葛根素对高糖诱导成骨细胞增殖、凋亡及CHOP通路的影响研究  

Puerarin Has Effect on Proliferation and Apoptosis of Osteoblast Induced by High Glucose and CHOP Pathway

作　　者：洪荣华[1] 李永华[1] 张富国[1] HONG Ronghua;LI Yonghua;ZHANG Fuguo

机构地区：[1]台州市第一人民医院骨科,浙江台州318020

出　　处：《新中医》2020年第7期8-11,共4页New Chinese Medicine

基　　金：台州市社会发展类一般项目(1701KY45)。

摘　　要：目的:探讨葛根素对高糖诱导成骨细胞增殖、凋亡及CHOP通路的影响。方法:设置MC3T3-E1组,葛根素低、中、高剂量组。MC3T3-E1组常规培养,葛根素低、中、高剂量组分别用浓度为20.0μg/mL、40.0μg/mL、80.0μg/mL的葛根素溶液干预培养。3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐法(MTT)法测定各组MC3T3-E1细胞增殖水平,流式细胞法测定各组MC3T3-E1细胞凋亡水平,逆转录-定量聚合酶链反应(RT-qPCR)法测定各组MC3T3-E1细胞转录激活因子4(ATF4)、C/EBP同源蛋白(CHOP) RNA表达水平,Elisa法测定各组MC3T3-E1细胞ATF4、CHOP、超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)蛋白表达水平。结果:与MC3T3-E1组比较,葛根素低、中、高剂量组细胞凋亡率、MDA蛋白表达水平与ATF4、CHOP RNA及蛋白表达水平均降低(P<0.05);OD值、增殖率与SOD、GSH蛋白表达水平均升高(P<0.05)。与葛根素低剂量组比较,葛根素中、高剂量组凋亡率、MDA蛋白表达水平与ATF4、CHOP RNA及蛋白表达水平均降低(P<0.05);OD值、增殖率与SOD、GSH蛋白表达均升高(P<0.05)。与葛根素中剂量组比较,葛根素高剂量组凋亡率、MDA蛋白表达水平与ATF4、CHOP RNA及蛋白表达水平均降低(P<0.05);OD值、增殖率与SOD、GSH蛋白表达水平均升高(P<0.05)。结论:葛根素能拮抗高糖对MC3T3-E1成骨细胞的增殖抑制作用和凋亡促进作用;其机制与葛根素抑制ATF4、CHOP RNA及蛋白的表达拮抗了高糖环境引起的成骨细胞内质网应激,进而减轻成骨细胞氧化应激反应有关。Objective: To discuss the effect of puerarin on proliferation and apoptosis of osteoblast induced by high glucose and CHOP pathway. Methods:Set the MC3 T3-E1 group,the groups of puerarin in low,middle and high doses. The MC3 T3-E1 group received routine culture, while the groups of puerarin in low, middle and high doses received the interfering culture with puerarin solution in 20.0 μg/mL, 40.0 μg/mL and 80.0 μg/mL respectively. Determined the proliferation level of MC3 T3-E1 cell in each group via 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium bromide(MTT) method. Detected the apoptosis level of MC3 T3-E1 cell in each group via flow cytometry method. Determined the expression level of MC3 T3-E1 cell activated transcription factors 4(ATF4) and C/EBP homologous protein(CHOP) RNA via reverse transcription quantitative polymerase chain reaction(RT-qPCR) method. Assessed the expression level of MC3 T3-E1 cell ATF4, CHOP, superoxide dismutase(SOD), malondialdehyde(MDA) and glutathione(GSH) via Elisa method. Results:Comparing with the MC3 T3-E1 group,the apoptosis rate and the expression level of MDA,the expression levels of ATF4,CHOP RNA and protein in the groups of puerarin in low,middle and high doses were deceased(P<0.05);the value of OD,the proliferation rate and the protein expression of SOD and GSH were increased(P<0.05). Comparing with the low-dose puerarin group,the apoptosis rate and the expression level of MDA,the expression levels of ATF4,CHOP RNA and protein in the groups of puerarin in middle and high doses were decreased(P<0.05);the value of OD,the proliferation rate and the protein expression of SOD and GSH were increased(P<0.05). Comparing with the middle-dose puerarin group, the apoptosis rate,MDA expression level and the expression levels of ATF4,CHOP RNA and protein in the group of puerarin of high dose were decreased(P<0.05);the value of OD,the proliferation rate and the protein expression of SOD and GSH were increased(P<0.05). Conclusion:Puerarin can antagonize the effect of high glucose

关 键 词：2型糖尿病 高糖 葛根素 成骨细胞 增殖 凋亡 CHOP通路 细胞实验 

分 类 号：R285.5[医药卫生—中药学] R587.1[医药卫生—中医学]