The Effect Of TSP1 In The Formation Of Anterior Subcapsular Cataract

Objective: To explore the expression and significance of thrombospondin 1(TSP1) in human lens epithelial cells (HLECs) and cultured monkey lense invitro.Furthermore,we investigate whether TSP1 induces anterior subcapsular cataract.Methods: TSP1 was expressed using pGEX4T1 vector in E.coli prokaryoticexpression system, purified via high affinity chromatography and subjected toWestern Blot analysis. Purified TSP1 was added to media containing cultured monkeylenses, and the morphological changes of the lenses were studied. In addition, humanlens epithelial cells (HLECs) were infected with adenoviral vectors containing TSP1,and the morphological changes of HLECs were also studied. Last, TSP1 proteinpurchased was added to media containing cultured monkey lenses, and themorphological changes of the lenses were evaluated.Results: Western blot analysis did not reveal a 154kD band as expected,suggesting TSP1 is not suitable for prokaryotic expression. Both empty adenoviralvector and TSP1packed adenoviral vector induced elongation of HLECs andincreased intercellular space, but the latter showed more dramatic HLEC elongationeffect. Lenses cultured with TSP1 developed distinct anterior opacities just beneaththe lens capsule.Conclusion: TSP1 may induce cellular elongation in HLEC and possibly induceanterior subcapsular cataract.