Chen H-T, Huang A-B, He Y-L, Bian J, Li H-J
Department of Orthopedics, Taizhou People's Hospital Affiliated to Nantong University, Taizhou, Jiangsu, China.
Eur Rev Med Pharmacol Sci. 2017 Apr;21(7):1462-1470.
Our present study aimed to evaluate the effects of Wnt11 overexpression on the adipose-derived stem (ADSCs) cells differentiation to the nucleus pulposus (NP) cells and its function in the ADSCs cells growth, proliferation and induction of the NP cells markers.
The cell growth was detected using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (MTT) assay and the cell cycle was assessed by the flow cytometry. The cells morphology was evaluated using the transmission electron microscopy. The transfection efficiencies of Wnt11 lentivirus were observed under fluorescence microscope. Besides, Quantitative Real-time PCR and Western blot analysis were applied to detect the relative mRNA and protein levels.
Wnt11 lentivirus treatment could inhibit the ADSCs cells growth and arrest the cell cycle progression at the G0/G1 phase. Besides, the overexpression of Wnt11in ADSCs cells could induce the expression of the NP cells markers. Levels of SOX-9, aggrecan, and collagen type II were significantly increased in the ADSCs cells transfected with the Wnt11 lentivirus, in comparison with the untreated cells or the vector controls.
The Wnt11 overexpression may provide some experimental evidence for the possible opportunity of the Wnt11 to promote the ADSCs cells differentiating to the NP cells. Therefore, the Wnt11 overexpression may have a potential utility for the treatment of the intervertebral disc degeneration.
本研究旨在评估Wnt11过表达对脂肪来源干细胞(ADSCs)向髓核(NP)细胞分化的影响及其在ADSCs细胞生长、增殖和诱导NP细胞标志物方面的作用。
采用(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐)(MTT)法检测细胞生长情况,通过流式细胞术评估细胞周期。使用透射电子显微镜评估细胞形态。在荧光显微镜下观察Wnt11慢病毒的转染效率。此外,应用定量实时PCR和蛋白质印迹分析检测相对mRNA和蛋白质水平。
Wnt11慢病毒处理可抑制ADSCs细胞生长并使细胞周期停滞在G0/G1期。此外,ADSCs细胞中Wnt11的过表达可诱导NP细胞标志物的表达。与未处理细胞或载体对照相比,用Wnt11慢病毒转染的ADSCs细胞中SOX-9、聚集蛋白聚糖和II型胶原的水平显著升高。
Wnt11过表达可能为Wnt11促进ADSCs细胞向NP细胞分化的潜在机会提供一些实验证据。因此,Wnt11过表达可能对椎间盘退变的治疗具有潜在应用价值。