湖南中医药大学学报2025，Vol.45Issue(5)：818-824,7.DOI:10.3969/j.issn.1674-070X.2025.05.006
基于有氧糖酵解探究补骨生髓方含药血清通过介导HIF-1α促进MC3T3-E1细胞成骨分化的作用
Promotion of osteogenic differentiation of MC3T3-E1 cells by serum containing Bugu Shengsui Decoction through mediating HIF-1α based on aerobic glycolysis
摘要
Abstract
Objective To investigate the effects of the Bugu Shengsui Deocoction(BGSSF)containing serum on osteogenic differentiation of MC3T3-E1 cells by mediating aerobic glycolysis through hypoxia-inducible factor-1α(HIF-1α).Methods A total of 18 SD rats were randomly divided into a drug-containing serum group and a blank serum group,with nine rats in each group.The drug-containing serum group was administered BGSSD solution by gavage at a raw drug dose of 5.87 g/(kg·d),while the blank serum group was given distilled water by gavage at a volume of 20 mL/(kg·d).After nine consecutive gavages,blood was collected from the abdominal aorta to obtain serum containing BGSSD and blank serum.MC3T3-E1 cells were divided into a blank serum group,a drug-containing serum group,and an aerobic glycolysis inhibitor group.Additionally,MC3T3-E1 cells stably transfected with HIF-1α lentivirus were designated as the SiRNA HIF-1α group.The blank serum group was cultured in α-MEM medium containing 20％blank serum,while both the drug-containing serum group and the SiRNA HIF-1α group were cultured in α-MEM medium containing 20％serum containing BGSSD.The aerobic glycolysis inhibitor group was cultured in α-MEM medium containing 20％BGSSF containing serum and 8 mmol/L of the inhibitor 2-deoxyglucose.Alkaline phosphatase(ALP)staining was used to observe the black precipitates in the four groups after seven days of culture.ELISA was employed to determine the content of ALP,pyruvic acid(PA),and lactic acid(LA)in the cell supernatant after 48 hours of culture.qRT-PCR and Western blot were utilized to measure the relative mRNA expression levels and protein expression amounts of HIF-1α,Runt-related transcription factor 2(RUNX2),and pyruvate dehydrogenase kinase 1(PDK1)in the cells after 48 hours of culture.Results Compared with the blank serum group,the drug-containing serum group exhibited a notable increase in black precipitates with deeper coloration,along with elevated levels of ALP,LA,and PA(P＜0.05).Additionally,the relative mRNA expression levels and protein expression amounts of PDK1,RUNX2,and HIF-1α all increased in the drug-containing serum group(P＜0.05).Compared with the drug-containing serum group,the SiRNA HIF-1α group and the aerobic glycolysis inhibitor group showed reduced black precipitates with lighter coloration,as well as decreased levels of ALP,LA,and PA(P＜0.05).The relative mRNA expression levels and protein expression amounts of PDK1 and RUNX2 also decreased in these two groups(P＜0.05).The relative mRNA expression levels and protein expression amounts of HIF-1α reduced in the SiRNA HIF-1α group(P＜0.05).Conclusion BGSSD may promote osteogenic differentiation of MC3T3-E1 cells by mediating aerobic glycolysis through HIF-1α,and its mechanism of action might be related to the regulation of the HIF-1α/PDK1 pathway.关键词
骨质疏松症/补骨生髓方/有氧糖酵解/MC3T3-E1细胞/骨形成Key words
osteoporosis/Bugu Shengsui Decoction/aerobic glycolysis/MC3T3-E1 cells/bone formation分类
中医学引用本文复制引用
尹煜辉,李琰,齐保玉,刘平,朱立国,刘宁,魏戌..基于有氧糖酵解探究补骨生髓方含药血清通过介导HIF-1α促进MC3T3-E1细胞成骨分化的作用[J].湖南中医药大学学报,2025,45(5):818-824,7.基金项目
国家自然科学基金面上项目(82174416) （82174416）
中国中医科学院基本科研业务费新入职青年科研人员培养专项课题(ZZ17-XRZ-059) （ZZ17-XRZ-059）
中国中医科学院望京医院自主选题专项课题(WJYY-ZZXT-2023-16). （WJYY-ZZXT-2023-16）
