中国人VEGF基因多态性与DR的关系

目的:探讨血管内皮生长因子(vascular endothelial growth factor,VEGF)I/D基因多态性与糖尿病视网膜病变(diabetic retinopathy,DR)的关系。方法:应用PCR-RFLP方法检测91例2型糖尿病患者和30例对照者的VEGF I/D基因和VEGF水平。结果:NPDR组和PDR组DD基因型频率显著高于NDR组和对照组(P<0.01);NPDR组和PDR组血清VEGF水平显著高于NDR组和对照组(P<0.01)。结论:VEGF I/D基因多态性很可能与DR的发生发展有关,D等位基因可能是DR的易感基因。     

海南黎族2型糖尿病患者VEGF基因多态性与糖尿病视网膜病变关系

目的:探讨海南黎族2型糖尿病患者血管内皮细胞生长因子(VEGF)基因rs2010963和rs3025039的单核苷酸多态性与糖尿病视网膜病变(DR)的相关性。方法:前瞻性研究。随机收集2016-09/2019-10海南黎族2型糖尿病患者89例,其中非增殖期DR(NPDR)患者30例、增殖期DR(PDR)患者33例,2型糖尿病无视网膜病变(DWR)患者26例作为对照。运用聚合酶链式反应-限制性片段长度多态性技术测序法,进行VEGF基因相应位点的多态性分析,比较基因型和等位基因频率的差别。结果:在rs2010963位点上,与DWR组比较,PDR组CC基因型显著升高(P<0.016667),PDR组CG基因型显著降低(P<0.016667),三组患者的GG基因型和C、G等位基因分布比较均无差异(P>0.05)。在rs3025039位点上,DWR组,NPDR和PDR组三组的CC、CT基因型及C、T等位基因分布均无差异(均P>0.05)。海南黎族PDR患者尿素、肌酐水平较DWR和NPDR组均显著升高(均P<0.05)。结论:海南黎族2型糖尿病患者VEGF基因rs2010963基因多态性与DR发生有关,CC基因型可能是DR发生的遗传危险因素,增加PDR的易感性。rs2010963基因多态性对于PDR具有协同作用。     

2型糖尿病视网膜病变严重程度与血清生化全套指标的相关性

目的:分析不同程度2型糖尿病视网膜病变的血清生化全套指标,探讨各生化指标与糖尿病性视网膜病变(diabetic retinopathy,DR)程度的相关性。方法:回顾性收集2018年5月至2020年10月在福建医科大学附属第二医院就诊的2型糖尿病患者。根据眼底情况分为3组:糖尿病不伴视网膜病变(non-diabetic retinopathy,NDR)组(17例)、糖尿病伴非增殖性视网膜病变(non-proliferative diabetic retinopathy,NPDR)组(29例)、糖尿病伴增殖期视网膜病变(proliferative diabetic retinopathy,PDR)组(34例)。采用SPSS 24.0分析比较3组血清生化全套各指标水平,对差异指标与DR相关性采用Pearson相关分析与多重线性回归分析。结果:3组年龄差异无统计学意义(P>0.05);3组的总蛋白、白蛋白、白球比、钙浓度差异均有统计学意义(均P<0.05)。组间比较发现NPDR和PDR组总蛋白、白蛋白、白球比、钙浓度低于NDR组,差异有统计学意义(P<0.05)。Pearson相关分析显示总蛋白(r=?0.290)、白蛋白(r=?0.304),钙浓度(r=?0.252)与DR呈负相关(均P<0.05),多重线性回归提示总蛋白、白蛋白及血钙浓度是DR保护因素(P<0.05)。结论:血清指标总蛋白、白蛋白及血钙浓度与DR成负相关,在生理范围内将这些指标维持相对较高的水平,或许可以为糖尿病视网膜病变的防治提供新思路。     

2型糖尿病患者干眼与糖尿病视网膜病变的相关分析

目的：：调查2型糖尿病患者干眼与不同程度糖尿病视网膜病变( diabetic retinopathy,DR)及黄斑水肿之间的关系。方法：采用横断面研究。选取340例340眼2型糖尿病患者,收集临床资料,分别检测泪河高度、泪液分泌试验(Schirmer Ⅰ test)、泪膜破裂时间(break-up time,BUT)、角膜荧光素染色。所有患者散瞳检查视网膜,评估DR程度及有无临床意义的黄斑水肿。结果：所有患者中,干眼患病率为49.41%。干眼患者的糖尿病病程为11.15±7.07a,无干眼患者的病程为6.92±5.45a,两者之间的差异具有显著统计学意义(P<0.01)。干眼与DR各分期具有明显关系,轻度非增殖性糖尿病视网膜病变( nonproliferative diabetic retinopathy, NPDR )、中度 NPDR、重度 NPDR 和增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)患者相对于无 DR患者的干眼发生可能性分别为1.097倍、1.724倍、2.86倍和5.43倍。黄斑水肿患者较无黄斑水肿患者的干眼发生可能性增加到3.697倍。结论：2型糖尿病患者常常伴发干眼。随着DR的发生及进展,罹患干眼的机会逐步增加。     

不同程度糖尿病视网膜病变干眼症患者波前像差的研究

目的：比较不同程度糖尿病视网膜病变(diabetic retinopathy,DR)干眼症患者波前像差的变化,探讨糖尿病视网膜病变干眼症患者视觉质量下降的原因。

方法：随机选取本院就诊的不同程度糖尿病视网膜病变干眼症患者各40眼,正常对照组40眼。采用Topcon KR-1W视觉质量分析仪记录瞳孔直径4mm和6mm的角膜总高阶像差、彗差、球差及三叶草像差的均方值。采用方差分析比较不同程度糖尿病视网膜病变干眼症患者波前像差及与对照组像差值。

结果：4mm和6mm瞳孔直径下无明显糖尿病视网膜病变(nondiabetic retinopathy,NDR)干眼症组、非增殖性糖尿病视网膜病变(nonproliferative diabetic retinopathy,NPDR)干眼症组及增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)干眼症组较对照组眼总高阶像差、彗差及三叶草像差均有明显升高(P<0.01),且PDR干眼症组较NDR干眼症组和NPDR干眼症组总高阶像差、球差、彗差及三叶草像差均有明显升高(P<0.01)。

结论：不同程度糖尿病视网膜病变干眼与波前像差的增高密切相关,波前像差增高可能是降低糖尿病视网膜病变干眼症患者视觉质量的原因之一,为糖尿病干眼视功能下降提供了依据。     

Nrf2基因多态性与糖尿病视网膜病变的相关性研究

目的：探讨陕西地区汉族人群中,核因子E2相关因子2(Nrf2)基因多态性与糖尿病视网膜病变(DR)的相关性。

方法：收集2016-01/2017-01在我院治疗的386例2型糖尿病(T2DM)患者分为无视网膜病变(DWR)组181例,增生期DR(PDR)组62例和非增生期DR(NPDR)组143例。另外,收集120例非糖尿病且无视网膜疾病的患者作为对照组。采用聚合酶链反应(PCR)联合DNA直接测序法检测Nrf2基因启动子rs6721961位点单核苷酸多态性。

结果： DWR组与对照组比较,rs6721961位点基因型和等位基因分布频率均无差异(P>0.05); PDR组和NPDR组分别与对照组和DWR组比较,突变纯合子(AA基因型)和突变基因(A等位基因)分布频率差异均有统计学意义(P<0.05)。PDR组和NPDR组比较,基因型和等位基因分布频率均无差异(P>0.05)。Logistic多因素回归分析结果显示rs6721961位点A等位基因与DR发生相关(OR=1.532, 95%CI ：1.169～2.008, P=0.014)。

结论： Nrf2基因多态性可能与DR遗传易感性相关。     

2型糖尿病视网膜病变血清C肽测定及临床意义

目的 探讨血清C肽与2型糖尿病视网膜病变(DR)之间的关系及其临床意义.方法 将2型糖尿病住院病人167例分为3组,其中无糖尿病视网膜病变组(NDR组)60例,非增殖性糖尿病视网膜病变组(NPDR组)76例,增殖性糖尿病视网膜病变组(PDR组)31例.采用化学发光免疫分析法分别测定其空腹血清C肽含量并记录其他可能影响糖尿病视网膜病变发生、发展的因素.根据所测血清C肽值重新将病人分为C肽低值组(LCP组)37例,C肽正常值组(NCP组)78例,C肽高值组(HCP组)52例.结果 NDR组、NPDR组、PDR组空腹血清C肽平均水平呈降低趋势,三组之间空腹血清C肽平均水平比较差异有统计学意义(P＜0.01).LCP组、NCP组、HCP组之间DR发生率比较差异有统计学意义(x2=46.702,P＜0.01).Logistic回归分析显示空腹血清C肽水平与DR有明显相关性(B=-1.415,P=0.000),病程与DR有明显相关性(B=0.404,P=0.000).C肽水平和糖尿病病程呈负相关(r=-0.479,P=0.000),C肽水平与DR的程度呈负相关(r=-0.654,P=0.000).结论 血清C肽在2型DR的发生、发展过程中可能有重要的病理意义,其水平降低可能是2型糖尿病患者DR进展的临床标志.

Abstract：

Objective To explore the relationship of serum C-peptide with diabetic retinopathy (DR)and its clinical significance. Methods A total of 167 inpatients with type 2 diabetes were separated into three groups, NDR group (n=60 examination of ocular fundus showed normal), NPDR group (n=76 examination of ocular fundus showed nonproliferative diabetic retinopathy), PDR group (n=31 examination of ocular fundus showed proliferative diabetic retinopathy). Their fasting serum C-peptide (FCP) was tested with chemiluminescent immunoassay. At the same time, other factors that might affect the onset and progression of DR were recorded. Then all the patients were divided according to their C-peptide fasting levels in quartiles again, low C-peptide group (LCP) [n=37], normal C-peptide group (NCP) [n=78], high C-peptide group (HCP) [n=52].Results In three groups, the mean value of FCP in PDR group was the lowest, the NPDR group had the lower concentration than NDR group, [(0.61±0.33) ng/ml vs (1.34± 0.68) ng/ml vs (2.11 ± 0.80) ng/ml, P ＜0.01]. The incidence of DR among LCP group, NCP group and HCP group showed a significant difference (x2=46.702, P ＜0.01). Logistic regression analysis showed that only C-peptide (B=-1.415, P=0.000), duration of diabetes (B=0.404, P =0.000) were independently associated with DR. A negative correlation was evident between C-peptide levels and duration of diabetes (r=-0.479, P =0.000), a negative correlation was also found between C-peptide levels and DR (r=-0.654, P =0.000). Conclusions Serum C-peptide may have important significance in pathological form of Type 2 DR. The decrease of C-peptide levels may be a maker of the development of DR in type 2 diabetes.     

IL-6基因启动子区-572C/G多态性与大理白族2型糖尿病视网膜病变的关系

目的：探讨IL-6基因启动子区-572C/G多态性与大理白族2型糖尿病(type diabetic mellitus,T2DM)视网膜病变(diabetic retinopathy,DR)人群的相关性及视网膜病变程度的关系。

方法：采用聚合酶链反应联合限制性片段长度多态性分析(polymerase chain reaction-restriction fragment polymorphisms assay,PCR-RFLP)技术及基因测序技术,在大理白族人群中对150例T2DM患者及100例健康对照者(CON组)的IL-6-572C/G位点基因多态性进行检测。150例T2DM患者中,NDR(未合并DR)组57例,NPDR(合并非增生性糖尿病视网膜病变)组77例,PDR(合并增生性糖尿病视网膜病变)组16例。比较各组间基因型及等位基因分布频率,同时收集临床生化指标,最终数据使用SPSS22.0统计软件进行统计学分析。

结果：IL-6基因-572C/G位点组间基因型及等位基因频率相比较,差异均具有统计学意义(P<0.05); 携带C等位基因的个体避免患T2DM的风险为G等位基因的1.182倍(95% CI：1.059～1.319,P=0.004); 携带G等位基因的T2DM患者并发DR的风险为C等位基因的1.667倍(95% CI：1.195～2.326,P=0.003),但基因型及等位基因频率在PDR组与NPDR组比较,差异均无统计学意义(P>0.05); T2DM、NPDR+PDR组与CON组在空腹血糖、甘油三酯、体质量指数比较均有统计学差异(P<0.05); NPDR组与PDR组相比只有空腹血糖比较差异有统计学意义(P<0.05); 合并高血压的个体患T2DM的风险为未患高血压的 3.730倍(95% CI：2.060～6.754,P=0.000),同时患DR的风险为3.997倍(95% CI：2.099～7.612,P=0.000); 不同基因型间临床生化指标比较均无统计学差异(P>0.05)。

结论：IL-6基因-572C/G基因多态性与大理白族T2DM及DR的易感性相关,但临床生化指标对此易感关联没有协同作用,G等位基因是DR和T2DM发病的危险因素,但在NPDR进展至PDR过程中无意义,C等位基因对于T2DM和DR的发病具有保护作用; 高血压、空腹血糖、甘油三酯、体质量指数促进T2DM和DR的发病,空腹血糖在DR病情进展中具有重要作用。     

糖尿病视网膜病变荧光造影临床分析

目的:探讨糖尿病视网膜病变(diabetic retinopathy,DR)眼底镜及眼底荧光血管造影(fundus fluorescence angiography,FFA)检查表现的差异,评价FFA在DR早期诊断中的价值.方法:经我院内分泌科确诊为糖尿病的患者分别行散瞳眼底镜检查及FFA检查,行DR临床分期,分析总结两者的表现差异.结果:120例(238眼)糖尿病患者中,眼底镜下"正常眼底"42眼中FFA修正诊断为非增殖性糖尿病视网膜病变(nonproliferative diabetic retinopathy,NDR)Ⅰ期者11眼,修正诊断为NDRⅢ期者3眼;眼底镜下NDR176眼中FFA修正诊断为NDRⅢ期者18眼,修正诊断为增殖性糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)Ⅳ期者18眼;PDR 20眼均未行修正诊断.结论:FFA可早期发现糖尿病视网膜病变的微血管异常,更准确对DR进行诊断及分期.     

对糖尿病视网膜病变患者视网膜血管循环时间的观察

目的：观察不同分期糖尿病性视网膜病变(diabetic retinopathy,DR)视网膜动、静脉循环时间的差异性,分析各期DR视网膜血管循环时间的变化特点。

方法：收集2016-03/2017-03我院眼科确诊的DR患者60例60眼,其中轻中度非增殖期糖尿病视网膜病变(nonproliferative diabetic retinopathy,NPDR)组20例20眼、重度NPDR组20例20眼、增殖期糖尿病视网膜病变(proliferative diabetic retinopathy,PDR)组20例20眼。通过眼底荧光血管造影(undus fluorescein angiography,FFA)检查记录视网膜动脉4个分支主干刚开始显影即臂视网膜循环时间(A1)和完全充盈的时间(A2)、视网膜静脉4个分支主干全部出现层流的时间(V1)和完全充盈的时间(V2),并进行相关分析。

结果：视网膜动脉通过时间(A2-A1)三组比较,差异无统计学意义(F=1.642,P=0.157)。视网膜毛细血管通过时间(V1-A2)、视网膜静脉通过时间(V2-V1)、视网膜动静脉通过时间(V2-A1)三组分别比较,差异均有统计学意义(F=5.794、5.180、5.564,P=0.007、0.009、0.008),其中PDR组均较轻中度和重度NPDR组明显延长,重度NPDR组均较轻中度NPDR组明显延长。

结论： DR病程越长,程度越重,视网膜微血管破坏越严重,视网膜微循环时间越长。     

闭合漏斗型视网膜脱离手术治疗的探讨

目的 探讨闭合漏斗型视网膜脱离合并严重PVR病例的手术治疗方法。方法 对12例进行闭合式三切口玻璃体切除及视网膜复位术。结果 12例视网膜全部复位,7例有不同程度的视功能改善,保持眼球外观。眼轴缩短呈塌陷状外观的病例术后外观及视力均有改善。结论 对闭合漏斗型视网膜脱离合并严重PVR病例手术治疗可以使部分病人保持视功能甚至恢复一定的视力。即使是无望恢复视力者也可以改善眼球的外观。     

血清和房水中miR-377-3p和miR-365-3p表达水平与糖尿病性黄斑水肿程度的相关性

目的:探讨血清和房水中miR-377-3p和miR-365-3p表达水平与糖尿病性黄斑水肿(DME)程度的相关性。方法:前瞻性研究。选取2021-02/2022-02三六三医院收治的初诊为DME患者60例60眼(如双眼发病取严重眼入组,若两眼程度一致取右眼),其中轻度24眼,中度21眼,重度15眼;另选同期本院收治的未发生DME的2型糖尿病患者60例60眼作为对照组。收集所有患者基本临床资料,包括BMI、吸烟史、饮酒史、高血压、高血脂、糖尿病病程、糖化血红蛋白、空腹血糖、Hcy;实时荧光定量PCR(qRT-PCR)法检测血清和房水中miR-377-3p和miR-365-3p表达水平。结果:DME组患者糖尿病病程、糖化血红蛋白、空腹血糖、同型半胱氨酸(Hcy)均显著高于对照组(均P<0.05);DME组患者血清中miR-377-3p和miR-365-3p表达水平均低于对照组(均P<0.05);重度组患者血清和房水中miR-377-3p和miR-365-3p表达水平显著低于中度组和轻度组,中度组显著低于轻度组(均P<0.05);血清中miR-377-3p和miR-365-...     

FOXO3和IL-2在干眼患者结膜上皮细胞及泪液中表达及意义

目的：探究叉头框蛋白O3(FOXO3)、白细胞介素-2(IL-2)在干眼(DE)患者结膜上皮细胞及泪液中的表达及意义。

方法：前瞻性研究。选择2019-03/2021-03收治的DE患者106例，另选同期85位健康体检者为对照。利用实时荧光定量PCR(qRT-PCR)法检测结膜上皮细胞及泪液中FOXO3水平； 酶联免疫吸附(ELISA)法检测样本中IL-2水平； 分析DE患者治疗前后的泪膜破裂时间(BUT)、泪液分泌试验(SⅠt)、角膜荧光素染色(CFS)等眼表临床指标变化； Pearson相关性分析DE患者结膜上皮细胞、泪液中FOXO3和IL-2水平的相关性以及二者与临床指标的关系。

结果：与对照组相比，DE组患者结膜上皮细胞及泪液中FOXO3水平均明显降低，IL-2水平均明显升高(均P<0.01)。与治疗前相比，DE患者治疗后结膜上皮细胞及泪液中FOXO3水平明显上调，IL-2水平明显下调(均P<0.01)。Pearson相关性分析结果显示，结膜上皮细胞、泪液中FOXO3和IL-2水平均呈显著负相关(r=-0.531、-0.469，均P<0.01)。DE患者治疗后BUT、SⅠt指标较治疗前上升，CFS指标下降(均P<0.01)。DE患者结膜上皮细胞FOXO3水平与BUT、SⅠt均呈正相关(r=0.431、0.457，均P<0.01)，与CFS呈负相(r=-0.469，P<0.01)，IL-2水平与BUT、SⅠt均呈负相关(r=-0.416、-0.447，均P<0.01)，与CFS呈正相关(r=0.424，P<0.01)； 泪液FOXO3与BUT、SⅠt呈正相关(r=0.421、0.443，均P<0.01)，与CFS呈负相关(r=-0.474，P<0.01)，IL-2与BUT、SⅠt呈负相关(r=-0.408、-0.429，均P<0.01)，与CFS呈正相关(r=0.419，P<0.01)。

结论：DE患者结膜上皮细胞及泪液中FOXO3水平降低，IL-2水平升高，二者与患者眼表指标存在密切的联系，有望成为DE临床监测和预后评估的实验室辅助指标。     

Effects of Qingkailing on Experimental Allergic Uveitis in Rabbits

Experimental allergic uveitis(EAU)in rabbits was induced by singleintraocular injection of bovine serum albumin(BSA).The average of protein con-centration in aqueous humor of untreated group of rabbits was 14.33±1.21mg/mland the count of tritiated thymidine(~3H-TdR)incorporated into lymphocyte T was3,987±1,156cpm/10~6.The specific antibody responses to BSA in the serum andthe aqueous were 0.508±0.034 and 0.369±0.019(OD)respectively.Meanwhile,the effect of Qingkailing on EAU was observed in comparison withdexamethasone as the positive control.The inflammatory reactions of rabbitstreated with Qingkailing were remarkably diminished clinically and pathological-ly.The amount of protein concentration in aqueous of the treated rabbits was5.79±2.19mg/ml(P<0.01)and the count of ~3H-TdR incorporation was2,455±981 cpm/10~6(P<0.05).The responses of antibody in the serum and theaqueous were 0.414±0.059 and 0.253±0.041 (P<0.01)respectively.Our obser-vations indicate that Qingkailing has a positive therapeutic effect on EAU in therabbits and sustain the suggestion that this drug is capable of suppressing im-mune and phlogistic reactions.Eye Science 1992;8:138-142.     

Wegener’s Granulomatosis of the Orbit: A Review of Clinical Features and Updates in Diagnosis and Treatment

Wegener’s Granulomatosis (WG) is an uncommon systemic vasculitic disease involving the respiratory system and kidneys. It is the most common antineutrophil cytoplasm antibody (ANCA)-associated vasculitis. Orbital involvement in WG is a rare but potentially devastating disorder leading to visual loss and deformity. Most clinical characteristics of this disease are non-specific, making clinical diagnosis nearly impossible. We reviewed the features of WG involving the orbit including radiologic, serologic, and histopathologic findings. Advances in treatment are likewise discussed.     

Functional expression of transient receptor potential vanilloid 3 (TRPV3) in corneal epithelial cells: Involvement in thermosensation and wound healing

Transient receptor potential vanilloid 3 (TRPV3), a member of the calcium-permeable thermosensitive TRP (thermoTRP) subfamily of receptors, is an important cutaneous sensor that detects thermal and chemical stimuli. TRPV3 is activated by innocuous warm temperature stimuli (>33° C) and a variety of physiologically active substances. While the corneal epithelium is known to respond to such stimuli, it is unknown whether TRPV3 is involved in this phenomenon. We show here that TRPV3 mRNA and protein are abundantly expressed in the epithelial cells of human and mouse cornea. Carvacrol, an agonist of TRPV3, elevated cytosolic Ca²⁺ concentration in both primary mouse corneal epithelial cells and cultured human corneal epithelial cells (HCE-T cells). The response to carvacrol was inhibited by ruthenium red, a TRPV channel antagonist. Moreover, repetitive agonist stimulation sensitized the response with gradually increasing amplitude, suggesting that the TRPV3 in the cornea has similar physiological and pharmacological characteristics to that in skin keratinocytes. Finally, a wound healing assay revealed that appropriate calcium ion influx via activated TRPV3 in corneal epithelial cells accelerated their proliferation. Thus, functional TRPV3 is present in corneal epithelial cells and may play a role not only in thermosensation, but also in the regulation of cell proliferation.     

m6A甲基化转移酶3在糖尿病性白内障发病中的作用机制

目的:探讨N6-甲基腺嘌呤(N6-methyladenosine, m⁶A)甲基化转移酶3(METTL3)在糖尿病性白内障发病中的作用机制。方法:用低糖和高糖培养基培养人晶状体上皮细胞系(SRA01/04)24h后,采用RT-qPCR和Western blot实验检测细胞的上皮-间质转分化(EMT)指标：E-钙黏蛋白(E-Cadherin)、N-钙黏蛋白(N-Cadherin)、紧密连接蛋白1(ZO-1)和α-平滑肌肌动蛋白(α-SMA)的变化情况;transwell和划痕实验检测细胞迁移能力。采用免疫荧光染色检测人晶状体前囊膜组织中METTL3的表达量及定位,m⁶A dot blot实验检测在低糖和高糖培养基中培养24h细胞的m⁶A甲基化水平,RT-qPCR和Western blot实验检测细胞中METTL3的RNA和蛋白表达量。加入METTL3抑制剂的培养基中培养24h的细胞,RT-qPCR和Western blot实验检测EMT指标的变化情况;m⁶A dot blot实验检测细胞m^6...     

Mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K) pathways differently regulate retinal pigment epithelial cell-mediated collagen gel contraction

Retinal pigment epithelial (RPE) cell-mediated extracellular matrix contraction is believed to contribute to developing proliferative vitreoretinopathy. It has been shown that platelet-derived growth factor (PDGF) and its intracellular signaling pathway, including mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 kinase (PI3K), are mainly involved in this process. The aim of this study is to investigate how these downstream signaling pathways are related to RPE-mediated collagen gel contraction. We performed the gel contraction assay to evaluate the effect of PDGF in cultured ARPE-19 cells under the presence or absence of PD98059, MAPK inhibitor or wortmannin, PI3K inhibitor. Experiments treated with neutralizing antibody for various subtypes of integrin were also performed and the effect on PDGF-induced gel contraction was investigated. Expression changes of integrin alpha1, alpha2 and beta1 after PDGF stimulation was evaluated using quantitative real-time PCR and flow cytometry. The results showed that PDGF up-regulated ARPE-19 cell-mediated gel contractile activity. PDGF-induced collagen gel contraction was attenuated under presence of PD98059, wortmannin, or neutralizing antibody for integrin alpha1, alpha2, or beta1, all of which are critical subset for binding with type I collagen. The expression of integrin alpha1 and alpha2 was increased after PDGF stimulation in both real-time PCR and flow cytometry, however beta1 expression was not increased. PD98059 significantly attenuated integrin alpha1 and alpha2 expressions. However, wortmannin did not have the same effect. In conclusion, PDGF promotes ARPE-19 cell-mediated gel contraction via both MAPK and PI3K. This was probably due to an increased expression of integrin alpha1 and alpha2, which is mediated by MAPK, but not by PI3K. PI3K may regulate collagen gel contraction by another mechanism other than the up-regulation of integrin expression.     

Depletion of optineurin in RGC-5 cells derived from retinal neurons causes apoptosis and reduces the secretion of neurotrophins

Optineurin is a Golgi complex-associated ubiquitous protein with high expression levels in retinal ganglion cells (RGCs). Mutations in optineurin have been observed in rare hereditary cases of primary open-angle glaucoma and in amyotrophic lateral sclerosis. We explored the possibility that optineurin deficiency will compromise neuronal exocytosis leading to a diminished secretion of neurotrophic factors that are critically required for neuronal survival. To this end, we used RNA interference to induce depletion of optineurin in RGC-5 cells derived from retinal neurons. SiRNA specific for optineurin was transiently transfected. Moreover, a stable cell line with constitutive optineurin deficiency (RGC-5 pSilencer OPTN) was generated. In addition, we investigated the subcellular localization of optineurin in primary RGCs in retinal cell cultures isolated from eyes of mature mice. In RGC-5 cells, optineurin localized to the periphery of the Golgi complex and was observed in vesicular structures throughout the cytoplasm and close to the plasma membrane. A comparable Golgi-associated localization of optineurin was observed in cultured primary RGCs that were identified by TUJ1 labeling. Optineurin deficiency caused a marked increase in the number of RGC-5 cells with fragmented Golgi complex. RGC-5 pSilencer OPTN with stable optineurin deficiency showed a pronounced increase in the number of cells undergoing apoptotic cell death. Furthermore, the amounts of secreted neurotrophin-3 (NT-3) and ciliary neurotrophic factor were significantly lower in culture medium of RGC-5 pSilencer OPTN cells when compared to controls. Adding exogenous NT-3 to the culture medium to achieve amounts seen in control cultures completely prevented the increase in apoptotic cell death. We propose that lack of neurotrophic support due to impaired secretion of neurotrophic proteins is a critical factor that causes or contributes to RGC or motor neuron death in patients with mutated optineurin.     

NLRP3炎症小体信号通路在视网膜疾病发生发展中的作用

核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体是由多种蛋白组成的炎症复合物,其主要作用是参与炎症反应。当上述小体激活后可进一步激活Caspase-1,从而诱导一系列炎性因子激活及细胞焦亡。炎性小体的过度活化会引起炎性因子的过量表达,并持续发挥效应,触发免疫失调及炎性连锁反应,造成严重的损害。研究证实糖尿病视网膜病变(DR)、视网膜缺血-再灌注损伤(RIRI)、增生性玻璃体视网膜病变(PVR)等视网膜疾病与免疫失调与炎性反应密切相关,是引起视网膜疾病进展的重要因素。文章就NLRP3炎症小体信号通路及其在视网膜疾病中的功能作一概述,为该病的发病机制及防治提供新思路。