中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
Chinese Journal of Experimental Ophthalmology
2015年
11期
1004-1008
,共5页
李静%葛心%马建民%崔忆辛%李金茹%王霄娜
李靜%葛心%馬建民%崔憶辛%李金茹%王霄娜
리정%갈심%마건민%최억신%리금여%왕소나
眼眶炎性假瘤/药物治疗%成纤维细胞/药物作用%糖皮质激素/药学%地塞米松%细胞间黏附分子-1%下调%细胞培养
眼眶炎性假瘤/藥物治療%成纖維細胞/藥物作用%糖皮質激素/藥學%地塞米鬆%細胞間黏附分子-1%下調%細胞培養
안광염성가류/약물치료%성섬유세포/약물작용%당피질격소/약학%지새미송%세포간점부분자-1%하조%세포배양
Orbital pseudotumor/drug therapy%Fibroblasts/drug effects%Glucocorticoids/pharmacology%Dexamethasone%Intercellular adhesion molecule-1%Down-regulation%Cell culture
背景 特发性眼眶炎性假瘤(IOIP)是一种常见的眼眶疾病,病因和发病机制尚未完全阐明,以糖皮质激素为主的治疗效果欠佳. 目的 观察地塞米松对IOIP组织体外培养成纤维细胞的作用,探讨糖皮质激素在IOIP治疗中可能的作用机制.方法 收集2011年11月至2012年1月在北京同仁医院眼科中心切取的6例6眼IOIP组织(IOIP组)和3例因泪腺脱垂行泪腺复位术患者的眶部结缔组织(对照组),采用组织块培养法对2种组织进行体外培养以获取眼眶成纤维细胞,对培养的细胞进行组织形态学观察,采用免疫组织化学法检测培养细胞表面生物标志物的表达;采用WST-8法观察2个组培养的成纤维细胞的增生情况,并绘制生长曲线.将培养的细胞接种至96孔板,将不同浓度地塞米松(0、1×10-3、1×10-4、1×10-5和1×10-6mol/L)加入培养板分别作用24、48和72 h,采用WST-8法观察地塞米松对细胞增生的影响;采用ELISA法检测各组成纤维细胞中ICAM-1的相对表达量.结果 IOIP组织和正常眼眶体外培养的成纤维细胞均呈长梭形,两端有较长突起;免疫细胞化学染色可见2种组织中培养的细胞中vimentin呈阳性表达,而desmin、S-100、细胞角蛋白(CK)均呈阴性表达.与正常眼眶组织培养的成纤维细胞相比,IOIP组织培养的细胞生长速度较快.随着地塞米松浓度的增加,成纤维细胞的增生值(A值)逐渐下降,0、1 × 10-3、1 ×10-4、1×10-5和1×10-6mol/L地塞米松组成纤维细胞增生值的总体比较差异有统计学意义(F浓度=36.27,P=0.00),随着地塞米松作用时间的延长,成纤维细胞增生值逐渐下降,总体比较差异有统计学意义(F时间=3.69,P=0.00).无地塞米松培养组培养24、48和72 h,成纤维细胞中比较ICAM-1表达量分别为0.298±0.008、0.312±0.003和0.319±0.011,表现为逐渐升高的趋势,而不同浓度地塞米松作用后随着时间延长ICAM-1表达量均逐渐降低,总体比较差异有统计学意义(F时同=3.11,P=0.00),随着地塞米松浓度的增加,ICAM-1表达量均逐渐下降,总体比较差异有统计学意义(F浓度=75.17,P=0.00). 结论 IOIP的发生和发展可能与眼眶成纤维细胞中ICAM-1表达增强有关,地塞米松可能通过下调细胞中ICAM-1的表达而抑制眼眶成纤维细胞的增生,从而发挥抗炎和治疗作用.
揹景 特髮性眼眶炎性假瘤(IOIP)是一種常見的眼眶疾病,病因和髮病機製尚未完全闡明,以糖皮質激素為主的治療效果欠佳. 目的 觀察地塞米鬆對IOIP組織體外培養成纖維細胞的作用,探討糖皮質激素在IOIP治療中可能的作用機製.方法 收集2011年11月至2012年1月在北京同仁醫院眼科中心切取的6例6眼IOIP組織(IOIP組)和3例因淚腺脫垂行淚腺複位術患者的眶部結締組織(對照組),採用組織塊培養法對2種組織進行體外培養以穫取眼眶成纖維細胞,對培養的細胞進行組織形態學觀察,採用免疫組織化學法檢測培養細胞錶麵生物標誌物的錶達;採用WST-8法觀察2箇組培養的成纖維細胞的增生情況,併繪製生長麯線.將培養的細胞接種至96孔闆,將不同濃度地塞米鬆(0、1×10-3、1×10-4、1×10-5和1×10-6mol/L)加入培養闆分彆作用24、48和72 h,採用WST-8法觀察地塞米鬆對細胞增生的影響;採用ELISA法檢測各組成纖維細胞中ICAM-1的相對錶達量.結果 IOIP組織和正常眼眶體外培養的成纖維細胞均呈長梭形,兩耑有較長突起;免疫細胞化學染色可見2種組織中培養的細胞中vimentin呈暘性錶達,而desmin、S-100、細胞角蛋白(CK)均呈陰性錶達.與正常眼眶組織培養的成纖維細胞相比,IOIP組織培養的細胞生長速度較快.隨著地塞米鬆濃度的增加,成纖維細胞的增生值(A值)逐漸下降,0、1 × 10-3、1 ×10-4、1×10-5和1×10-6mol/L地塞米鬆組成纖維細胞增生值的總體比較差異有統計學意義(F濃度=36.27,P=0.00),隨著地塞米鬆作用時間的延長,成纖維細胞增生值逐漸下降,總體比較差異有統計學意義(F時間=3.69,P=0.00).無地塞米鬆培養組培養24、48和72 h,成纖維細胞中比較ICAM-1錶達量分彆為0.298±0.008、0.312±0.003和0.319±0.011,錶現為逐漸升高的趨勢,而不同濃度地塞米鬆作用後隨著時間延長ICAM-1錶達量均逐漸降低,總體比較差異有統計學意義(F時同=3.11,P=0.00),隨著地塞米鬆濃度的增加,ICAM-1錶達量均逐漸下降,總體比較差異有統計學意義(F濃度=75.17,P=0.00). 結論 IOIP的髮生和髮展可能與眼眶成纖維細胞中ICAM-1錶達增彊有關,地塞米鬆可能通過下調細胞中ICAM-1的錶達而抑製眼眶成纖維細胞的增生,從而髮揮抗炎和治療作用.
배경 특발성안광염성가류(IOIP)시일충상견적안광질병,병인화발병궤제상미완전천명,이당피질격소위주적치료효과흠가. 목적 관찰지새미송대IOIP조직체외배양성섬유세포적작용,탐토당피질격소재IOIP치료중가능적작용궤제.방법 수집2011년11월지2012년1월재북경동인의원안과중심절취적6례6안IOIP조직(IOIP조)화3례인루선탈수행루선복위술환자적광부결체조직(대조조),채용조직괴배양법대2충조직진행체외배양이획취안광성섬유세포,대배양적세포진행조직형태학관찰,채용면역조직화학법검측배양세포표면생물표지물적표체;채용WST-8법관찰2개조배양적성섬유세포적증생정황,병회제생장곡선.장배양적세포접충지96공판,장불동농도지새미송(0、1×10-3、1×10-4、1×10-5화1×10-6mol/L)가입배양판분별작용24、48화72 h,채용WST-8법관찰지새미송대세포증생적영향;채용ELISA법검측각조성섬유세포중ICAM-1적상대표체량.결과 IOIP조직화정상안광체외배양적성섬유세포균정장사형,량단유교장돌기;면역세포화학염색가견2충조직중배양적세포중vimentin정양성표체,이desmin、S-100、세포각단백(CK)균정음성표체.여정상안광조직배양적성섬유세포상비,IOIP조직배양적세포생장속도교쾌.수착지새미송농도적증가,성섬유세포적증생치(A치)축점하강,0、1 × 10-3、1 ×10-4、1×10-5화1×10-6mol/L지새미송조성섬유세포증생치적총체비교차이유통계학의의(F농도=36.27,P=0.00),수착지새미송작용시간적연장,성섬유세포증생치축점하강,총체비교차이유통계학의의(F시간=3.69,P=0.00).무지새미송배양조배양24、48화72 h,성섬유세포중비교ICAM-1표체량분별위0.298±0.008、0.312±0.003화0.319±0.011,표현위축점승고적추세,이불동농도지새미송작용후수착시간연장ICAM-1표체량균축점강저,총체비교차이유통계학의의(F시동=3.11,P=0.00),수착지새미송농도적증가,ICAM-1표체량균축점하강,총체비교차이유통계학의의(F농도=75.17,P=0.00). 결론 IOIP적발생화발전가능여안광성섬유세포중ICAM-1표체증강유관,지새미송가능통과하조세포중ICAM-1적표체이억제안광성섬유세포적증생,종이발휘항염화치료작용.
Background Idiopathic orbital inflammatory pseudotumor (IOIP) is a common orbital disease, but its etiology is still unclear,so the effect of glucocorticoid treatment is unsatisfied.Objective This study was to investigate the effects of dexamethasone on orbital fibroblasts from IOIP patients and explore the action machanism.Methods Six pieces of IOIP tissues from 6 IOIP patients and 3 pieces of normal orbital connective tissues from lacrimal gland prolapse patients were obtained during the surgery in Beijing Tongren Hospital from November 2011 to January 2012.The orbital fibroblasts were cultured using explant culture method.The morphology of the cells were observed under the optical microscope,and biomarks of the cells were detected by immunochemistry.The growth and proliferation of the cells were assayed using WST-8.The expression of ICAM-1 in the cells in both the control group and the IOIP group was detected by immunochemistry.The fibroblasts were incubated in 96-well plates, and different concentrations of dexamethasone (0,1 × 10-3 , 1 × 10-4 , 1 × 10-5 and 1 × 10-6 mol/L) were respectively added into the medium for 24,48 and 72 hours,and then the proliferation of the cells was detected by WST-8 assay.The contents of ICAM-1 in different concentrations of dexamethasone groups were assayed by ELISA.Results The characteristics of the cells were similar between the control group and the IOIP group with the spindle shape and long protructions.The cells showed the positive response for vimentin and absent response for desmin, S-100, cytokeratin (CK).Compared with the control group,the growth speed of fibroblasts was fast in the IOIP group.The proliferative values of the cells (absorbancy) were gradually reduced with the increase of dexamethasone concentrations (F ion =36.27,P=0.00) and the lapse of acting time (Ftime =3.69 ,P=0.00).In cultured cells without dexamethasone for 24,48 and 72 hours,the mean expression levels of ICAM-1 were 0.298±0.008,0.312±0.003 and 0.319±0.011, showing a gradually increasing trend.However,the expression of ICAM-1 was gradually reduced with the increases of concentrations and the lapse of acting time of dexamethasone (Fconcentration =75.17,P=0.00;Ftime =3.11,P=0.00).Conclusions Occurrence and development of IOIP is probably associated with the over-expression of ICAM-1 in orbital fibroblasts.Dexamethasone plays anti-inflammation and treating effects on IOIP by down-regulating the expression of ICAM-1 and inhibiting the proliferation of orbital fibroblasts.
