石菖蒲挥发油促进牙周炎牙周膜干细胞增殖及迁移的作用机制研究  

The mechanism of Acorus tatarinowii volatile oil promoting proliferation and migration of periodontal ligament stem cells in periodontitis

作　　者：王婧 李召宝 李艳霞[2] WANG Jing;LI Zhaobao;LI Yanxia(Clinics of Stomatology,Cangzhou Central Hospital,Cangzhou 061000,China;Department of Stomatology,Cangzhou Integrated Traditional Chinese and Western Medicine Hospital,Cangzhou 061000,China)

机构地区：[1]沧州市中心医院口腔门诊,河北沧州061000 [2]河北省沧州中西医结合医院口腔科,河北沧州061000

出　　处：《长春中医药大学学报》2023年第4期400-406,共7页Journal of Changchun University of Chinese Medicine

基　　金：河北省2021年度医学科学研究课题计划(20210670);2021年沧州市科技计划自筹经费项目(213106010)。

摘　　要：目的研究石菖蒲挥发油调控磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路对脂多糖(LPS)诱导的人牙周膜干细胞增殖及迁移的影响。方法体外培养人牙周膜干细胞,分为对照组(不做干预),LPS组(8μg·mL^(-1)LPS),实验组[在8μg·mL^(-1)LPS的基础上加入50、100、150 mg·L^(-1)浓度的石菖蒲挥发油,干预24 h,细胞计数试剂盒-8(CCK-8)测定细胞活力并筛选确定150 mg·L^(-1)石菖蒲挥发油为最适浓度]。再将细胞分为对照组(不做干预),LPS组(8μg·mL^(-1)LPS),实验组(8μg·mL^(-1)LPS+150 mg·L^(-1)石菖蒲挥发油),LPS+抑制剂组(8μg·mL^(-1)LPS+10μmol·L^(-1)PI3K/AKT通路抑制剂LY294002),石菖蒲挥发油+LPS+抑制剂组(8μg·mL^(-1)LPS+150 mg·L^(-1)石菖蒲挥发油+10μmol·L^(-1)PI3K/AKT通路抑制剂LY294002),石菖蒲挥发油+LPS+激动剂组(8μg·mL^(-1)LPS+150 mg·L^(-1)石菖蒲挥发油+10μmol·L^(-1)PI3K/AKT通路激动剂SC79),干预24 h。用酶联免疫吸附试验(ELISA)检测炎症因子白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的表达水平;采用CCK-8法测定人牙周膜干细胞的细胞活力;采用5-乙炔基-2’脱氧尿嘧啶核苷(EdU)细胞增殖实验测定人牙周膜干细胞的增殖率;采用细胞划痕实验测定牙周膜干细胞的迁移率;采用蛋白免疫印迹法(WB)检测人牙周膜干细胞周期素D1(Cyclin D1)及PI3K/AKT通路相关蛋白表达水平。结果与对照组比较,LPS组细胞炎症因子IL-6、IL-1β和TNF-α表达水平显著升高,细胞活力显著降低(P<0.05),与LPS组比较,实验组加入不同浓度的石菖蒲挥发油后细胞活力提升,石菖蒲挥发油浓度为150 mg·L^(-1)时细胞活力显著提高并且炎症因子显著降低(P<0.05),所以确定150 mg·L^(-1)石菖蒲挥发油是最佳浓度,进行进一步的实验。与对照组比较,LPS组细胞增殖率和Cyclin D1蛋白表达水平显著降低,p-PI3K/PI3K和p-AKT/AKT蛋白表达水平显著升高(P<0.05);与LPS组比较,�Objective To study the effect of Acorus tatarinowii volatile oil on the proliferation and migration of human periodontal ligament stem cells(PDLSCs)induced by LPS by regulating phosphatidylinositol 3 kinase(PI3K)/the protein kinase B(AKT)signaling pathway.Methods Human periodontal ligament stem cells were cultured in vitro and divided into the control group(no intervention),LPS group(8μg·mL^(-1) lipopolysaccharide),and experimental group[adding 50,100 and 150 mg·L^(-1) Acorus tatarinowii volatile oil on the basis of 8μg·mL^(-1) lipopolysaccharide,intervening for 24 hours,cell counting kit-8(CCK-8)was used to determine cell viability and the optimum concentration of volatile oil was 150 mg·L^(-1)].Cells were then divided into the control group(without intervention)and LPS group(8μg·mL^(-1) LPS),experimental group(8μg·mL^(-1) LPS+150 mg·L^(-1) Acorus tatarinowii volatile oil),experimental group,LPS+inhibitor group(8μg·mL^(-1) lipopolysaccharide+10μmol·L^(-1) PI3K/AKT pathway inhibitor LY294002),Acorus tatarinowii volatile oil+LPS+inhibitor group(8μg·mL^(-1) lipopolysaccharide+150 mg·L^(-1) Acorus tatarinowii volatile oil+10μmol·L^(-1) PI3K/AKT pathway inhibitor LY294002),Acorus tatarinowii volatile oil+LPS+agonist group(8μg·mL^(-1) lipopolysaccharide+150 mg·L^(-1) Acorus tatarinowii volatile oil+10μmol·L^(-1) PI3K/AKT pathway agonist SC79),intervening for 24 hours.The expression levels of interleukin-6(IL-6),IL-1βand tumor necrosis factor-α(TNF-α)were detected by enzyme linked immunosorbent assay(ELISA).The viability of human periodontal ligament stem cells was measured by CCK-8 method.The proliferation rate of human periodontal ligament stem cells was measured by 5-ethynyl-2'deoxyuridine(EdU)cell proliferation assay.Cell scratch test was used to determine the migration rate of periodontal ligament stem cells.The expression levels of Cyclin D1 and PI3K/AKT pathway related proteins in human periodontal ligament stem cells were detected by western blotting(WB).Results Compared with the con

关 键 词：牙周炎 牙周膜干细胞 石菖蒲挥发油 脂酰肌醇3激酶/蛋白激酶B信号通路 增殖 迁移 

分 类 号：R781.42[医药卫生—口腔医学]