中国中药杂志2025，Vol.50Issue(3)：583-589,7.DOI:10.19540/j.cnki.cjcmm.20240719.403
miR-140-5p/BCL2L1 在 HFOB1.19 凋亡、自噬中的作用及补肾健脾活血方的影响
Role of miR-140-5p/BCL2L1 in apoptosis and autophagy of HFOB1.19 and effect of Bushen Jianpi Huoxue Decoction
摘要
Abstract
Osteoporosis(OP)is a senile bone disease characterized by an imbalance between bone remodeling and bone formation.Targeting pathogenesis of kidney deficiency,spleen deficiency,and blood stasis,Bushen Jianpi Huoxue Decoction has a significant effect on the treatment of OP by tonifying kidney,invigorating spleen,and activating blood circulation.MicroRNA(miRNA)and the anti-apoptotic protein B-cell lymphoma-2-like protein 1(BCL2L1)are closely related to bone cell metabolism.Therefore,in this study,the binding of miR-140-5p to BCL2L1 was detected by dual luciferase assay and polymerase chain reaction(PCR).After silencing or overexpressing miR-140-5p,the apoptosis,autophagy,and osteogenic function of human fetal osteoblast cell line 1.19(HFOB1.19)were observed by flow cytometry and Western blot.Bushen Jianpi Huoxue Decoction-containing serum was prepared by intragastric administration of Bushen Jianpi Huoxue Decoction in rats.Different concentrations of Bushen Jianpi Huoxue Decoction-containing serum were used to treat HFOB1.19 with or without miR-140-5p mimic.The expression of osteogenic proteins in each group was observed,and the role of miR-140-5p/BCL2L1 in apoptosis and autophagy of HFOB1.19 was studied,along with the effect of Bushen Jianpi Huoxue Decoction on these processes.As indicated by the dual luciferase assay,miR-140-5p bound to BCL2L1.Flow cytometry and Western blot showed that miR-140-5p promoted apoptosis and inhibited autophagy in HFOB1.19.After intervention with high,medium,and low doses of Bushen Jianpi Huoxue Decoction-medicated serum,compared with the miR-140-5p NC group,the expression of osteocalcin(OCN),osteopontin(OPN),Runt-related transcription factor 2(RUNX2),and transforming growth factor beta 1(TGF-β1)decreased in the miR-140-5p mimic group,while the expression of bone morphogenetic protein 2(BMP2)showed no significant difference under high-dose intervention.Therefore,miR-140-5p/BCL2Ll can promote apoptosis and inhibit autophagy in HFOB1.19.Bushen Jianpi Huoxue Decoction can affect the osteogenic effect of miR-140-5p through BMP2.关键词
骨质疏松症/补肾健脾活血方/miR-140-5p/BCL2L1/凋亡/自噬Key words
osteoporosis/Bushen Jianpi Huoxue Decoction/miR-140-5p/BCL2L1/apoptosis/autophagy引用本文复制引用
陈桐莹,黄宏兴,万雷,付赛,李小韵,刘树华,杨贻富,杨东升,曾韵杰,李扬博,罗丹..miR-140-5p/BCL2L1 在 HFOB1.19 凋亡、自噬中的作用及补肾健脾活血方的影响[J].中国中药杂志,2025,50(3):583-589,7.基金项目
国家自然科学基金项目(82274551,82174395) （82274551,82174395）
广州中医药大学"双一流"与高水平大学学科协同创新团队重点项目(2021XK21) （2021XK21）
广州中医药大学第三附属医院"瞪羚计划"普通项目(KY6032022006) （KY6032022006）
广东省中医药局科研项目(20221178) （20221178）
