中医正骨2025，Vol.37Issue(5)：6-20,29,16.
补肾活血胶囊防治大鼠激素性股骨头坏死的效果及其微生物组学和代谢组学作用机制研究
Efficacy of Bushen Huoxue Jiaonang(补肾活血胶囊)against steroid-induced osteonecrosis of the femoral head in rats and its microbiomics and metabonomics mechanisms:an experimental study
摘要
Abstract
Objective:To observe the outcomes of Bushen Huoxue Jiaonang(补肾活血胶囊,BSHXJN)against steroid-induced osteo-necrosis of the femoral head(SONFH)in rats,and to analyze and explore its underlying mechanisms based on microbiomics and metabono-mics.Methods:Twenty-four rats were randomized into blank group,model group,and BSHXJN group,with 8 ones in each group.All rats but the ones in blank group were modeled by intramuscular injection of methylprednisolone sodium succinate into the right gluteal region for inducing SONFH on the first three days of each week,once a day for consecutive 3 weeks.During the SONFH modeling,the rats in BSHXJN group were intervened by intragastric administration with BSHXJN solution at a crude dose of 5.4 g/kg/day,while the ones in blank group and model group with the equivalent volume of purified water,once a day for consecutive 6 weeks.After the end of the last intervention,the rats were sacrificed and their bilateral femur heads and faeces were harvested.The right femur heads were scanned by a Micro-CT scanner to observe the imaging changes,and analyze the trabecular bone volume(TBV)and bone volume(BV).After that,the left femur heads were stained with HE to observe the changes in trabecular quantity and the degree of adipocyte infiltration.Furthermore,the protein expression levels of osteogenic markers including Runt-related transcription factor 2(RUNX2)and Osterix in the rat right femur heads were detected by using Western Blot,the microbial communities in the rat fecal samples were identified by 16S ribosomal DNA(rDNA)sequencing,followed by differential gut microbiota analysis and functional prediction.Moreover,the metabolites in the fecal samples of rats in each group were detected by liquid chromatography-mass spectrometry(LC-MS),and the differential gut metabolite analysis and KEGG pathway enrichment analysis were performed.Results:①Micro-CT analysis on femur heads.The femur heads in rats of control group exhibited regular contours with well-organized subchondral trabeculae,and no cystic changes,whereas the femur heads in model group displayed irregular contours,ac-companied by missed and disordered subchondral trabeculae,and some cystic structures,while these deteriorations were mitigated in BSHXJN group compared to model group.Apart from that,the TBV and BV were smaller in model group compared to blank group,and were greater in BSHXJN group compared to model group(P=0.002,P=0.000;P=0.002,P=0.000),with no significant difference between BSHXJN group and blank group(P=0.994,P=0.186).②The pathological findings in femur heads.In blank group,the subchondral tra-beculae of femur heads presented uniform distribution with no adipocyte infiltration;while that in the model group demonstrated sparse or discontinuous distribution,accompanied by adipocyte infiltration;whereas,the changes were ameliorated in BSHXJN group compared to model group,manifesting as denser subchondral trabeculae and significantly mitigated adipocyte infiltration in the femur heads.③The pro-tein expression levels of osteogenic markers.The relative protein expression levels of RUNX2 and Osterix in the femur heads were lower in model group compared to blank group,and were higher in BSHXJN group compared to model group(P=0.000,P=0.001;P=0.000,P=0.000),with no significant difference between BSHXJN group and blank group(P=0.279,P=0.509).④The gut microbiota.At the phy-lum level,the abundance of Patescibacteria,Verrucomicrobiota,and Fusobacteriota in the gut microbiota decreased in BSHXJN group com-pared to model group;At the genus level,the abundance of Psychrobacter in the gut microbiota increased,while that of Candidatus Sacchari-monas,Turicibacter and Akkermansia decreased in BSHXJN group compared to model group;Furthermore,the gene abundance related to flavonoid biosynthesis,isoflavonoid biosynthesis,and hypertrophic cardiomyopathy(HCM)in the gut microbiota was significantly lower in BSHXJN group compared to model group.⑤The gut metabolites.The differential gut metabolites between BSHXJN group and model group were sorted by the variable importance in projection(VIP)from high to low,and the top 20 ones were Cholesterol Sulfate,Oleamide,Ephed-rine,trans-Vaccenic Acid,Hypoxanthine,Erythraline,N-Acetylneuraminic Acid(Neu5Ac),2-Piperidone,L-Carnitine,4-Pyridoxic Acid,Phenanthren-6-one-containing Compound,Cortisone,Butyric Acid,Deoxyadenosine,Palmitamide,Protopine,Erucamide,Androstenedione,trans-Dehydroandrosterone,and N-Methyl-1-glutamic Acid,and they were mainly enriched in Prostate Cancer pathways(KEGG:ko05215),Steroid hormone biosynthesis pathways(KEGG:ko00140),Alpha-linolenic acid metabolism pathways(KEGG:ko00592),Vitamin B6 me-tabolism pathways(KEGG:ko00750),and pathways in cancer(KEGG:ko05200).Conclusion:BSHXJN can ameliorate the bone micro-structure of femur heads,promote osteogenesis,and inhibit osteonecrosis.It may work by modulating the gut microbiota and gut metabolite.关键词
股骨头坏死/激素类/补肾活血胶囊/胃肠道微生物群/代谢物/代谢产物学/大鼠Key words
femur head necrosis/hormones/Bushen Huoxue Jiaonang/gastrointestinal microbiome/metabolites/metabolomics/rats引用本文复制引用
李威,张博淳,周忠起,李广政,李刚,梁学振..补肾活血胶囊防治大鼠激素性股骨头坏死的效果及其微生物组学和代谢组学作用机制研究[J].中医正骨,2025,37(5):6-20,29,16.基金项目
国家自然科学基金项目(82205154,82074453) （82205154,82074453）
山东省自然科学基金项目(ZR2024MH156) （ZR2024MH156）
