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转化生长因子-β_2对牛眼小梁细胞外基质合成的影响

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中华眼科杂志 2002年第7期38卷 429-432页

作者：曹阳魏厚仁张缨笪邦红卢银平华中科技大学同济医学院附属协和医院眼科武汉430022

目的　观察人眼房水中转化生长因子 β2 (transforminggrowthfactor β2 ,TGF β2 )对体外培养牛眼小梁细胞外基质合成的影响。方法　将体外培养的第 3～ 5代牛眼小梁细胞分为对照组和实验组 ,实验各组分别经 0 32× 10 3 ng/L、1 0 0... 详细信息

目的　观察人眼房水中转化生长因子 β2 (transforminggrowthfactor β2 ,TGF β2 )对体外培养牛眼小梁细胞外基质合成的影响。方法　将体外培养的第 3～ 5代牛眼小梁细胞分为对照组和实验组 ,实验各组分别经 0 32× 10 3 ng/L、1 0 0× 10 3 ng/L、3 2 0× 10 3 ng/LTGF β2 作用 4 8h后 ,分别应用3 H 脯氨酸掺入及液体闪烁测量技术、酶联免疫吸附法和放射免疫技术检测小梁细胞胶原 (collagen ,CA)、纤维连接蛋白 (fibronectin ,FN)及透明质酸 (hyaluronicacid ,HA)的合成。结果　与对照组相比 ,不同浓度的TGF β2 均可促进牛眼小梁细胞CA的合成。 0 32× 10 3 ng/L、1 0 0× 10 3 ng/LTGF β2 组与对照组比较差异有显著意义 (q′=3 85、4 38,P <0 0 5 ) ,3 2 0× 10 3 ng/LTGF β2 组与对照组比较差异有非常显著意义 (q′ =11 4 0 ,P <0 0 1) ,3 H 脯氨酸掺入量呈浓度依赖性增高 ;1 0 0× 10 3 ng/L、3 2 0× 10 3ng/LTGF β2 组促进牛眼小梁细胞合成FN ,与对照组比较差异有显著意义 (q′=3 6 3,P <0 0 5 )和非常显著意义 (q′ =2 0 10 ,P <0 0 1) ;1 0 0× 10 3 ng/L、3 2 0× 10 3 ng/LTGF β2 组均抑制牛眼小梁细胞合成HA ,与对照组比较差异有非常显著意义 (q′=15 15、16 92 ,P <0 0

关键词：转化生长因子-β2 牛眼小梁细胞小梁网细胞外基质原发性开角型青光眼

Tranilast对人眼小梁细胞胶原合成的抑制

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眼科研究 2003年第4期21卷 353-354页

作者：曹阳魏厚仁笪邦红华中科技大学同济医学院附属协和医院眼科武汉430022

目的观察tranilast对体外培养人眼小梁细胞胶原合成的影响.方法体外培养第3～5代牛眼小梁细胞经0 μg/ml(对照组)、12.5、25和50 μg/ml tranilast处理后,采用3H-脯氨酸掺入及液体闪烁测量技术观察tranilast对体外培养人眼小梁细胞胶原... 详细信息

目的观察tranilast对体外培养人眼小梁细胞胶原合成的影响.方法体外培养第3～5代牛眼小梁细胞经0 μg/ml(对照组)、12.5、25和50 μg/ml tranilast处理后,采用3H-脯氨酸掺入及液体闪烁测量技术观察tranilast对体外培养人眼小梁细胞胶原合成的影响.结果 12.5、25和50 μg/ml tranilast处理组3H-脯氨酸掺入率分别为(187±21)%,(127±18)%,(66±12)%,与对照组的(317±48)%比较有极显著差异;同时,3H-脯氨酸掺入率随tranilast质量浓度增加而减少,呈剂量依赖性.结论 tranilast明显抑制小梁细胞胶原的合成.利用tranilast防治原发性开角型青光眼的可能性值得进一步研究.

关键词： Tranilast 人眼小梁细胞胶原合成抑制原发性开角型青光眼

TGF-β2对体外培养牛晶状体上皮细胞CTGF蛋白及mRNA表达的影响

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国际眼科杂志 2008年第4期8卷 701-704页

作者：陈博曹阳笪邦红胡义珍华中科技大学同济医学院附属协和医院眼科中国湖北省武汉市430022 华中科技大学同济医学院附属协和医院急诊科中国湖北省武汉市430022

目的：观察转化生长因子-β2（transforming growth factor-β2，TGF-β2）对体外培养牛眼晶状体上皮细胞（lens epithelial cell，LEC）结缔组织生长因子（connective tissue growth factor，CTGF）mRNA和蛋白表达的影响。方法：将体... 详细信息

目的：观察转化生长因子-β2（transforming growth factor-β2，TGF-β2）对体外培养牛眼晶状体上皮细胞（lens epithelial cell，LEC）结缔组织生长因子（connective tissue growth factor，CTGF）mRNA和蛋白表达的影响。方法：将体外培养2-3代牛LEC分为对照组和实验组，实验组分别经1，5，10μg/LTGF-β2处理24h后，采用半定量RT-PCR和Westernblot技术检测CTGF mRNA以及蛋白表达的变化。结果：与对照组相比，1μg/LTGF-β2组对牛LECCTGF/β-actin吸光度比值和CTGF/β-actin灰度比值的表达无明显影响（P〉0.05），5，10μg/LTGF-β2组可以明显上调RT-PCR和Western blot中CTGF/β-actin比值（P〈0.01）。结论：TGF-β2可促进体外培养牛LEC表达CTGF蛋白及mRNA。

关键词：转化生长因子晶状体上皮细胞结缔组织生长因子白内障

Tranilast对TGF-β_2抑制人眼小梁细胞生长作用的影响

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眼视光学杂志 2002年第2期4卷 92-94页

作者：曹阳魏厚仁笪邦红华中科技大学同济医学院附属协和医院眼科湖北武汉430022

目的 :观察tranilast对转化生长因子 β2 (transform inggrowthfactor β2 ,TGF β2 )抑制体外培养人眼小梁细胞生长作用的影响。方法 :采用MTT法观察 0 μg/ml、12 .5 μg/ml、2 5μg/ml和 5 0 μg/mltranilast对 3.2ng/mlTGF β2 抑... 详细信息

目的 :观察tranilast对转化生长因子 β2 (transform inggrowthfactor β2 ,TGF β2 )抑制体外培养人眼小梁细胞生长作用的影响。方法 :采用MTT法观察 0 μg/ml、12 .5 μg/ml、2 5μg/ml和 5 0 μg/mltranilast对 3.2ng/mlTGF β2 抑制体外培养人眼小梁细胞生长作用的影响。结果 :2 5 μg/ml和 5 0 μg/ml收稿日期 :2 0 0 2 -0 3 -2 7;修回日期 :2 0 0 2 -0 4-15基金项目 :国家自然科学基金资助项目 (3 8970 75 8)。作者简介 :曹阳 (1972 -) ,男 ,湖北武汉人 ,医学博士 ,主治医师 ,研究方向 :青光眼。通信作者 :曹阳 (E -mail:ytsao @sohu .com)。tranilast处理组人眼小梁细胞的光密度A值分别为 (1.136±0 .135 )和 (1.2 4 6± 0 .15 2 ) ,与对照组的 (0 .896± 0 .190 )比较 ,差异分别有显著性 (q =3.2 3,P <0 .0 5 )和非常显著性 (q =4 .70 ,P <0 .0 1)。结论 :Tranilast可明显拮抗TGF β2 对体外培养人眼小梁细胞抑制生长的作用。利用Tranilast在发病学环节防治原发性开角型青光眼的可能性。

关键词： Tranilast TGF-β2 小梁细胞原发性开角型青光眼细胞培养

转化生长因子β_2诱导人小梁细胞凋亡的体外研究

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眼科新进展 2004年第1期24卷 12-14页

作者：笪邦红曹阳魏厚仁华中科技大学同济医学院附属协和医院急诊科湖北武汉430022 华中科技大学同济医学院附属协和医院眼科湖北武汉430022

目的　观察转化生长因子 β2 (transforminggrowthfactor β2 ,TGF β2 )是否能诱导体外培养人眼小梁细胞 (humantrabecularmeshworkcells,HTMC)发生凋亡。方法　将体外培养的第 3～ 5代人眼小梁细胞分为对照组和实验组 ,实验各组分别经... 详细信息

目的　观察转化生长因子 β2 (transforminggrowthfactor β2 ,TGF β2 )是否能诱导体外培养人眼小梁细胞 (humantrabecularmeshworkcells,HTMC)发生凋亡。方法　将体外培养的第 3～ 5代人眼小梁细胞分为对照组和实验组 ,实验各组分别经 0 .32、1.0 0、3.2μg·L-1TGF β2 作用 4 8h后 ,分别用透射电镜、TUNEL法和流式细胞术检测细胞凋亡的诱导情况。结果　透射电镜观察发现凋亡细胞典型的形态学变化———核染色质凝集边集 ,凋亡小体形成等。TUNEL法检出发生DNA片段化的HTMC。经流式细胞术定量分析 ,不同浓度TGF β2 处理的HTMC凋亡细胞百分数分别为 ( 2 .79± 0 .4 4 ) %、( 4 .4 3±1.17) %、( 9.6 0± 2 .0 5 ) % ,其中 1、3.2 μg·L-1TGF β2 处理组与对照组 ( 1.4 1± 0 .34) %相比分别有显著差异和极显著差异。结论　TGF β2 能诱导体外培养HTMC发生凋亡 ,推测其可能参与了原发性开角型青光眼患者和正常人衰老过程中HTMC数目的减少。

关键词：转化生长因子小梁网细胞培养凋亡

Transforming Growth Factor-β_2 Gene Cloning and Protein Expression in Human Trabecular Meshwork Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2003年第1期23卷 85-87页

作者：曹阳魏厚仁笪邦红李忠玉 DepartmentofOphthalmology XieheHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnology DepartmentofEmergency XieheHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnology DepartmentofAnatomy TongjiMedicalCollegeHuazhongUniversityofScienceandTechnology

Whether cultured human trabecular meshwork cells express transforming growth factor-β 2 (TGF-β 2) messenger RNA (mRNA) and protein was investigated. Total RNA of 10 6 cultured human trabecular meshwork cells was extracted with TRIZOL reagent, reverse transcriptase-polymerase chain reaction (RT-PCR) were used for detection of TGF-β 2 messenger RNA, and the PCR product was verified by sequencing. Immunohistochemical staining was used to detect TGF-β 2 protein. The results showed that a single RT-PCR amplified product was obtained, and the sequence was homologous to the known sequence. TGF-β 2 immunostaining was positive. It was concluded that trabecular meshwork cells could produce TGF-β 2 and contribute to the presence of TGF-β 2 in trabecular meshwork microenvironment as well as aqueous humor. Trabecular meshwork cells were affected by TGF-β 2 not only through paracrine, but also autocrine action. Whether abnormal changes in TGF-β 2 production contribute to the pathogenesis of primary open-angle glaucoma is worth further investigation.

关键词： human trabecular meshwork cultured cells transforming growth factor reverse transcriptase-polymerase chain reaction immunohistochemistry

Antagonistic Effects of Tranilast on Proliferation and Collagen Synthesis Induced by TGF-β_2 in Cultured Human Trabecular Meshwork Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2004年第5期24卷 490-492,496页

作者：笪邦红曹阳魏厚仁陈志新水迎波李忠玉 Department of Emergency Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China Department of Ophthalmology Union Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan 430022 China Department of Ophthalmology Hanchuan TCM Hospital Hanchuan 432300 China Department of Ophthalmology and Visual Sciences Washington University School of Medicine St. Louis 63110 United States Department of Anatomy Tongji Medical College of Huazhong University of Science and Technology Wuhan 430030 China

Whether tranilast had antagonistic effect on proliferation inhibition and collagen synthesis promotion induced by TGF-β 2 in cultured human trabecular meshwork cells was investigated. Suspension of 1×104 cultured human trabecular meshwork cells of 3—5 passage was distributed in each well of a 96-well disk and divided into control group and experimental group. After 24 h, 0 μg/ml (control), 12.5 μg/ml, 25 μg/ml, 50 μg/ml tranilast with 3.2 ng/ml TGF-β 2 were added into the incubation medium. Another 24 h later, proliferation and collagen synthesis in cultured human trabecular meshwork cells were examined respectively by using tetrazolium-based semiautomated colormetric (MTT) assay and 3H-proline incorporation with liquid scintillation technique. The results showed absorbance (A) values of the experimental groups were 0.9036±0.3017, 1.1361± 0.1352, 1.2457±0.1524 according to the different concentrations of tranilast, and 0.8956± 0.1903 of the control group. In comparison with the control group, 25 μg/ml (q'=3.23, P< 0.05), 50 μg/ml (q'=4.70, P<0.01) tranilast significantly antagonized the decrease of the A values induced by TGF-β 2 in the cultured human trabecular meshwork cells. In comparison with the control group [817.37±124.21 cpm/104 cells], 12.5 μg/ml (620.33±80.46 cpm/104 cells, q'= 4.26, P< 0.05), 25 μg/ml (594.58±88.13 cpm/104 cells, q'=4.81, P<0.01), 50 μg/ml (418.64±67.90 cpm/104 cells, q'=8.62, P<0.01) tranilast significantly inhibited the incorporation of 3H-proline into the cultured human trabecular meshwork cells promoted by TGF-β 2 in a dose-dependent manner. It was concluded that tranilast had the antagonistic effect on the proliferation inhibition and collagen synthesis promotion induced by TGF-β 2 in the cultured human trabecular meshwork cells.

关键词： transforming growth factor human trabecular meshwork cultured cell proliferation collagen

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妊娠期肝内胆汁淤积症的诊断与治疗

中国实用妇科与产科杂志 2001年第6期17卷 328-330页

作者：王泽华笪邦红华中科技大学同济医学院附属协和医院妇产科武汉430022

Effect of Transforming Growth Factor-β_2 on Phagocytosis in Cultured Bovine Trabecular Meshwork Cells

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2001年第4期21卷 318-320页

作者：曹阳魏厚仁笪邦红黄毅 Department of Ophthalmology Xiehe Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan

The effect of transforming growth factor β 2 (TGF β 2) on phagocytosis in bovine trabecular meshwork cells in vitro was investigated. After the cultured bovine trabecular meshwork cells were treated with 0 ng/ml, 0.32 ng/ml, 1 ng/ml, 3.2 ng/ml TGF β 2 for 24 h, latex beads were added into the incubation medium, and the numbers of the latex beads in 20 adjacent cells were counted under a microscope 24 h later, after treatment with Wright's stain. Our results showed that the average numbers of the latex beads in the trabecular meshwork cells treated with TGF β 2 of different concentrations were 53.1±1.7 beads/cell, 56.4±2.9 beads/cell and 77.9±6.5 beads/cell respectinvely, in comparison with 45.5±3.3 beads/cell of the control group. TGF β 2 significantly increased the number of the latex beads phagocytosed by cultured bovine trabecular meshwork cells in a dose dependent manner. TGF β 2 could promote the phagocytosis of bovine trabecular meshwork cells in vitro . It may be involved in the cellularity decrease of the trabecular meshwork in the patients of primary open angle glaucoma through promoting the phagocytosis of trabecular meshwork cells.

关键词： transforming growth factor trabecular meshwork cultured cells phagocytosis