复方生脉成骨胶囊调控miR-135b-5p对小鼠激素性股骨头坏死及骨代谢的影响OA

Objective:To study the improvement effect and mechanism of compound Shengmai Chenggu capsules on steroid-induced femoral head necrosis in mice based on the Bushen Huoxue theory.Methods:Mice were randomly divided into the normal group and the model group.The steroid-induced femoral head necrosis model was established using lipopo-lysaccharide combined with methylprednisolone.After the modeling was completed,they were randomly divided into the model group,the compound Shengmai Chenggu capsules group,the Agomir group,the compound Shengmai Chenggu cap-sules+Agomir group,and the alendronate sodium group,with 12 rats in each group.In the compound Shengmai Chenggu capsules group,mice received compound Shengmai Chenggu capsules(5.46 g/kg)by gavage once a day.In the Agomir group,miR-135b-5p Agomir(14 nmol/L)was injected into the tail vein at a volume of 200 μL per mouse once for 2 weeks.In the compound Shengmai Chenggu capsules+Agomir group,compound Shengmai Chenggu capsules(5.46 g/kg)were administered intragastrically once a day,and miR-135b-5p Agomir(14 nmol/L)was injected into the tail vein once for 2 weeks.In the alendronate sodium group,alendronate sodium(1.3 mg/kg)was administered intragastrically once a day.The normal group and the model group were administered an equivalent volume of distilled water by gavage,all groups were continuously treated for 8 weeks.The level of miR-135b-5p was determined by real-time quantitative poly-merase chain reaction(qPCR),the morphology of the femoral head in mice was examined using a micro-computed tomo-graphy scanner(Micro-CT),and the number of trabeculae,the thickness of trabeculae and the separation of trabeculae were calculated.The levels of osteocalcin,alkaline phosphatase(ALP),type Ⅰ collagen amino-terminal propeptide(PINP),tartrate-resistant acid phosphatase(TRAP),parathyroid hormone(PTH),and bone alkaline phosphatase(BALP)in mouse serum were determined using enzyme-linked immunosorbent assay(ELISA)kits.The levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),superoxide dismutase(SOD),malondialdehyde(MDA),and lipid perox-ide(LPO)in mouse serum were determined using an automatic biochemical analyzer.The histopathological changes in the femoral head of mice were examined by hematoxylin-eosin(HE)staining.The mRNA and protein levels of Runt-related transcription factor 2(Runx2),bone morphogenetic protein 2(BMP-2),and osteogenic transcription factor antibody(Osterix)in the femoral head of mice were determined by real-time quantitative PCR and Western Blot,respectively.Results:Compared with the normal group,the levels of miR-135b-5p,trabecular bone separation,serum ALP,TRAP,PTH,TNF-α,IL-6,MDA and LPO in the model group were increased(P<0.05);the number of trabeculae in the femoral head,trabecular thickness,the levels of serum osteocalcin,PINP,BALP and SOD,as well as the mRNA and protein levels of Runx2,BMP-2 and Osterix were decreased(P<0.05).Compared with the model group,the levels of miR-135b-5p,trabecular bone separation,serum ALP,TRAP,PTH,TNF-α,IL-6,MDA,and LPO in both compound Shengmai Chenggu capsules group and the alendronate sodium group were decreased(P<0.05),the number of trabeculae of the femoral head,the thickness of trabeculae,the levels of serum osteocalcin,PINP,BALP and SOD,the mRNA and protein levels of Runx2,BMP-2 and Osterix increased(P<0.05),and the histopathological structure of the femoral head tissue was sig-nificantly improved.Compared with the normal group,the model group and compound Shengmai Chenggu capsules group,the levels of miR-135b-5p,trabecular bone separation,serum ALP,TRAP,PTH,TNF-α,IL-6,MDA and LPO in the Ag-omir group were increased(P<0.05).The number of trabeculae of the femoral head,the thickness of trabeculae,the lev-els of serum osteocalcin,PINP,BALP and SOD,as well as the mRNA and protein levels of Runx2,BMP-2 and Osterix de-creased(P<0.05).Compared with the Agomir group,the levels of miR-135b-5p,trabecular bone separation,serum ALP,TRAP,PTH,TNF-α,IL-6,MDA and LPO in the compound Shengmai Chenggu capsules+Agomir group were de-creased(P<0.05),the number of trabeculae of the femoral head,the thickness of trabeculae,the levels of serum osteocal-cin,PINP,BALP and SOD,as well as the mRNA and protein levels of Runx2,BMP-2 and Osterix increased(P<0.05).Conclusion:The compound Shengmai Chenggu capsules can significantly inhibit the destruction of femoral head tissue structure in mice with steroid-induced femoral head necrosis,promote the pathological recovery of the femoral head,regu-late bone metabolism,inhibit systemic inflammatory responses and oxidative stress injury in mice,and promote osteoblast differentiation in the femoral head.The mechanism may be related to the regulation of miR-135b-5p.