摘要
目的探究原花青素(PACs)对牙周膜干细胞(PDLSCs)增殖、凋亡与成骨分化诱导的影响及相关机制。方法将体外培养PDLSCs细胞分为Ctl组、2.5、5、10、20、40 mg·L^(-1) PACs组、SP600125组、10 mg·L^(-1) PACs+SP600125组,采用CCK-8、5-乙基-2’-脱氧尿苷法、Hoechst 33258染色法、碱性磷酸酶(ALP)活性测定法、茜素红S染色(ARS)法、蛋白免疫印迹法分析细胞活力、增殖、凋亡、ALP活性、细胞矿化结节及蛋白表达。结果24 h后,与Ctl组比较,10 mg·L^(-1) PACs组细胞增殖率升高(P<0.05);诱导7 d后,与Ctl组比较,2.5、5、10 mg·L^(-1) PACs组成骨标志物骨桥蛋白(OPN)、骨唾液蛋白II(BSP2)、Osterix(OSX)、ALP及p-JNK表达水平上调(P<0.05),5、10 mg·L^(-1) PACs组ALP水平升高(P<0.05);骨诱导14 d后,各组均形成橙色钙,5、10 mg·L^(-1) PACs组ARS水平高于Ctl组(P<0.05)。SP600125组细胞增殖率、ALP活性、ARS及BSP2、OSX、p-JNK表达低于Ctl组(P<0.05);10 mg·L^(-1) PACs+SP600125组细胞增殖率、ALP活性、ARS、OPN、BSP2、OSX、ALP及p-JNK表达水平低于10 mg·L^(-1) PACs组,高于SP600125组(P<0.05)。结论PACs可通过上调JNK信号通路,促进PDLSCs细胞活力、增殖及诱导成骨分化。
Objective To investigate the effects of proanthocyanidins(PACs)on the proliferation,apoptosis and osteogenic differentiation induction of periodontal ligament stem cells(PDLSCs)and the related mechanisms.Methods PDLSCs cultured in vitro were divided into the control group,2.5,5,10,20,40 mg·L^(-1) PACs group,SP600125 group,and 10 mg·L^(-1) PACs+SP600125 group.CCK-8,5-ethynyl-2’-deoxyuridine assay,Hoechst 33258 staining,alkaline phosphatase(ALP)activity assay,alizarin red S staining(ARS)and Western blot were used to analyze the cell viability,proliferation,apoptosis,ALP activity,cell mineralized nodules and protein expression.Results After 24 hours,the cell proliferation rate of the 10 mg·L^(-1) PACs group was increased compared with the control group(P<0.05).After 7 days of induction,compared with control group,the expression levels of osteogenic markers osteopontin(OPN),bone sialprotein II(BSP2),Osterix(OSX),ALP and p-JNK in 2.5,5,10 mg·L^(-1) PACs groups were up-regulated(P<0.05),while the levels of ALP in 5 and 10 mg·L^(-1) PACs groups were increased(P<0.05).After 14 days of osteoinduction,orange calcium was formed in all groups,and the ARS levels in 5 and 10 mg·L^(-1) PACs groups were higher than those in control group(P<0.05).The cell proliferation rate,ALP activity,ARS,BSP2,OSX,p-JNK expression in SP600125 group were lower than those in the control group(P<0.05).The cell proliferation rate,ALP activity,ARS,OPN,BSP2,OSX,ALP and p-JNK expression levels in 10 mg·L^(-1) PACs+SP600125 group were lower than those in 10 mg·L^(-1) PACs group and higher than those in SP600125 group(P<0.05).Conclusion PACs can promote the cell viability,proliferation and the inducible osteogenic differentiation of PDLSCs through up-regulating JNK signaling pathway.
作者
曾华
曹素敏
李召宝
ZENG Hua;CAO Sumin;LI Zhaobao(Department of Stomatology,Xiamen Xianyue Hospital/Xianyue Hospital Affiliated to Xiamen Medical College/Fujian Psychiatric Center/Fujian Clinical Research Center for Mental Disorders,Xiamen 361000,China;Department of Stomatology,Cangzhou Central Hospital,Cangzhou 061000,China)
出处
《长春中医药大学学报》
2025年第2期160-167,共8页
Journal of Changchun University of Chinese Medicine
基金
2021年度河北省医学科学研究课题计划项目(20210670)。
关键词
原花青素
牙周膜干细胞
成骨分化
C-JUN氨基端激酶
增殖
proanthocyanidins
periodontal ligament stem cells
osteogenic differentiation
c-Jun N-terminal kinase
proliferation
