摘要
目的探讨炎症刺激对人脐带间充质干细胞生物学功能的影响及作用机制。方法无菌条件下取正常足月剖腹产脐带血40~60 ml,以Ficoll-Hypaque液梯度离心分离细胞,常规培养24 h后分为C组、LPSL组、LPSM组、LPSH组,C组加入完全培养液,LPSL组、LPSM组、LPSH组分别加入含有12.5、25、50μg/ml脂多糖(LPS)的等量培养液,观察不同浓度LPS孵育对脐带间充质干细胞Toll样受体4(TLR4)、核因子-κB(NF-κB)蛋白表达间充质干细胞增殖和成骨分化的影响。结果孵育24 h和48 h,LPSL、LPSM、LPSH组间充质干细胞增殖活性均高于C组(P<0.05),且低剂量LPS诱导间充质干细胞的增殖能力最强(P<0.05),48 h增殖活性高于24 h(P<0.05)。随着培养时间增加,四组NF-κB、TLR4蛋白表达逐渐增加,LPSL、LPSM、LPSH组均高于C组(P<0.05),且随着LPS浓度升高,NF-κB、TLR4蛋白表达逐渐升高,差异均有统计学意义(P<0.05)。LPSL、LPSM、LPSH组碱性磷酸酶(ALP)活力和钙结节数量均高于C组,LPSM组ALP活力和钙结节数量高于LPSH组,LPSL组ALP活力和钙结节数量高于LPSM组和LPSH组,差异均有统计学意义(P<0.05)。结论炎性反应可刺激人脐带间质干细胞增殖和成骨分化,低浓度的LPS可能是脐带间充质干细胞成骨分化的理想因子,在实际的临床疾病治疗中具有一定的意义和应用前景。
Objective To investigate the effect of inflammatory stimulation on the biological function of human umbilical cord mesenchymal stem cells(MSCs)and its mechanism.Methods Under sterile condition,40-60 ml cord blood of normal full-term caesarean section was taken,and the cells were separated by Ficoll Hypaque gradient centrifugation.After 24 hours of conventional culture,the cells were divided into 4 groups:C group(control group),LPSL group,LPSM group and LPSH group.Group C was added with complete culture medium.Group LPSL,group LPSM and group LPSH were added with equal amount of culture medium containing 12.5,25,50μg/ml lipopolysaccharides(LPS)respectively.The effects of different concentrations of LPS on the proliferation and osteogenic differentiation of mesenchymal stem cells were observed.Results After incubation for 24 and 48 hours,the proliferation activity of MSCs in LPSL,LPSM and LPSH groups was higher than that in group C(P<0.05),and the proliferation ability of MSCs induced by low dose LPS for 48 h was the strongest(P<0.05).With the increase of culture time,the expression of nuclear factor kappa B(NF-κB)and Toll-like receptor 4(TLR4)protein in the four groups increased gradually.The expression of NF-κB and TLR4 protein in the LPSL,LPSM and LPSH groups were higher than that in the control group(P<0.05).With the increase of LPS concentration,the expression of NF-κB and TLR4 protein increased gradually,with statistically significant difference(P<0.05).Alkaline phosphatase(ALP)activity and calcium nodule number in LPSL,LPSM and LPSH group were higher than those in group C;ALP activity and calcium nodule number in LPSM group were higher than those in LPSH group;ALP activity and calcium nodule number in LPSL group were higher than those in LPSM and LPSH,group with statistically significant difference(P<0.05).Conclusions Inflammatory response can stimulate the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells.Low concentration of LPS may be an ideal factor for the osteogenic differentiation of umbilical cord mesenchymal stem cells.It has certain significance and application prospect in the actual clinical disease treatment,but the detailed mechanism of action and whether the effect of lower dose is better still to be further studied.
作者
王晔
李俊玲
季珊
王伟
李艳
Wang Ye;Li Junling;Ji Shan;Wang Wei;Li Yan(No.2 Newborn Ward,Baoding Maternal and Child Health Care Hospital,Baoding 071000,China;Department of Medical Record,Baoding Maternal and Child Health Hospital,Baoding 071000,China)
出处
《中国医师杂志》
CAS
2019年第12期1825-1828,共4页
Journal of Chinese Physician
关键词
脂多糖类
脐带
间充质基质细胞
细胞增殖
体外研究
Lipopolysaccharides
Umbilical cord
Mesenchymal stromal cells
Cell proliferation
In vitro
