牙龈卟啉单胞菌感染通过Wnt通路调节牙周膜干细胞的成骨分化  

The regulatory effect of Porphyromonas gingivalis infection on osteogenic differentiation of periodontal ligament stem cells through Wnt pathway

作　　者：张小松 陈红莉[2] 王志刚[2] ZHANG Xiao-song;CHEN Hong-li;WANG Zhi-gang(Department of Stomatology,the Third People's Hospital Directly Subordinate to Henan Province,Zhengzhou,Henan 450000,China;不详)

机构地区：[1]河南省直第三人民医院口腔科,河南郑州450000 [2]河南省人民医院口腔医学中心,河南郑州450000

出　　处：《中国微生态学杂志》2021年第1期47-50,共4页Chinese Journal of Microecology

基　　金：郑州市医学科技攻关计划项目(201404032)。

摘　　要：目的观察牙龈卟啉单胞菌(P.gingivalis)感染通过Wnt通路调节牙周膜干细胞(PDLSCs)成骨分化的作用。方法培养原代PDLSCs,分为常规处理的对照组、P.gingivalis感染的P.gingivalis组和P.gingivalis感染并用Wnt3a处理的P.gingivalis+Wnt3a组,成骨诱导后茜素红染色并检测A405值,Western blot检测Wnt通路分子的蛋白表达量,碱性磷酸酶(ALP)试剂盒检测ALP活力,PCR检测成骨标志基因Runt相关转录因子2(Runx2)、骨钙素(OCN)的mRNA表达量。结果与对照组比较,P.gingivalis组Wnt3a、β-catenin、p-GSK-3β的蛋白表达水平(0.33±0.07)、(0.27±0.08)、(0.44±0.09)以及成骨诱导后A405值(0.55±0.08)、ALP活力(20.14±6.54)U/mL和Runx2、OCN的mRNA表达量(0.45±0.09)、(0.51±0.07)均明显减少;与P.gingivalis组比较,P.gingivalis+Wnt3a组成骨诱导后A405值(0.89±0.15)、ALP活力(29.44±5.26)U/mL及Runx2、OCN的mRNA表达量(0.89±0.17)、(0.81±0.18)均明显增加。结论P.gingivalis感染能够抑制PDLSCs的成骨分化,抑制Wnt通路是可能的分子机制。Objective To observe the effect of P.gingivalis infection on osteogenic differentiation of periodontal ligament stem cells(PDLSCs)through Wnt pathway.Methods Primary PDLSCs were cultured and divided into control group,P.gingivalis group,or P.gingivalis+Wnt3 a group.After osteogenesis induction,alizarin red staining was performed and A405 were detected.The protein expression of Wnt pathway molecule was detected using Western blot,ALP activity was detected with ALP kit,and the mRNA expression of Runt-related transcription factor 2(RUNX2)and osteocalcin(OCN)were detected by using PCR.Results Compared with the control group,the protein expression levels of Wnt3 a,β-Catenin and p-GSK-3βwere(0.33±0.07),(0.27±0.08),(0.44±0.09),the A405 level(0.55±0.08),ALP activity(20.14±6.54)U/mL,and mRNA expression of Runx2 and OCN(0.45±0.09),(0.51±0.07)in P.gingivalis group significantly reduced;Compared with P.gingivalis group,the A405 level(0.89±0.15),ALP activity(29.44±5.26)U/mL,and mRNA expression of Runx2 and OCN(0.89±0.17),(0.81±0.18)in P.gingivalis+Wnt3 a group significantly increased.Conclusion P.gingivalis infection can inhibit the osteogenic differentiation of PDLSCs,and the inhibition of Wnt pathway is possibly the molecular mechanism.

关 键 词：牙龈卟啉单胞菌 牙周膜干细胞 成骨分化 WNT通路 

分 类 号：R781.4[医药卫生—口腔医学]