| Objective: 1.To observe the expression of heparanase and blood coagulation factor Ⅴ、Ⅶ、Ⅷ、Ⅸ in peripheral serum of patients with retinal vein occlusion, and evaluate the relationship between heparanase, mRNA, and blood coagulation factor Ⅴ、Ⅶ、Ⅷ、Ⅸ in retinal vein occlusion patients.Methods: A non-randomized single-center clinical research methods was adopted. All programs were agreed by medical ethical review committee of the first affiliated hospital of Kunming medical university and abided by Helsinki declaration. All cases were divided into two groups: RVO group and control group. All patients were informed before experiment. All cases were diagnosed of retinal vein occlusion （RVO） by Optical coherence tomography（OCT） and Fundus fluorescein angiography（FFA） in the First Affiliated Hospital of Kunming Medical University from Jan2014 to Jan2015. 40 cases with RVO patients and 40 patients without RVO were divided into two groups:1、The case group:40 case with RVO were divided into two groups.2、The control group:40 case without RVO （simple age-related cataract cases）. All 2ml fasting vein blood samples drawed from case group and the control group were sent to the Clinical laboratory center of the first affiliated hospital of kunming medical university to detect the activity of blood coagulation factor Ⅴ、Ⅶ、Ⅷ、Ⅸ. At the same time,6ml（The citrate anticoagulation） peripheral vein blood collected respectively from the patients in the case group and the control group were separated into serum and blood cell samples by centrifugal machine （1200g,20min,4℃）,,which were placed in -80 degrees freezer（for the experiment of RT-PCR, ELISA and enzyme activity determination）. In the 80 case of serum, the Gene expression of Heparanase were detected by RT-PCR experiment, the expression of Heparanase protein,IL-16,MMP-2,MMP-9,TLR-2 and TLR-4 were detected by ELISA experiment, and the enzyme activity of heparanase were detected by enzyme activity determination method.Results:1.The expression of Heparanase:（1） The level of mRNA: The expression of heparanase mRNA were detected both in RVO group and control group by RT-PCR, After quantitative analysis, The level of heparanase mRNA in RVO group and control group was 3.12±0.50,1.00±0.65 respectively. The level of heparanase mRNA in the RVO group was 3.12 times higher than that in the control group, the difference was statistically significant（P< 0.05）. （2） The level of heparanase protein: heparanase protein were detected both in RVO group and control group by ELISA analysis method, the level of heparanase protein in RVO group and the control group was 3.83±0.70,1.37±0.88 respectively. The level of heparanase protein in the RVO group was 2.79 times higher than that in the control group, the difference was statistically significant （P<0.05）. （3） The results of Enzyme activity test: the expression of enzyme activity were detected both in RVO group and control group, the level of enzyme activity in RVO group and the control group was 0.39±0.11, 0.12±0.13 respectively. The level of enzyme activity in the RVO group is 3.25 times higher than that in the control group, the difference was statistically significant （P＜0.05）.2. The results of coagulation factor Ⅴ、Ⅶ、Ⅷ、Ⅸ、activity tests:（1） Coagulation factor Ⅴ、Ⅶ、Ⅷ、Ⅸ in RVO group is 100.00±17.25、99.50±26.00、199.50±53.75、 137.50±64.00 respectively. in control group is 103.00±27.30、102.00±49.00、 141.00±94.00、88.50±35.00 respectively. There is no significant difference between the two group about coagulation factor Ⅷ （P>0.05）. but the difference of coagulation factor Ⅴ、Ⅶ、Ⅸ in RVO group and control group was statistically significant （P<0.05）.3. The correlatively analysis between Heparanase and coagulation factor Ⅴ, Ⅶ, Ⅷ, Ⅸ in peripheral serum: In RVO group, each P values was 0.347、0.409、0.001、 0.002 respectively. There were not statistically significant between Heparanase and coagulation factor Ⅴ,Ⅶ （P>0.05）, but between Heparanase and coagulation factor Ⅷ, Ⅸ. there were statistically significant（P<0.05）.In control group, each P values was 0.819、0.558、0.974、0.661 respectively. There were not statistically significant （P>0.05）.Conclusion:1.The expression of Heparanase mRNA, Heparanase protein and Heparanase enzyme activity increased in the peripheral serum of patients with RVO, which suggested that Heparanase may play a role in the occurrence of RVO.2. The activity of coagulation factor Ⅷ、Ⅸ increased in the peripheral blood of patients with RVO, which suggested that coagulation factor Ⅷ、Ⅸ may play a role in the occurrence of RVO.3.Heparanase appeared to be positively correlated with coagulation factor Ⅴ、 Ⅶ in RVO. This suggested that Heparanase may play a role in the occurrence of RVO by the way of increasing coagulation?Objective; 1. To observe the expression of IL-6,MCP-1 in aqueous humor of patients with retinal vein occlusion and IL-16,MMP,TLR in peripheral hurum of patients with retinal vein occlusion,and explore the relationship between heparanase and those cytokines.MeUiods: Anon-randomized single-center clinical research methods was adopted. All programs were agreed by medical ethical review committee of the first affiliated hospital of Kunming medical university and abided by Helsinki declaration. All cases were divided into two groups:RVO group and control group. All patients were informed before experiment. All cases were diagnosed of retinal vein occlusion（RVG）by Optical coherence tomography（OCT） and Fundus fluorescein angiography（FFA） in the First Affiliated Hospital of Kunming Medical University from Jan2014 to Jan2015.40 cases with RVO patients and 40 patients without RVO were divided into two groups: 1、The case group: 40 case with RVO were divided into two groups. 2、The control group: 40 case without RVO（ simple age-related cataract cases）,All 2ml fasting vein blood samples drawed from case group and the control group were sent to the Clinical laboratory center of the first affiliated hospital of kunming medical university to detect the activity of blood coagulation factor V、Ⅶ、Ⅷ、IX. At the same time,6ml（The citrate anticoagulation） peripheral vein blood collec1:ed respectively from the patients in the case group and the control group were separated into serum and blood cell samples by centrifiigal machine（1200g,20 min,4°C）,,which were placed in-80 degrees freezer(for the experiment of ELISA. In the 80 case of serum,the expression oflL-16, MMP-2,MMP-9,TLR-2 and TLR-4 were detected by ELISA experiment. In order to know the expressing of IL-6 and MCP-1 in aqueous humor with retinal vein occlusion and macular edema, the 0.2ml aqueous humor of the 18 cases in RVO group and 20 cases in control groups were collected respectively, All aqueous humors samples were placed in-80 degrees freezer and were divided into two groups: The case group: 18 case with RVO. The control group:20 case without RVO.The expression of IL-6 and MCP-1 in aqueous humor of the 38 case were detected by ELISA.Results: 1.The expressions of IL-6、MCP-1 in the in aqueous humor（l）the level of IL-6: The expression of IL-6 were detected both in RVO group and control group by ELISA, After quantitative analysis. The level of IL-6 in RVO group and control group was 316.85±91.25, 77.06±2.41 respectively. The level of BL-6 in the RVO group was higher than that in the control group, the difference was statistically significant（P <0.05）.（2） the level of MCP-1: The expression of MCP-1 were detected both in RVO group and control group by ELISA,After quantitative analysis,The level of MCP-1 in RVO group and control group was 1619.37±815.46, 289.36±11.47 respectively.The level of MCP-1 in the RVO group was higher than that in the control group, the difference was statistically significant（P < 0.05）.2.The expressions of IL-16、MMP-2、MMP-9、TLR-2、TLR-4 in peripheral serum:The expressions of IL-16、MMP-2、MMP-9、TLR-2、TLR-4 were detected both in RVO group and control group by ELISA, After quantitative analysis,The level of IL-16、MMP-2、MMP-9、TLR-2、TLR-4 in RVO group were 896.90±214.64、363.51±9.97、890205.25±254941.30、36..56±8.99、6.87±0.86 respectively. In control group were 420.84±180.72、260.5±8.37、173682.30±146913.06、11.71±9.47、3.61±..83 respectively. The levels of IL-16、MMP-2、MMP-9、TLR2、TLR-4 in the RVO group were higher than that in the control group, the difference were statistically significant（P < 0.05）.3.The correlatively analysis results between Heparanase in peripheral serum and IL-6、MCP-1 in the aqueous humor:（1）In RVO group: the correiative analysis result between Heparanase and IL-6,P value is 0.616, the difference was not statistically significant（P >0.05）. The correlative analysis result between Heparanase and MCP-1,P value is 0.044, the difference was statistically significant（P <0.05）.（2） In control group: the correlatively analysis results between Heparanase and IL-6,P value is 0.016,the differenee was statistically significant（P <0.05）.In RVO group the correlatively analysis results between Heparanase and MCP-1,P value is 0.230, the difference was no statistically significant（P >0.05）.4.In peripheral serum, the corrdatively analysis between Heparanase and IL-16、MMP-2, MMP-9、 TLR-2> TLR-4:（1） In RVO group; the P values of Heparanase and IL-16、MMP-2、MMP-9、TLR-2、TLR-4 are 0.012、0.033、0.014、0.038、0.000 respectively. The difference were statistically significant（P <0.05.）（2）In control group: the values of P of Heparamse and IL-16、MMP-2、MMP-9、TLR-2、TLR-4 are 0.084、0.260、0.936、0.677、0.513 respectively.The difference were no statistically significant（P >0.05）.Condusiont 1. the expression of IL-16、MMP-2、MMP-9、TLR-2、TLR-4 in the peripheral serum blood increased in patients with RVO, which appeared to be positively correlated with Heparanase.lt suggested that Heparanase may play a role in the occurrence of retinal vein occlusion by the way of releasing inflammatory cytokines to involve in vascular lesions and the process of venous thrombosis2. the expression of IL-6、 MCP-1 in the aqueous humor in RVO with macular edema patients It suggested that IL-6 and MCP-1 play a role in the occurrence of retinal vein occlusion with macular edema.MCP-1 appeared to be positively correlated with Heparanase It suggested that MCP-1 may play a role in the occurrence of retinal vein occlxision with macular edema by the way of Heparanase. |
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