HRMECs and expression of VEGF and ICAM-I .while significantly promoted cell apoptosis (P < 0. 05 ) . Conclusion TKLK participate in the progression of DR via interaction with VEGF and ICAM-I.