Leptin转染人胎盘间充质干细胞对辐照后口腔黏膜修复细胞的作用及机制分析  

Effect and mechanism of leptin transfected human placental mesenchymal stem cells on oral mucosa repair cells after irradiation

作　　者：杨雨 刘定坤 李凌峰 隋欣 王碧舟 刘志辉[1] YANG Yu;LIU Ding-kun;LI Ling-feng(Stomatological Hospital of Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学口腔医院,吉林长春130021

出　　处：《中国实验诊断学》2021年第12期1825-1830,共6页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省重点科技研发项目(20180201056YY);吉林省教育厅“十三五”科学技术项目(JJKH20180236KJ);长春市地院合作专项(17DY024)。

摘　　要：目的探讨瘦素(leptin)转染的人胎盘间充质干细胞(HPMSCs)对被X射线辐照后的口腔黏膜修复细胞的作用影响,为进一步的临床治疗提供新思路。方法实验分为转染leptin的HPMSCs组(HPMSCs/Leptin)和转染空载体的HPMSCs组(HPMSCs/NC),将人牙龈成纤维细胞(HGnF)、人口腔角质细胞(HOK)和人脐静脉内皮细胞(HUVEC)进行单剂量20Gy的X射线照射,Transwell检测3种被X射线辐照后细胞的迁移能力。构建HPMSCs/Leptin或HPMSCs/NC与辐照后HGnF、HOK和HUVEC的共培养体系,通过CCK-8检测被X射线辐照后3种细胞在24h、48h、72h、96h的增殖活性,ELISA检测被X射线辐照后3种细胞在24h、48h、72h表达炎症因子(人IL-1α、IL-1β、IL-6、IL-8、GM-CSF和TNF-α)的水平。结果 HPMSCs/Leptin可促进HUVEC的迁移,抑制HGnF和HOK的迁移。HPMSCs/Leptin能增强辐照损伤后的HOK和HUVEC的增殖能力,但对HGnF的增殖影响不明显。HPMSCs/Leptin能促进HGnF、HOK和HUVEC分泌炎症因子人IL-1α、IL-6、GM-CSF和TNF-α,但对人IL-1β和IL-8的分泌无明显影响。结论瘦素转染的人胎盘源间充质干细胞HPMSCs/Leptin可通过补充口腔黏膜修复细胞数量、调节炎症反应、促进血管新生等途径,对促进放创复合伤伤口愈合过程发挥一定的积极作用。Objective To investigate the effect of leptin-transfected human placental mesenchymal stem cells(HPMSCs)on oral mucosa repair cells irradiated by X-ray,which provides new ideas for clinical treatment.Methods The experiment was divided into HPMSCs group(HPMSCs/Leptin)transfected with leptin and HPMSCs group(HPMSCs/NC)transfected with empty vector.Human gingival fibroblasts(HGnF),human oral keratinocytes(HOK)and Human umbilical vein endothelial cells(HUVEC)were irradiated with a single dose of 20 Gy X-rays,and Transwell was used to detect the migration ability of the three types of cells after irradiations.The co-culture systems of HPMSCs/Leptin or HPMSCs/NC with HGnF,HOK and HUVEC were constructed,and CCK-8 was used to detect the proliferative activity of the three types of cells at 24 h,48 h,72 h,and 96 hafter irradiation,and ELISA to detect the levels of expressed inflammatory factors(human IL-1α,IL-1β,IL-6,IL-8,GM-CSF and TNF-α)by the three types of cells after irradiation at 24 h,48 hand 72 h.Results HPMSCs/Leptin promoted the migration of HUVEC and inhibited the migration of HGnF and HOK.HPMSCs/Leptin could enhance the proliferation ability of HOK and HUVEC after irradiation injury,but the effect on the proliferation of HGnF is not obvious.HPMSCs/Leptin could promote the secretion of human IL-1α,IL-6,GM-CSF and TNF-αby irradiated HGnF,HOK and HUVEC,but it had no obvious effect on the secretion of human IL-1βand IL-8.Conclusion HPMSCs/leptin could promote the migration of irradiated HUVECs.HPMSCs/leptin could promote cell proliferation of irradiated HUVECs、HOKs.HPMSCs/leptin increased secretion of pro-inflammatory cytokines(human GM-CSF,IL-la,and IL-6)in the co-culture system of HPMSCs/leptin and irradiated HUVECs、GnFs、HOKs.Conclusion:Human placental-derived mesenchymal stem cells transfected with leptin(HPMSCs/Leptin)could play apositive role in promoting the healing process of combined radiation and wound injury by supplementing the number of oral mucosal repair cells,regulating inflammation,and

关 键 词：瘦素 人胎盘源间充质干细胞 口腔颌面部创伤 放创复合伤 

分 类 号：R739.8[医药卫生—肿瘤]