AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfect...AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfected with pEGFP-Msx2 and control. Msx2-deficient mice (Msx2(-/-)) lens tissue were isolated. Lens tissue and transfected cells were prepared for mRNA extraction using Trizol reagent. CyclinD1 and Prox1 expression were evaluated by real-time RT-PCR. BrdU incorporation and apoptosis rate were investigated by immunofluorescence and flow cytometry analysis. RESULTS: After transfected with pEGFP-Msx2, lens epithelium cells failed to incorporate BrdU and anti - phospho-histone-3 immunofluorescence failed to detect cell nuclei which GFP were positive. Msx2 over expression resulted in increasing apoptosis rate in lens epithelium cells. CyclinD1 and Prox1 expression increased significantly in Msx2 knockout mice by real time RT-PCR quantization and CyclinD1 expression decreased significantly in Msx2overexpressed cell. CONCLUSION: Msx2 has the effect of inhibiting proliferation and differentiation, triggering apoptosis on mice lens epithelium cells.展开更多
AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition (EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery (FLACS). METHODS: Sixty cataract ...AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition (EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery (FLACS). METHODS: Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly: FLACSl group (cataract surgery by FLACS with LenSx), FLACS2 group (cataract surgery by FLACS with LensAR) and manual group (cataract surgery by phacoemulsification). Patients in two FLACS groups performed anterior capsulotomy by LenSx or LensAR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis (CCC) manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery. RESULTS: The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC (P〈0.05). Lens epithelium cells apoptosis were correlated with femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT. CONCLUSION: Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.展开更多
Purpose:To study the ability of Homoharringtonine(Hh),5-Fluorouracil(5-Fu).and Adriamycin(ADM)on inhibiting the proliferation of rabbit lens epitthelium,Methods.Whole rabbit lenses were removed from freshly enucleated...Purpose:To study the ability of Homoharringtonine(Hh),5-Fluorouracil(5-Fu).and Adriamycin(ADM)on inhibiting the proliferation of rabbit lens epitthelium,Methods.Whole rabbit lenses were removed from freshly enucleated eyes under sterile condition.The rabbit lens eptithlia(RLE)were isolated and culatured:(1)The passage RLE were placed in 24-well tissue culture plates and incubated for 48hours.then exposed to different concentrations of Hh,5-Fu,and ADMfor 24and 72hours;(2)The passage RLEandHh(0.084μg/ml).5-Fu(0.058μg/ml),ADM(0.45ng/ml)were placed and cultured for 24hours;(3)The morphological changes of RLE exposed to different concentrations of Hh,5-Fu and ADM were studied under light microscope.Results:The ID50 of Hh,5-Fu and ADMexposed to RLEfor 24hours were 0.84μg/ml, 0.58μg/ml and4.50ng/ml,respectively,and those for 72hous were0.49μg/ml,0.33μg/ml and3.85ng/ml.The attachment rate of RLE after being cu-latured for 24hours with Hh,5-Fu and ADM were respectinely83.6%,89.1%and 87.3%,The morphological changes of RLE demonstrated that obvious changes in the cell membran e and cytoplasm were found even in lower concentra-tion ,but changes in the nuclei could only be found in higher concentation of these drugs.Conclusion:Hh can not only inhibit the proliferation of RLE but also reduce the number of attached cells.It is suggested that Hh may be more useful for the pre-vention of after cataract than 5-Fu and ADM.展开更多
基金Science Foundation of Liaoning Province,China (No. 2011225014)
文摘AIM: To investigate the effects of Msx2 on lens epithelium cell cycle, and evaluate the changes of the proliferation, apoptosis of lens epithelium cells. METHODS: Mice lens epithelium cells were cultured and transfected with pEGFP-Msx2 and control. Msx2-deficient mice (Msx2(-/-)) lens tissue were isolated. Lens tissue and transfected cells were prepared for mRNA extraction using Trizol reagent. CyclinD1 and Prox1 expression were evaluated by real-time RT-PCR. BrdU incorporation and apoptosis rate were investigated by immunofluorescence and flow cytometry analysis. RESULTS: After transfected with pEGFP-Msx2, lens epithelium cells failed to incorporate BrdU and anti - phospho-histone-3 immunofluorescence failed to detect cell nuclei which GFP were positive. Msx2 over expression resulted in increasing apoptosis rate in lens epithelium cells. CyclinD1 and Prox1 expression increased significantly in Msx2 knockout mice by real time RT-PCR quantization and CyclinD1 expression decreased significantly in Msx2overexpressed cell. CONCLUSION: Msx2 has the effect of inhibiting proliferation and differentiation, triggering apoptosis on mice lens epithelium cells.
文摘AIM: To evaluate human lens epithelium cell apoptosis and epithelial to mesenchymal transition (EMT) induced by femtosecond laser in femtosecond laser assisted cataract surgery (FLACS). METHODS: Sixty cataract patients with N2 to N3 stage according to the LOCS III were enrolled in this study and divided into three groups randomly: FLACSl group (cataract surgery by FLACS with LenSx), FLACS2 group (cataract surgery by FLACS with LensAR) and manual group (cataract surgery by phacoemulsification). Patients in two FLACS groups performed anterior capsulotomy by LenSx or LensAR laser system. Patients in the manual group were performed continuous curvilinear capsulorrhexis (CCC) manually. The anterior capsules were fixed right after moved out of eye. Hematoxylin-eosine staining, immunofluorescence staining and real-time PCR were performed in order to observe human lens epithelium cells changes after cataract surgery. RESULTS: The capsule cutting edge was shown irregularity and roughness in two FLACS groups and smooth edge in manual capsulotomy by pathologic staining. Irregularities of the cell configuration with partly swollen and destroyed nuclei were observed in two FLACS groups. Femtosecond laser could induce a significantly higher cell apoptosis in human lens epithelium cell than manually performed CCC (P〈0.05). Lens epithelium cells apoptosis were correlated with femtosecond laser duration according to Pearson correlation analysis. Decreased N-cadherin expression, alpha-SMA and FSP-1 level in two FLACS groups showed the inhibition of cell EMT. CONCLUSION: Femtosecond laser may affect the apoptosis and EMT of lens epithelium cells which are under the peeled central lens capsule.
文摘Purpose:To study the ability of Homoharringtonine(Hh),5-Fluorouracil(5-Fu).and Adriamycin(ADM)on inhibiting the proliferation of rabbit lens epitthelium,Methods.Whole rabbit lenses were removed from freshly enucleated eyes under sterile condition.The rabbit lens eptithlia(RLE)were isolated and culatured:(1)The passage RLE were placed in 24-well tissue culture plates and incubated for 48hours.then exposed to different concentrations of Hh,5-Fu,and ADMfor 24and 72hours;(2)The passage RLEandHh(0.084μg/ml).5-Fu(0.058μg/ml),ADM(0.45ng/ml)were placed and cultured for 24hours;(3)The morphological changes of RLE exposed to different concentrations of Hh,5-Fu and ADM were studied under light microscope.Results:The ID50 of Hh,5-Fu and ADMexposed to RLEfor 24hours were 0.84μg/ml, 0.58μg/ml and4.50ng/ml,respectively,and those for 72hous were0.49μg/ml,0.33μg/ml and3.85ng/ml.The attachment rate of RLE after being cu-latured for 24hours with Hh,5-Fu and ADM were respectinely83.6%,89.1%and 87.3%,The morphological changes of RLE demonstrated that obvious changes in the cell membran e and cytoplasm were found even in lower concentra-tion ,but changes in the nuclei could only be found in higher concentation of these drugs.Conclusion:Hh can not only inhibit the proliferation of RLE but also reduce the number of attached cells.It is suggested that Hh may be more useful for the pre-vention of after cataract than 5-Fu and ADM.