中国实用眼科杂志
中國實用眼科雜誌
중국실용안과잡지
CHINESE JOURNAL OF PRACTICAL OPHTHALMOLOGY
2013年
6期
779-783
,共5页
蓝光%视网膜色素上皮细胞%复制衰老%炎症%趋化因子
藍光%視網膜色素上皮細胞%複製衰老%炎癥%趨化因子
람광%시망막색소상피세포%복제쇠로%염증%추화인자
Blue light%Retinal pigment epithelium%Replicative senescence%Inflammation%Chemotactic factor
目的 观察蓝光能否诱导人RPE细胞发生复制衰老及对趋化因子表达的影响,为探讨蓝光及炎症在年龄相关性黄斑变性(AMD)发病机制中的作用提供实验依据.方法 取猝死的健康成年人眼球,采用胰蛋白酶消化法获取RPE细胞,培养后经免疫组化鉴定细胞纯度.采用不同强度(2000±500) lux、(1000±200) lux、(500±100) lux的蓝光,每天光照1h,连续照射4d,诱导RPE细胞复制衰老,并用衰老相关-β半乳糖苷酶(SA-β-Gal)进行检测.用RT-PCR法检测细胞内趋化因子的表达情况.结果 用胰蛋白酶消化法,能够获得RPE细胞,并且体外能稳定增殖.蓝光能够诱导RPE细胞发生复制衰老,随着蓝光光照度的增加,复制衰老阳性细胞数目逐渐增多,细胞内趋化因子IL-8、MCP-1的表达增加,差异具有统计学意义(P＜0.05).结论 蓝光可以诱导人RPE细胞发生复制衰老,复制衰老的RPE细胞内趋化因子IL-8、MCP-1的表达增加.
目的 觀察藍光能否誘導人RPE細胞髮生複製衰老及對趨化因子錶達的影響,為探討藍光及炎癥在年齡相關性黃斑變性(AMD)髮病機製中的作用提供實驗依據.方法 取猝死的健康成年人眼毬,採用胰蛋白酶消化法穫取RPE細胞,培養後經免疫組化鑒定細胞純度.採用不同彊度(2000±500) lux、(1000±200) lux、(500±100) lux的藍光,每天光照1h,連續照射4d,誘導RPE細胞複製衰老,併用衰老相關-β半乳糖苷酶(SA-β-Gal)進行檢測.用RT-PCR法檢測細胞內趨化因子的錶達情況.結果 用胰蛋白酶消化法,能夠穫得RPE細胞,併且體外能穩定增殖.藍光能夠誘導RPE細胞髮生複製衰老,隨著藍光光照度的增加,複製衰老暘性細胞數目逐漸增多,細胞內趨化因子IL-8、MCP-1的錶達增加,差異具有統計學意義(P＜0.05).結論 藍光可以誘導人RPE細胞髮生複製衰老,複製衰老的RPE細胞內趨化因子IL-8、MCP-1的錶達增加.
목적 관찰람광능부유도인RPE세포발생복제쇠로급대추화인자표체적영향,위탐토람광급염증재년령상관성황반변성(AMD)발병궤제중적작용제공실험의거.방법 취졸사적건강성년인안구,채용이단백매소화법획취RPE세포,배양후경면역조화감정세포순도.채용불동강도(2000±500) lux、(1000±200) lux、(500±100) lux적람광,매천광조1h,련속조사4d,유도RPE세포복제쇠로,병용쇠로상관-β반유당감매(SA-β-Gal)진행검측.용RT-PCR법검측세포내추화인자적표체정황.결과 용이단백매소화법,능구획득RPE세포,병차체외능은정증식.람광능구유도RPE세포발생복제쇠로,수착람광광조도적증가,복제쇠로양성세포수목축점증다,세포내추화인자IL-8、MCP-1적표체증가,차이구유통계학의의(P＜0.05).결론 람광가이유도인RPE세포발생복제쇠로,복제쇠로적RPE세포내추화인자IL-8、MCP-1적표체증가.
Objective To observe the effects of blue light on human RPE cells replicative senescence and expression of chemokines in RPE cell,and to discuss the role of blue light and inflammatory reaction in pathogenesis of age-related macular degeneration (AMD).Methods Healthy human eyeball was used for experiments.Primary human RPE culture was harvested with trypsin digestion.The cells were identified with immunohistochemistry.The blue light source of light emitting diode (2000±500)lux,(1000±200)lux,(500±100)lux were used to cause replicative senescence 1 hour a day,4 days of continuous irradiation and detect senescence associated beta-galactosidae (SA-β-Gal) activity.RT-PCR assay was used to detect expressions of chemokines IL-8,MCP-1 in RPE cells.Results This primary culture resulted in cells with well-preserved morphology using trypsin.The next passages were grown well.The number of the replicative senescence cells and the expressions chemokines IL-8,MCP-1 in RPE cells with blue particles increased as the increase of exposure intensity.The difference had statistics significance (P ＜0.05).Conclusions The replicative senescence of human RPE cells is positive correlation with blue light.The blue light increases the expressions of chemokins IL-8,MCP-1 in human RPE cells.
