摘要
目的探讨内质网应激通路在棕榈酸(PA)诱导的人牙周膜干细胞(hPDLSCs)凋亡中的作用机制。方法将培养的hPDLSCs随机分为对照组(不含PA的培养基),PA低、中、高浓度(0.1、0.2、0.4 mmol/L)组以探究PA抑制hPDLSCs增殖的作用机制;将培养的hPDLSCs分为对照组(不含PA的培养基)、PA组(0.4 mmol/L)、PA(0.4 mmol/L)+4-PBA(5μmol/L)组以探究4-PBA对PA所致hPDLSCs凋亡的影响。培养24 h后MTT法检测各组hPDLSCs增殖率,Annexin V-FITC/PI探针观察各组hPDLSCs细胞凋亡情况,免疫印迹法检测内质网应激凋亡相关蛋白表达。结果PA低、中、高浓度组hPDLSCs增殖率明显低于对照组(P<0.05);PA高浓度组细胞核红色荧光和细胞膜绿色荧光较对照组明显增加;PA高浓度组IRE1^(phos)、PERK^(phos)、ATF4、ATF6、CHOP表达明显高于对照组(P<0.05);PA中、高浓度组Cleaved caspase-3表达明显高于对照组(P<0.05);与PA低、中浓度组比较,PA高浓度组IRE1^(phos)、PERK^(phos)、ATF6、ATF4、CHOP、Cleaved caspase-3蛋白表达均明显升高(P<0.05);与PA组比较,PA+4-PBA组PERK^(phos)、CHOP及Cleaved caspase-3表达明显下调(P<0.05)。结论PA可能通过调控内质网应激凋亡通路蛋白表达,促进hPDLSCs凋亡。
Objective To investigate the mechanism of endoplasmic reticulum stress pathway in the apoptosis of human periodontal ligament stem cells(hPDLSCs)induced by palmitic acid(PA).Methods The cultured hPDLSCs were randomly divided into control group(medium without PA),low,medium,and high concentrations of PA(0.1,0.2,and 0.4 mmol/L)groups to explore the mechanism of PA inhibiting hPDLSCs proliferation;the cultured hPDLSCs were divided into control group(medium without PA),PA group(0.4 mmol/L)and PA(0.4 mmol/L)+4-PBA group(5μmol/L)to explore the effect of 4-PBA on the apoptosis of hPDLSCs induced by PA.After 24 h of culture,the proliferation rate of hPDLSCs in each group was detected by MTT assay,the apoptosis of hPDLSCs in each group was observed by Annexin V-FITC/PI probe,and the expression of endoplasmic reticulum stress apoptosic-related proteins was detected by Western blot.Results The proliferation rate of hPDLSCs in low,medium,and high PA groups was significantly lower than that in control group(P<0.05).The nuclear red fluorescence and cell membrane green fluorescence of PA high concentration group were increased significantly compared with cortrol group.The expressions of IRE1phos,PERKphos,ATF4,ATF6,and CHOP in PA high concentration group were significantly higher than those in control group(P<0.05).Cleaved caspase-3 expression in PA medium and high concentration groups was significantly higher than that in control group(P<0.05).Compared with PA low and medium concentrations groups,IRE1phos,PERKphos,ATF6,ATF4,CHOP,and Cleaved caspase-3 protein expressions in PA high concentration group were significantly increased(P<0.05).Compared with PA group,PERKphos,CHOP,and Cleaved caspase-3 expressions were significantly down-regulated in PA+4-PBA group(P<0.05).Conclusion PA may promote the apoptosis of hPDLSCs by regulating the protein expression of endoplasmic reticulum stress apoptosis pathway.
作者
刘涛
李晶
鲍翠玉
LIU Tao;LI Jing;BAO Cuiyu(Department of Stomatology,Affiliated Second Hospital of Hubei University of Science and Technology,Hubei Province,Xianning437100,China;Hubei Province Key Laboratory on Cardiovascular,Cerebrovascular,and Metabolic Disorders,Hubei University of Science and Technology,Hubei Province,Xianning437100,China)
出处
《中国医药导报》
CAS
2022年第4期4-8,共5页
China Medical Herald
基金
国家自然科学基金资助项目(51703055)
湖北省自然科学基金项目(2017CFB699)
湖北省咸宁市市级科技计划项目(咸科技发〔2018〕18号-49)
湖北省咸宁市科技创新专项项目(咸财函〔2019〕146号-2)
湖北科技学院校内科研发展基金项目校内培育科研项目资助(2021-22X16)
湖北科技学院五官医学院专项科研基金项目(2020WG14)。
关键词
内质网应激
人牙周膜干细胞
棕榈酸
细胞增殖
凋亡
Endoplasmic reticulum stress
Human periodontal membrane stem cells
Palmitic acid
Cell proliferation
Apoptosis
