ABSTRACT
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.
ABSTRACT
The effects of lutein on the growth and migration of bovine lens epithelial cells (BLECs) in vitro were observed in an attempt to find a drug that can prevent after-cataract. BLECs were cultured in vitro and different concentrations of lutein were added to the BLECs cultures of the second and third generations. The effects of lutein on the proliferation of BLECs in vitro were examined by the MTT method, and the migration of BLECs was evaluated by a scratch wound assay. The results showed that: (1) Lutein at concentrations of 1 to 16μmol/L could inhibit the proliferation of BLECs in a dose-and time-dependent manner (P<0.01); (2) The migration of BLECs was evaluated by wound healing rate. As compared with the control group, the wound healing rate in the experimental groups was decreased from 0.672±0.164 to -0.234±0.144 and -0.597±0.063 (P<0.01) at 1 and 2μmol/L lutein, respectively. It was concluded that lutein at concentration of ≥1μmol/L inhibited the proliferation and migration of BLECs in vitro. Lutein may become an effective drug to prevent after-cataract.
ABSTRACT
To investigate the effect of proteasome inhibitor MG132 on the apoptosis of bovine lens epithelial cells (BLECs), the cells were treated with MG132 at different concentrations for12, 24 and 36 h. The cell viability was analyzed by MTT assay and the effect of MG132 on the apoptosis of BLECs was assessed by flow cytometry (FCM). The results showed that after treatment for the same period, the inhibitory effect of MG132 on BLECs proliferation was enhanced with the increment of the concentration of MG132 (0, 2, 5, 10, μmol/L) (P＜0.05). The 50% inhibiting concentration (IC50)was 2.03 μmol/L when the BLECs were treated with MG132 for 36 h. MG132 also induced the apoptosis of BLECs obviously. FCM showed that the apoptosis index of the cells treated by MG132 at 2μmol/L for 12 h was (20.24±1.51)%, and that of the control was (0.98±0.20)% respectively(P＜0.01, n=3). It was concluded that MG132 could lead to apoptosis of BLECs. The decrease of proteasome activity may play an important role in the formation and development of cataract.