中华眼科杂志
中華眼科雜誌
중화안과잡지
Chinese Journal of Ophthalmology
2012年
7期
619-624
,共6页
富含半胱氨酸蛋白质61%脉络膜%视网膜%内皮细胞%视网膜新生血管化%细胞增殖%细胞运动
富含半胱氨痠蛋白質61%脈絡膜%視網膜%內皮細胞%視網膜新生血管化%細胞增殖%細胞運動
부함반광안산단백질61%맥락막%시망막%내피세포%시망막신생혈관화%세포증식%세포운동
Cysteine-rich protein 61%Choroid%Retinal%Endothelial cells%Retinal neovascularization%Cell proliferation%Cell movement
目的 研究富含半胱氨酸蛋白61(CYR61)对脉络膜视网膜内皮细胞增殖、迁移及形成管状结构的影响.方法 实验研究.采用噻唑蓝比色法,观察CYR61对体外培养的非洲绿猴脉络膜视网膜内皮细胞系(RF-6A)增殖能力的影响；应用Transwell小室实验,观察在CYR61作用下RF-6A细胞迁移能力的变化；采用Matrigel基质胶实验,观察CYR61对RF-6A细胞形成管状结构的影响.不同浓度CYR61和不同于预时间的吸光度(A)值、各组迁移的细胞数和管状结构数比较,采用单因素方差分析.结果 应用不同浓度的CYR61(O、5、10、50、100及500 μg/L)干预RF-6A细胞72 h,随着CYR61浓度增加,MTT比色法显示A490nm值升高,分别为0.511、0.522、0.532、0.597、0.765、0.818,差异有统计学意义(F=318.828,P ＜0.05)；400 μg/L的CYR61干预RF-6A细胞不同时间(O、12、24、48及72 h),随着时间的延长,A490nm值升高,分别为0.533、0.598、0.643、0.695及0.756,差异有统计学意义(F=42.910,P＜0.05).Transwell小室实验中,不同浓度的CYR61(40、200、400 μg/L的CYR61及400μg/L的CYR61 +25 mg/L的抗CYR61抗体)、80 μg/L的血管内皮生长因子(VEGF)及阴性对照组每一高倍视野的细胞数分别为(66.83±3.87)、(77.83±4.26)、(96.83±3.49)、(70.67±3.83)、(98.33±3.14)及(62.00±7.62)个,随着CYR61浓度增加,迁移的RF-6A细胞数增加(F=46.987,P＜0.05),400 μ,g/L的CYR61促进细胞迁移的能力与80 μg/L的VEGF相当,而加入抗CYR61抗体则可以起到显著的抑制作用.Matrigel基质胶实验中,不同浓度的CYR61( 40、200、400 μg/L的CYR61及400 μg/L的CYR61 +25 mg/L的抗CYR61抗体)、80 μg/L的VEGF及阴性对照组每一高倍视野的管状结构数分别为(34.33±2.50)、(60.67±3.72)、(88.17±2.93)、(51.17±2.14)、(90.83±3.49)及(31.83±3.31)个,随着CYR61浓度增加,RF-6A细胞形成的管状结构数增多(F=355.224,P ＜0.05),400 μg/L的CYR61与80 μg/L的VEGF作用相当,而加入抗CYR61抗体则可以起到显著的抑制作用.结论 CYR61在体外可以促进脉络膜视网膜内皮细胞增殖、迁移及形成管状结构,CYR61可能在视网膜新生血管的形成过程中起一定作用.
目的 研究富含半胱氨痠蛋白61(CYR61)對脈絡膜視網膜內皮細胞增殖、遷移及形成管狀結構的影響.方法 實驗研究.採用噻唑藍比色法,觀察CYR61對體外培養的非洲綠猴脈絡膜視網膜內皮細胞繫(RF-6A)增殖能力的影響；應用Transwell小室實驗,觀察在CYR61作用下RF-6A細胞遷移能力的變化；採用Matrigel基質膠實驗,觀察CYR61對RF-6A細胞形成管狀結構的影響.不同濃度CYR61和不同于預時間的吸光度(A)值、各組遷移的細胞數和管狀結構數比較,採用單因素方差分析.結果 應用不同濃度的CYR61(O、5、10、50、100及500 μg/L)榦預RF-6A細胞72 h,隨著CYR61濃度增加,MTT比色法顯示A490nm值升高,分彆為0.511、0.522、0.532、0.597、0.765、0.818,差異有統計學意義(F=318.828,P ＜0.05)；400 μg/L的CYR61榦預RF-6A細胞不同時間(O、12、24、48及72 h),隨著時間的延長,A490nm值升高,分彆為0.533、0.598、0.643、0.695及0.756,差異有統計學意義(F=42.910,P＜0.05).Transwell小室實驗中,不同濃度的CYR61(40、200、400 μg/L的CYR61及400μg/L的CYR61 +25 mg/L的抗CYR61抗體)、80 μg/L的血管內皮生長因子(VEGF)及陰性對照組每一高倍視野的細胞數分彆為(66.83±3.87)、(77.83±4.26)、(96.83±3.49)、(70.67±3.83)、(98.33±3.14)及(62.00±7.62)箇,隨著CYR61濃度增加,遷移的RF-6A細胞數增加(F=46.987,P＜0.05),400 μ,g/L的CYR61促進細胞遷移的能力與80 μg/L的VEGF相噹,而加入抗CYR61抗體則可以起到顯著的抑製作用.Matrigel基質膠實驗中,不同濃度的CYR61( 40、200、400 μg/L的CYR61及400 μg/L的CYR61 +25 mg/L的抗CYR61抗體)、80 μg/L的VEGF及陰性對照組每一高倍視野的管狀結構數分彆為(34.33±2.50)、(60.67±3.72)、(88.17±2.93)、(51.17±2.14)、(90.83±3.49)及(31.83±3.31)箇,隨著CYR61濃度增加,RF-6A細胞形成的管狀結構數增多(F=355.224,P ＜0.05),400 μg/L的CYR61與80 μg/L的VEGF作用相噹,而加入抗CYR61抗體則可以起到顯著的抑製作用.結論 CYR61在體外可以促進脈絡膜視網膜內皮細胞增殖、遷移及形成管狀結構,CYR61可能在視網膜新生血管的形成過程中起一定作用.
목적 연구부함반광안산단백61(CYR61)대맥락막시망막내피세포증식、천이급형성관상결구적영향.방법 실험연구.채용새서람비색법,관찰CYR61대체외배양적비주록후맥락막시망막내피세포계(RF-6A)증식능력적영향；응용Transwell소실실험,관찰재CYR61작용하RF-6A세포천이능력적변화；채용Matrigel기질효실험,관찰CYR61대RF-6A세포형성관상결구적영향.불동농도CYR61화불동우예시간적흡광도(A)치、각조천이적세포수화관상결구수비교,채용단인소방차분석.결과 응용불동농도적CYR61(O、5、10、50、100급500 μg/L)간예RF-6A세포72 h,수착CYR61농도증가,MTT비색법현시A490nm치승고,분별위0.511、0.522、0.532、0.597、0.765、0.818,차이유통계학의의(F=318.828,P ＜0.05)；400 μg/L적CYR61간예RF-6A세포불동시간(O、12、24、48급72 h),수착시간적연장,A490nm치승고,분별위0.533、0.598、0.643、0.695급0.756,차이유통계학의의(F=42.910,P＜0.05).Transwell소실실험중,불동농도적CYR61(40、200、400 μg/L적CYR61급400μg/L적CYR61 +25 mg/L적항CYR61항체)、80 μg/L적혈관내피생장인자(VEGF)급음성대조조매일고배시야적세포수분별위(66.83±3.87)、(77.83±4.26)、(96.83±3.49)、(70.67±3.83)、(98.33±3.14)급(62.00±7.62)개,수착CYR61농도증가,천이적RF-6A세포수증가(F=46.987,P＜0.05),400 μ,g/L적CYR61촉진세포천이적능력여80 μg/L적VEGF상당,이가입항CYR61항체칙가이기도현저적억제작용.Matrigel기질효실험중,불동농도적CYR61( 40、200、400 μg/L적CYR61급400 μg/L적CYR61 +25 mg/L적항CYR61항체)、80 μg/L적VEGF급음성대조조매일고배시야적관상결구수분별위(34.33±2.50)、(60.67±3.72)、(88.17±2.93)、(51.17±2.14)、(90.83±3.49)급(31.83±3.31)개,수착CYR61농도증가,RF-6A세포형성적관상결구수증다(F=355.224,P ＜0.05),400 μg/L적CYR61여80 μg/L적VEGF작용상당,이가입항CYR61항체칙가이기도현저적억제작용.결론 CYR61재체외가이촉진맥락막시망막내피세포증식、천이급형성관상결구,CYR61가능재시망막신생혈관적형성과정중기일정작용.
Objective To observe the effects of GYR61 (cysteine-rich 61; CCN1 ) on the proliferation,migration and tube formation of choroid-retinal endothelial cells (RF-6A cell line).Methods Experimental study.RF-6A cells were cultured and treated with CYR61 at different concentrations.Effects of CYR61 on cell proliferation,migration and angiogenesis were observed by MTT assay,transwell assay and tube formatiou assay.Results When different concentrations of CYR61 (0,5,10,100 and 500 μg/L)were used to treat RF-6A cells for 72 h,A490 nm value of the MTT assay was changed dose-dependently (0.511,0.522,0.532,0.597,0.765 and 0.818),and the difference between different dosage groups was statistically significant ( F =318.828,P ＜ 0.05 ).When RF-6A cells were treated with 400 μg/L CYR61 for different time periods (0,12,24,48 and 72 h),A490 nm value increased with the extension of treatment time (0.533,0.598,0.643,0.695 and 0.756 ),and the difference was statistically significant (F =42.910,P ＜ 0.05 ).In transwell assay,migrated cells in cells treated with different concentrations of CYR61 (40,200,400 μg/L and 400 μμg/L + 25 mg/Lanti-CYR61 antibody),80 μg/L VEGF,and negative control groups were 66.83 ± 3.87,77.83 ± 4.26,96.83 ± 3.49,70.67 ± 3.83,98.33 ± 3.14 and 62.00 ± 7.62per high-power field,respectively.RF-6A cell migration capacity increased with increased concentration of CYR61 (F =46.987,P ＜ 0.05 ).In tube formation assay,numbers of tube in different concentrations of CYR61 (40,200,400 μg/L,400 μg/L + 25 mg/L anti-CYR61 antibody),80 μg/L VEGF and negative control groups were 34.33 ±2.50,60.67 ±3.72,88.17 ±2.93,51.17 ±2.14,90.83 ±3.49 and 31.83 ± 3.31 per field.RF-6A cell tube formation capacity increased with increased concentration of CYR61 ( F =355.224,P ＜ 0.05 ).There were equal effects between 400 μg/L CYR61 and 80 μg/L VEGF.AntiCYR61 antibody could inhibit cell migration and tube formation promoted by CYR61.Conclusions CYR61 can promote proliferation,migration and tnbe formation of choroid-retinal endothelial cells in vitro.CYR61 are likely to be involved in the pathogenesis of retinal neovascularization.
