中华眼底病杂志
中華眼底病雜誌
중화안저병잡지
CHINESE JOURNAL OF OCULAR FUNDUS DISEASES
2010年
6期
544-547
,共4页
曲安奈德/药理学%趋化因子CXCL12
麯安奈德/藥理學%趨化因子CXCL12
곡안내덕/약이학%추화인자CXCL12
Triamcinolone acetonide/pharmacology%Chemokine CXCL12
目的 观察曲安奈德(TA)对人视网膜色素上皮细胞衍生因子(PEDF)表达的影响.方法 体外培养人视网膜色素上皮(RPE)细胞,取第4～6代细胞用于实验.分别以浓度为40、400、4×103、4×104μg/L的TA干预12、24、48 h后,采用蛋白质免疫印迹法(Western blot)检测培养液及细胞浆中PEDF蛋白表达水平.采用20 ng/ml的肿瘤坏死因子-α(TNF-α)预干预RPE细胞24 h后,再加入400μg/L的TA进行干预(TNF-α预处理组);分别选用20 ng/ml的TNF-α和400μg/L的TA单独干预RPE细胞作为单独干预组.分别作用1、6、24 h后,测定3组细胞培养液及细胞浆中PEDF及细胞浆中磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)的蛋白表达水平.结果 各TA浓度组RPE细胞培养液及细胞浆中PEDF蛋白表达均升高,其中400μg/L组PEDF蛋白表达最高(P＜0.05).干预1、6、24 h时,TNF-α预处理组和TA单独干预组PEDF蛋白表达均增高、p-p38MAPK蛋白表达均降低(P＜0.01);TNF-α单独干预组PEDF蛋白表达降低、p-p38MAPK蛋白表达增高(P＜0.01).结论 TA能够上调体外培养的人RPE细胞PEDF蛋白表达,下调p-p38MAPK蛋白表达.
目的 觀察麯安奈德(TA)對人視網膜色素上皮細胞衍生因子(PEDF)錶達的影響.方法 體外培養人視網膜色素上皮(RPE)細胞,取第4～6代細胞用于實驗.分彆以濃度為40、400、4×103、4×104μg/L的TA榦預12、24、48 h後,採用蛋白質免疫印跡法(Western blot)檢測培養液及細胞漿中PEDF蛋白錶達水平.採用20 ng/ml的腫瘤壞死因子-α(TNF-α)預榦預RPE細胞24 h後,再加入400μg/L的TA進行榦預(TNF-α預處理組);分彆選用20 ng/ml的TNF-α和400μg/L的TA單獨榦預RPE細胞作為單獨榦預組.分彆作用1、6、24 h後,測定3組細胞培養液及細胞漿中PEDF及細胞漿中燐痠化p38絲裂原活化蛋白激酶(p-p38MAPK)的蛋白錶達水平.結果 各TA濃度組RPE細胞培養液及細胞漿中PEDF蛋白錶達均升高,其中400μg/L組PEDF蛋白錶達最高(P＜0.05).榦預1、6、24 h時,TNF-α預處理組和TA單獨榦預組PEDF蛋白錶達均增高、p-p38MAPK蛋白錶達均降低(P＜0.01);TNF-α單獨榦預組PEDF蛋白錶達降低、p-p38MAPK蛋白錶達增高(P＜0.01).結論 TA能夠上調體外培養的人RPE細胞PEDF蛋白錶達,下調p-p38MAPK蛋白錶達.
목적 관찰곡안내덕(TA)대인시망막색소상피세포연생인자(PEDF)표체적영향.방법 체외배양인시망막색소상피(RPE)세포,취제4～6대세포용우실험.분별이농도위40、400、4×103、4×104μg/L적TA간예12、24、48 h후,채용단백질면역인적법(Western blot)검측배양액급세포장중PEDF단백표체수평.채용20 ng/ml적종류배사인자-α(TNF-α)예간예RPE세포24 h후,재가입400μg/L적TA진행간예(TNF-α예처리조);분별선용20 ng/ml적TNF-α화400μg/L적TA단독간예RPE세포작위단독간예조.분별작용1、6、24 h후,측정3조세포배양액급세포장중PEDF급세포장중린산화p38사렬원활화단백격매(p-p38MAPK)적단백표체수평.결과 각TA농도조RPE세포배양액급세포장중PEDF단백표체균승고,기중400μg/L조PEDF단백표체최고(P＜0.05).간예1、6、24 h시,TNF-α예처리조화TA단독간예조PEDF단백표체균증고、p-p38MAPK단백표체균강저(P＜0.01);TNF-α단독간예조PEDF단백표체강저、p-p38MAPK단백표체증고(P＜0.01).결론 TA능구상조체외배양적인RPE세포PEDF단백표체,하조p-p38MAPK단백표체.
Objective To observe the influence of triamcinolone acetonide (TA) on the expression of pigment epithelium-derived factor (PEDF) of human retinal pigment epithelial (RPE) cells. Methods Cultured human RPE cells (4th - 6th generations) were treated with four different concentrations of TA (40, 400, 4× 103 and 4× 104 μg/L) for three different periods (12 or 24 or 48 hours), the levels of PEDF protein in the cell culture supernatant and cell lysates were determined by Western blot. After the initial experiment, RPE cells were treated with or without tumor necrosis factor-α (TNF-α, 20 ng/ml) for 24 hours, followed by TA (400 μg/L) treatment. The levels of PEDF and phospho-p38 mitogen activated protein kinase (p-p38MAPK) protein expression in cell culture supernatant and cell lysates were measured by Western blot. Results TA-treated RPE cells had higher PEDF expression, and 400 μg/L TA group had the highest effect (F= 16.98, P＜0. 05). 400μg/L TA treatment for one, six or 24 hours, with or without TNF-α pretreatment, could all promote the PEDF expression and inhibit the p-p38MAPK protein expression (F= 16.87, 10.28; P＜0. 01). TNF-α pretreatment alone could inhibit PEDF protein expression and promote p-p38MAPK protein expression (F= 16.87, 10. 28; P＜0. 01). Conclusions TA can up-regulate the expression of PEDF, and down-regulate the expression of p-p38MAPK in the cultured human RPE cells.
