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色素上皮衍生因子对晶状体上皮细胞生长的促进作用及其机制 被引量:3
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作者 刘恬 刘奕志 +1 位作者 钟惟德 项道满 《中华实验眼科杂志》 CAS CSCD 北大核心 2015年第4期342-346,共5页
背景 晶状体上皮细胞(LECs)是研究晶状体生理病理的基础,色素上皮衍生因子(PEDF)是存在于房水和晶状体中的多效能因子,但PEDF对LECs的生物学效应及其机制尚需深入探讨.目的 探讨PEDF在体内及体外对人LECs生长的调节作用及其机制.方... 背景 晶状体上皮细胞(LECs)是研究晶状体生理病理的基础,色素上皮衍生因子(PEDF)是存在于房水和晶状体中的多效能因子,但PEDF对LECs的生物学效应及其机制尚需深入探讨.目的 探讨PEDF在体内及体外对人LECs生长的调节作用及其机制.方法 在体内研究部分,纳入年龄相关性白内障患者181例181眼,在白内障手术过程中取患眼晶状体中央区5.0~5.5 mm直径的前囊膜标本,检测LECs密度并依此选择其中60份分为低密度组和高密度组各30例.采用逆转录PCR(RT-PCR)法检测并比较2个组LECs中PEDF mRNA相对表达量的差异.在体外研究部分,PEDF培养组在培养液中加入50 ng/ml PEDF培养人LECs株(HLE-B3)72 h,正常培养的细胞作为对照组,采用流式细胞仪通过AnnexinV-FITC/7-AAD双染法检测并比较各组细胞周期的细胞比例和细胞凋亡率,采用实时荧光定量PCR(RT-qPCR)法检测细胞中VEGF mRNA的相对表达量.结果 低密度组晶状体中央区前囊下LECs密度为(3 672±326)/mm2,PEDFmRNA相对表达量为0.43±0.05,明显低于高密度组的(4 857±350)/mm2和0.55±0.04,差异均有统计学意义(t=4.16,P<0.05;t=3.82,P<0.05).PEDF培养组G2 +S期细胞比例为(54.05±4.98)%,明显高于对照组的(28.54±4.27)%,差异有统计学意义(t=-6.32,P<0.01);PEDF培养组细胞凋亡率为(2.08±0.48)%,明显低于对照组的(13.50±0.72)%,差异有统计学意义(t=7.90,P<0.01).PEDF培养组细胞内VEGF mRNA相对表达量为6.1±2.2,明显低于对照组的27.4±4.8,差异有统计学意义(t=5.48,P<0.01).结论 人眼内PEDF可通过抗凋亡及抑制VEGF表达等机制发挥促LECs增生的作用,LECs中PEDF的变化可能与晶状体的老化及白内障的发生和发展有关. 展开更多
关键词 晶状体/细胞学 上皮细胞/细胞学 眼蛋白/代谢 细胞培养 血管内皮生长因子 色素上皮衍生因子
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Wnt3a诱导人晶状体上皮细胞细胞外基质合成和胶原凝胶收缩的机制研究 被引量:2
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作者 包秀丽 刘婷婷 +1 位作者 王艳艳 李琳 《中华实验眼科杂志》 CAS CSCD 北大核心 2016年第7期597-601,共5页
背景晶状体上皮细胞(LECs)的细胞外基质(ECM)的产生、异常沉积和收缩是PCO发生的重要病理机制。Wnt3a蛋白参与机体组织上皮细胞ECM的产生和纤维化过程,但Wnt3a对晶状体组织中上皮间质转化(EMT)相伴行的ECM的产生和收缩的影响尚... 背景晶状体上皮细胞(LECs)的细胞外基质(ECM)的产生、异常沉积和收缩是PCO发生的重要病理机制。Wnt3a蛋白参与机体组织上皮细胞ECM的产生和纤维化过程,但Wnt3a对晶状体组织中上皮间质转化(EMT)相伴行的ECM的产生和收缩的影响尚不清楚。目的研究Wnt3a对体外培养的人LECsECM合成的影响,并探讨其介导胶原凝胶收缩的相关分子机制。方法用脂质体介导转染技术将Writ3acDNA表达载体瞬时转染人LECs系SRAOI/04细胞作为Wnt3a转染组,对照组转染pcDNA3-HA表达载体。细胞转染后48h,采用Westernblot法测定各组细胞中Wnt3a、ECM主要成分I型胶原纤维(Col-I),Ⅳ型胶原纤维(C01.IV)和整合素B1(integrinp1)的表达;采用免疫荧光法检测α-SMA、细胞纤维状肌动蛋白(F-actin)在胞中的表达和分布。将SRAOI/04细胞与I型胶原凝胶混合,观察胶原收缩情况。结果Wnt3a转染48h,SRAOI/04细胞内Writ3a蛋白相对表达量(相对灰度值)明显升高,对照组细胞和Wnt3a转染组细胞中Wnt3a蛋白表达量分别为0.290±0.066和0.703±0.105,差异有统计学意义(t=5.782,P〈0.01)。Westernblot法检测显示,Wnt3a转染组SRAOI/04细胞中的Col-I和integrinp1蛋白的相对表达量分别为0.697±0.021和0.875±0.055,明显高于对照组的0.370±0.020和0.580±0.030,差异均有统计学意义(t=19.600、8.156,均P〈0.01),Wnt3a转染组Col—IV蛋白相对表达量为0.430±O.020,高于对照组的0.383±0.031,但差异无统计学意义(t=2.514,P〉0.05)。免疫荧光检测显示,对照组细胞中F-actin和α-SMA蛋白主要表达于细胞膜,而Wnt3a转染组细胞中F-aetin和α-SMA蛋白表达于细胞膜、细胞质和细胞核周围,阳性染色程度较对照组明显增强。Col-I凝胶收缩试验结果显示,细胞与Col-I胶原凝胶混合后24h凝胶收缩面积比率明显低于混合后8h(64.1%±2.3%与98.9%±1.0%),Wnt3a转染组凝胶收缩面积比率明显低于对照组(64.1%±2.3%与93.9%±3.1%),差异均有统计学意义(均P〈0.01)。结论Wnt3a过表达可诱导SRA01/04细胞ECM合成和细胞骨架重建,促进细胞收缩。 展开更多
关键词 上皮间质转化 晶状体上皮细胞 晶状体/细胞学 细胞外基质 后囊膜混浊 Wnt信号通路 转染
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Bloom Forming Species of Phytoplankton in Two Coastal Waters in the Southeast Coast of India 被引量:1
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作者 K. Thillai Rajasekar M. Rajkumar +2 位作者 SUN Jun V. Ashok Prabu E Perumal 《Journal of Ocean University of China》 SCIE CAS 2010年第3期265-272,共8页
The results of an investigation carried out during June 2005 to May 2007 on bloom-forming phytoplankton species composition and abundance in the Parangipettai and Coleroon coastal waters (Southeast coast of India) a... The results of an investigation carried out during June 2005 to May 2007 on bloom-forming phytoplankton species composition and abundance in the Parangipettai and Coleroon coastal waters (Southeast coast of India) are reported. Air and surface water temperatures (℃) varied from 25.1 to 30.1 and 24.5 to 28.5, respectively, in the former waters and from 25.5 to 31.2 and 25.0 to 29.3 in the latter waters. The respective salinities varied from 6.0 to 28.5 and 5.0 to 33.1 and the respective pH ranged between 7.0 and 8.3 and 7.2 and 8.3. Correspondingly, the dissolved oxygen content varied from 3.1 to 7.5 and 3.1 to 7.9 mgL1 while the light extinction coefficient (LEC) values ranged between 3.1 and 10.1 and 1.8 and 11.0. The content ranges of inorganic nutrients, i.e., nitrate, nitrite, phosphate and silicate (μmolL^-1), in the Parangipettai and Coleroon coastal waters were: 6.5- 27.0; 1.0- 8.9; 0.1-3.0 and 15.0 -140 and 10.1-23.4; 1.2-8.9; 0.2-3.1 and 55-125 respectively. The chlorophyll α contents in both waters ranged from 2.0-7.5 μgL^-1. Presently, 124 phytoplankton species representing different classes were recorded in the Coleroon coast, viz, Bacillariophyceae (77); Dinophyceae (19); Cyanophyceae (15); Chlorophyceae (10) and Chrysophyceae (3), whereas 117 phytoplankton species were recorded in the Parangipettai coast, viz, Bacillariophyceae (66); Dinophyceae (22); Cyanophyceae (19); Chlorophyceae (7) and Chrysophyceae (3). The phytoplankton cell abundance in the Parangipettai and Coleroon coastal waters varied from 290 to 111662 and 140 to 132 757 cells L^-1, respectively, with peak diversity (3.38 and 3.52 bits indl.) recorded in summer. The maximum abundance occurred in summer coinciding with the stable hydrographical conditions. The seasonal distribution and abundance of phytoplankton are discussed in relation to hydrographical parameters. Totally 31 and 24 species of phytoplankton were found to be bloom-forming in the Parangipettai and Coleroon coastal waters, respectively. Presently Trichodesmium bloom was also observed, which appeared at the 10 fathom level of the coastal water and quickly spread to the marine zone of the Vellar estuary and near the mouth region of the mangrove waters. The Parangipettai and Coleroon coastal waters are subject to long term fluctuations in physico-chemical conditions depending upon the seasonal tidal range and freshwater influx, resulting in a continuous exchange of organic, inorganic, plant and animal matters. 展开更多
关键词 Bloom-forming phytoplankton SPECIES ABUNDANCE Parangipettai and Coleroon coastal waters
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