眼科研究
眼科研究
안과연구
CHINESE OPHTHALMIC RESEARCH
2010年
1期
58-61
,共4页
晶状体上皮细胞%组织型转谷氨酰胺酶%细胞外基质%后囊膜混浊
晶狀體上皮細胞%組織型轉穀氨酰胺酶%細胞外基質%後囊膜混濁
정상체상피세포%조직형전곡안선알매%세포외기질%후낭막혼탁
lens epithelial cells%tissue transglutaminase%extracellular matrix,posterior capsule opacification
目的 观察组织型转谷氨酰胺酶(tTG)特异性抑制剂(MDC)对TGF-β_2诱导人晶状体上皮细胞(LECs)表达纤维连接蛋白(FN)、Ⅳ型胶原(Col-Ⅳ)的影响.方法 将含10%胎牛血清的DMEM培养基中培养的LECs株HLE-B3传代后分为5组:无血清培养基培养的HLE-B3为正常对照组;加入10μg/L TGF-β_2处理的HLE-B3为TGF-β_2组;10μg/L TGF-β_2与100、200、400μmol/L MDC共同处理的HLE-B3为不同浓度MDC组.用半定量RT-PCR检测tTG、FN、Col-Ⅳ在HLE-B3中的表达,计算A(tTG/β-actin)、A(FN/β-actin)、A(Col-Ⅳ/β-actin)值.结果 正常体外培养的HLE-B3中可表达一定量的tTG、FN及Col-Ⅳ.与正常对照组相比,10μg/L TGF-β_2组中tTG、FN、Col-Ⅳ的表达明显增加(t=33.95,P<0.01;t=38.24,P<0.01;t=13.48,P<0.01);与TGF-β_2组相比,100、200、400μmol/L MDC组中FN和Col-Ⅳ的表达明显减少(P<0.01).100、200、400μmol/L MDC组各组间FN和Col-Ⅳ的表达差异均有统计学意义(P<0.01).结论 MDC可剂量依赖性地抑制TGF-β_2诱导的LECs中FN、Col-Ⅳ表达的增加.tTG可促进人LECs表达FN、Col-Ⅳ,并参与后囊膜混浊(PCO)的形成.
目的 觀察組織型轉穀氨酰胺酶(tTG)特異性抑製劑(MDC)對TGF-β_2誘導人晶狀體上皮細胞(LECs)錶達纖維連接蛋白(FN)、Ⅳ型膠原(Col-Ⅳ)的影響.方法 將含10%胎牛血清的DMEM培養基中培養的LECs株HLE-B3傳代後分為5組:無血清培養基培養的HLE-B3為正常對照組;加入10μg/L TGF-β_2處理的HLE-B3為TGF-β_2組;10μg/L TGF-β_2與100、200、400μmol/L MDC共同處理的HLE-B3為不同濃度MDC組.用半定量RT-PCR檢測tTG、FN、Col-Ⅳ在HLE-B3中的錶達,計算A(tTG/β-actin)、A(FN/β-actin)、A(Col-Ⅳ/β-actin)值.結果 正常體外培養的HLE-B3中可錶達一定量的tTG、FN及Col-Ⅳ.與正常對照組相比,10μg/L TGF-β_2組中tTG、FN、Col-Ⅳ的錶達明顯增加(t=33.95,P<0.01;t=38.24,P<0.01;t=13.48,P<0.01);與TGF-β_2組相比,100、200、400μmol/L MDC組中FN和Col-Ⅳ的錶達明顯減少(P<0.01).100、200、400μmol/L MDC組各組間FN和Col-Ⅳ的錶達差異均有統計學意義(P<0.01).結論 MDC可劑量依賴性地抑製TGF-β_2誘導的LECs中FN、Col-Ⅳ錶達的增加.tTG可促進人LECs錶達FN、Col-Ⅳ,併參與後囊膜混濁(PCO)的形成.
목적 관찰조직형전곡안선알매(tTG)특이성억제제(MDC)대TGF-β_2유도인정상체상피세포(LECs)표체섬유련접단백(FN)、Ⅳ형효원(Col-Ⅳ)적영향.방법 장함10%태우혈청적DMEM배양기중배양적LECs주HLE-B3전대후분위5조:무혈청배양기배양적HLE-B3위정상대조조;가입10μg/L TGF-β_2처리적HLE-B3위TGF-β_2조;10μg/L TGF-β_2여100、200、400μmol/L MDC공동처리적HLE-B3위불동농도MDC조.용반정량RT-PCR검측tTG、FN、Col-Ⅳ재HLE-B3중적표체,계산A(tTG/β-actin)、A(FN/β-actin)、A(Col-Ⅳ/β-actin)치.결과 정상체외배양적HLE-B3중가표체일정량적tTG、FN급Col-Ⅳ.여정상대조조상비,10μg/L TGF-β_2조중tTG、FN、Col-Ⅳ적표체명현증가(t=33.95,P<0.01;t=38.24,P<0.01;t=13.48,P<0.01);여TGF-β_2조상비,100、200、400μmol/L MDC조중FN화Col-Ⅳ적표체명현감소(P<0.01).100、200、400μmol/L MDC조각조간FN화Col-Ⅳ적표체차이균유통계학의의(P<0.01).결론 MDC가제량의뢰성지억제TGF-β_2유도적LECs중FN、Col-Ⅳ표체적증가.tTG가촉진인LECs표체FN、Col-Ⅳ,병삼여후낭막혼탁(PCO)적형성.
BackgroundOur previous research and other reports disclosed that the expression of tissue transglutaminase(tTG)in lens epithelial cells(LECs) of patients with cataract is enhanced,indicating tTG is related to formation of posterior capsule opacification(PCO).ObjectivePresent study is to observe the effect of tTG specific inhibitor monodansyl-cadaverineon(MDC) on the expression of fibronectin(FN) and collagen Ⅳ(Col-Ⅳ) induced by TGF-β_2 in human LECs.MethodsHLE-B3 was cultured in vitro in DMEM containing 10% fetal bovine serum and then were divided into 5 groups.The free-serum culture was used as normal control group.Free-serum culture containing 10μg/L TGF-β_2 was utilized as treatment group.10μg/L TGF-β_2 plused 100μmol/L,200μmol/L and 400μmol/L MDC respectively in different concentrations as MDC-treatment group.Semiquantitative RT-PCR was used to assay the expression of tTG,FN and Col-Ⅳ in HLE-B3.A(tTG/β-actin),A(FN/β-actin) and A(Col-Ⅳ/β-actin) was calculated separately as the detecting indexes.ResultstTG,FN and Col-Ⅳ were positively expressed in cultured HLE-B3.The expression levels of tTG,FN and Col-Ⅳ in HLE-B3 were remarkably increased in the group with 10μg/L TGF-β_2 compared with normal control group(t=33.95,P<0.01;t=38.24,P<0.01;t=13.48,P<0.01).The expression levels of FN and Col-Ⅳ were gradually declined in 100,200 and 400μmol/L MDC groups in comparison with TGF-β_2 treatment(P<0.01).The significant differences were also found in the expressions of FN and Col-Ⅳ in HLE-B3 among 100,200 and 400μmol/L MDC groups(P<0.01).ConclusionMDC inhibits the expression of FN and Col-Ⅳ induced by TGF-β_2 in human LECs at a concentration-dependent manner.tTG may be involved in the formation of posterior capsule opacification through up-regulating the expressions of FN and Col-Ⅳ in human LECs.
