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作 者:王婷 杜令倩[2] WANG Ting;DU Lingqian(Department of General Dentistry,School and Hospital of Stomatology,Cheeloo College of Medicine,Shandong University&Shandong Key Laboratory of Oral Tissue Regeneration&Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration,Jinan 250012,China)
机构地区:[1]山东大学齐鲁医学院口腔医学院·口腔医院综合科山东省口腔组织再生重点实验室山东省口腔生物材料与组织再生工程实验室,山东济南250012 [2]山东大学第二医院口腔科,山东济南250033
出 处:《口腔医学》2022年第8期681-687,共7页Stomatology
基 金:山东省自然科学基金青年项目(ZR2020QH159)。
摘 要:目的 观察生长激素释放肽(Ghrelin)对牙周膜干细胞(periodontal ligament stem cells,PDLSCs)增殖、凋亡的影响,探究其可能机制。方法 通过流式细胞术对培养的PDLSCs进行表面抗原鉴定,采用茜素红及油红O染色鉴定PDLSCs多向分化能力。CCK-8检测Ghrelin对PDLSCs增殖效应的影响;活死细胞染色检测Ghrelin作用下细胞状态;流式细胞术AnnexinV-FITC/PI双染法、TUNEL染色分别检测Ghrelin及其受体GHS-R1α拮抗剂D-Lys3-GHRP-6(DLS)对饥饿诱导下PDLSCs凋亡的作用。结果 本研究中所培养的PDLSCs为间充质来源的多能干细胞。100、200 ng/mL Ghrelin能够显著促进PDLSCs的增殖(P<0.05)。Ghrelin可抑制饥饿诱导的细胞死亡及凋亡,且凋亡抑制效应可以被DLS所逆转。结论 Ghrelin促进PDLSCs增殖,抑制无血清饥饿诱导的细胞凋亡,可能与Ghrelin激活PDLSCs细胞表面受体GHS-R1α有关。Objective To observe the effects of Ghrelin on proliferation and apoptosis of periodontal ligament stem cells(PDLSCs) and explore its possible mechanism.Methods Surface antigens of cultured PDLSCs were identified by flow cytometry,and the multi-directional differentiation ability of PDLSCs was identified by alizarin red staining and oil red O staining.The effect of Ghrelin on the proliferation of PDLSCs was detected by cell counting kit-8(CCK-8) assay.The effect of Ghrelin on cell state of PDLSCs was detected by live/dead cell staining.Apoptotic effects of Ghrelin and its receptor GHS-R1α antagonist D-Lys3-GHRP-6(DLS) on starvation-induced PDLSCs were detected by flow cytometry AnnexinV-FITC/PI double staining and TUNEL staining,respectively.Results Cells cultured in this study were mesenchymal-derived pluripotent stem cells.Moreover,100 and 200 ng/mL Ghrelin significantly promoted the proliferation of PDLSCs(P<0.05).Moreover,Ghrelin inhibited starvation-induced cell death and apoptosis,and this anti-apoptotic effect was reversed by DLS.Conclusion Ghrelin promotes the proliferation of PDLSCs and inhibits serum-free starvation-induced apoptosis.This process might be related to the binding of Ghrelin to PDLSCs surface receptor GHS-R1α.
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