Effects of elderberry on bone protection,m6A methylation and osteogenic differentiation of bone marrow mesenchymal stem cells in alcoholic bone remodeling
Objective:To investigate the effects of elderberry on bone protection,m6A methylation and osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs)after alcoholic bone remodeling(ABR).Methods:A total of 36 male SD rats were randomly divided into blank group,model group,elderberry low-dose group and elderberry high-dose group,9 rats in each group.Rats in model group,elderberry low-dose group and elderberry high-dose group were given intraperitoneal injection of 20％ethanol once a day for 12 weeks.The blank group was intraperitoneally injected with the same amount of normal saline.On the first day of modeling,elderberry low-dose and high-dose group was given 340,680 mg/kg elderberry on the basis of model group,blank group and model group were given the same amount of normal saline intragastric administration,once a day.The changes of bone trabecula and collagen fibers were detected by HE and Masson staining.The expression of bone metabolic marker genes Runx2,COL1A2,BMP2 and OPG were detected by qRT-PCR.The methylation level of Runx2 m6A was detected by methylated RIP-qPCR.Runx2 protein levels were detected by Western Blot.The content of reactive oxygen species was detected by DCFH-DA staining.Alizarin red staining was used to detect the formation of calcified nodules.Results:Compared with model group,the number of bone trabeculae,collagen fiber content increased,elastic modulus,elastic load and maximum stress significantly increased(P＜0.05),Runx2 methylation and protein levels significantly increased(P＜0.05),while the content of reactive oxygen species was decreased,all of which were close to the level of blank group,bone formation related genes BMP2,Runx2,COL1A2,OPG mRNA expression significantly increased(P＜0.05)in elderberry low-dose group and elderberry high-dose group.Conclusion:By regulating m6A RNA methylation modification and osteogenic differentiation of BMSCs,elderberry can increase the number of bone trabeculae and the content of collagen fibers to restore bone homeostasis,thus antagonizing the occurrence and development of ABR.
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