ABSTRACT
Objective:To investigate the effects of Dickkopf-1(DKK1) on the proliferation of human lens epithelial cells (LECs) and its possible mechanism in order to search a new target for the treatment of posterior capsular opacification (PCO).Methods:Human LECs line (SRA 01/04 cells )were divided into Wnt3a overexpression group, DKK1 group and control group.Wnt3a gene expression vector was transfected into SRA 01/04 cells by liposome mediated transfection to establish a PCO model in the Wnt3a overexpression group, DKK1 of 100 μg/ml was added into the medium 48 hours after transfection of Wnt3a gene vector in the DKK1 group, and only pcDNA3-HA vector was transfected in the control group.The survival rate of SRA 01/04 cells was detected with a cell counting kit-8 (CCK-8) assay.The expression rate of proliferating cell nuclear antigen (PCNA) in the cells was detecteds by immunocytochemistry.The expression of β-catenin in the cells was detected and located by immunofluorescence.The expression of Wnt3a CyclinD1 and C-Myc were detected by Western blot assay.Results:The relative expression of Wnt3a protein in the control group was 0.49±0.07, which was significantly lower than that in the Wnt3a overexpression group (0.84±0.06) ( t=3.704, P=0.02). The survival rate in the Wnt3a overexpression group, DKK1 group and control group showed significant difference over time ( Fgroup=10.910, P<0.05; Ftime=6.041, P<0.05). The survival rate in the Wnt3a overexpression group was significantly increased in comparison with the control group and that in the DKK1 group was significantly reduced in comparison with the Wnt3a overexpression group(all at P<0.05). β-Catenin was expressed mainly in cytoplasm and cell nucleus in the Wnt3a overexpression group and only in cytoplasm in the DKK1 group. in the control group, β-catenin showed a weaked expression in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The expression rates of PCNA protein were (9.4±1.4)%, (43.4±5.4)%, and (14.2±2.3)% in the control group, Wnt3a overexpression group and DKK1 group, respectively, with a significant difference among the groups ( F=28.250, P<0.05), and the expression rates of PCNA protein were significantly reduced in the control group and DKK1 group compared with the Wnt3a overexpression group (both at P<0.05). β-Catenin protein were expressed mainly in the cytoplasm and nucleus in the Wnt3a overexpression group and only in the cytoplasm in the control group.In the DKK1 group, the expression of β-catenin protein was weakened in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The relative expressions of CyclinD1 were 0.64±0.07、0.84±0.03 and 0.55±0.10, C-Myc were 0.59±0.05、0.93±0.02 and 0.47±0.08 in the control group, Wnt3a overexpression group and the DKK1 group, respectively with significant differences among the groups ( F=20.580, 5.040, both at P<0.05). The relative expressions of CyclinD1 and C-Myc in the Wnt3a overexpression group were significantly higher than those in the DKK1 group and the control group (all at P<0.05). Conclusions:DKK1 inhibits activation of Wnt/β-catenin signaling pathway induced by Wnt3a overexpression in SRA01/04 cells, and down-regulation of downstream target proteins cyclin D1 and C-Mgc may be the biological mechanism of Dkk1 inhibiting human LECs proliferation.
ABSTRACT
Objective To discuss the perioperative nursing measures for patients with osteoporotic vertebral compression fractures who are receiving percutaneous vertebroplasty (PVP) treatment by using high viscosity bone cement.Methods A total of 30 patients with osteoporotic vertebral compression fractures were included in this study.All patients were treated with PVP by using high viscosity bone cement.Preoperative routine nursing,psychological intervention,dietary intervention,postoperative guidance of rehabilitation exercise of limbs,close observation of bone cement leakage were strictly implemented,and the corresponding nursing measures were promptly taken when needed.Results Through strict implement of the nursing intervention all 30 patients could actively cooperate with PVP treatment,and after PVP the pain was significantly relieved in all patients.Conclusion Adequate preoperative preparation,proper postoperative guidance,careful observation and effective nursing can help the patients resume their daily life activities as soon as possible,relieve the pain,and improve the quality of life as well.(J Intervent Radiol,2017,26:274-276)
ABSTRACT
Background Excessive production,deposition and contraction of extracellular matrix (ECM) of human lens endothelial cells (LECs) is one of main causes to posterior capsular opacification (PCO).Researches indicated that Wnt3a protein was involved in production of ECM and fibrosis in epithelial cells,but its effect on LECs is still unclear.Objective This study aimed to elucidate the roles of Wnt3a in production of ECM and gel contraction of LECs.Methods Lipofectamine-mediated transient transfection technique was used to introduce cDNA of Wnt3a gene and pcDNA3-HA vector into the LEC cell line SRA01/04 to act as Wnt3a transfection group and control group respectively.After 48 h of transfection,Western blot was used to detect the expression of Wnt3a,main components of ECM Col-Ⅰ,Col-Ⅳ and integrin β1.Immunofluorescence was used to detect the expression and distrubition of α-SMA and F-actin;collagen contraction was observed by mingling SRA01/04 cells with Col-Ⅰ.Results After 48 hours of transfection using lipofectamine 2000,the expressions of wnt3a protein in SRA01/04 cells was 0.703 ±0.105 in the wnt3a transfected group,and that in the control group was 0.290 ± 0.066,showing a significant difference between the two groups (t =5.782,P＜0.01).Western blot assay showed that the expression levels of Col-Ⅰ and Integrin β1 was 0.697±0.021 and 0.875±0.055 in the Wnt3a transfected group,and which was significantly higher than 0.370±0.020 and 0.580±0.030 in the control group (t =19.600,8.156,both P＜0.01).The expression level of Col Ⅳ in the Wnt3a transfected group was higher than that of the control group (0.430±0.020 vs 0.383 ±0.031),but the difference was not significant (t =2.514,P＞0.05).Immunofluorescence assay revealed that F-actin and α-SMA were weakly expressed in the cell membrane primarily in the control group,while they were strongly expressed in the cell membrane,cytoplasma in the Wnt3a transfected group.Tewnty-four hours after addtion of Col-Ⅰ,the gel contraction area ratio appeared to be more obvious in comparison with the 8 hours (64.1％ ±2.3％ vs 98.9％ ± 1.0％),and gel contraction area ratio was lower in the Wnt3a transfected group than that in the control group (64.1％ ± 2.3％ vs 93.9％ ± 3.1％).Conclusions The overexpression of Wnt3a activates the production of ECM,and the remodeling of celluar skeleton and cellular contraction.