基于基因组学比较左归丸和戊酸雌二醇防治绝经后骨质疏松症的作用机制

目的 采用生物信息学分析技术深入挖掘骨髓间充质干细胞（BMSCs）基因表达谱改变，比较左归丸和其阳性对照药戊酸雌二醇防治去卵巢所致骨质疏松症的关键基因与生物学通路。方法 36只大鼠分为模型组、假手术组、左归丸组和戊酸雌二醇组，后2组术后2周开始相应予左归丸和戊酸雌二醇灌胃治疗至第4、8、12周，其余2组蒸馏水灌胃。取材并培养、纯化BMSCs后制备基因表达谱芯片。采用差异基因鉴定、生物学通路富集与相关性分析、WGCNA分析及PPI网络分析等多种生物信息学方法逐层深入进行分析。结果 术后12周，模型组大鼠各位点骨密度降低，而左归丸组和戊酸雌二醇组大鼠骨密度均高于模型组，提示造模成功，中西药均可提高去势大鼠骨密度。差异分析显示相比模型组，左归丸组在术后4、8、12周分别筛选了405、403和618个差异基因；戊酸雌二醇组筛选了78、326和232个差异基因。通路分析发现许多差异基因与炎症和免疫反应通路高度相关，如Il1b、Tnf与病毒感染有关，Itga2、Fgf21同PI3K-Akt信号通路有关，Cxcl12同白细胞经内皮迁移相关等。WGCNA分析结果也显示某些重要节点与PI3K-Akt信号通路及炎症性肠病有关。而PPI网络分析中左归丸组关联到了338个节点，戊酸雌二醇组则包括133个。结论 在骨髓微环境中，左归丸和戊酸雌二醇都具有调节炎症和免疫反应的作用，而本研究所筛选出的通路及基因有可能作为其防治本病的潜在靶标。 

Genomic Comparison of Mechanisms Underlying Zuogui Pill and Estradiol Valerate in Prevention and Treatment of Postmenopausal Osteoporosis

OBJECTIVE To compare the potential genes and pathways involved in the preventive treatment of ovariectomized (OVX)-induced osteoporosis in bone marrow mesenchymal stem cells (BMSCs) obtained from OVX rats treated with Zuogui Pill and Estradiol valerate. METHODS A total of 36 rats were divided into four groups: Model， Sham (Control)， Zuogui Pill， and Estradiol valerate. The rats in the last two groups were intragastrically administered with Zuogui Pill or Estradiol valerate， respectively， from 2 weeks post operation to 4， 8， and 12 weeks post operation. The rats in the other two groups were treated with distilled water. The gene expression patterns in BMSCs were investigated， which were subjected to bioinformatics analyses， including identification of differentially expressed genes (DEGs)， pathway enrichment and correlation analyses， weighted correlation network analysis (WGCNA)， and protein-protein interaction (PPI) network construction. RESULTS At 12 weeks post operation， bone mineral density in OVX model group significantly decreased. Meanwhile， bone mineral densities in Zuogui Pill and Estradiol valerate groups were significantly higher than that in the model group， indicating that the models were constructed successfully， and both traditional Chinese medicine and western treatment could improve the bone density of OVX rats. Numerous DEGs were identified in different groups at different time points. Difference analysis showed that compared with the model group， 405， 403 and 618 DEGs were screened in Zuogui Pill treatment group at 4， 8 and 12 weeks after surgery， respectively， and 78， 326 and 232 DEGs in Estradiol valerate group. Pathway analysis showed that these DEGs were closely related to inflammation- and immunity-associated pathways， such as herpes simplex infection (Il1b and Tnf)， the PI3K-Akt signaling pathway (Itga2 and Fgf21)， and leukocyte transendothelial migration (Cxcl12). WGCNA identified several key modules that were significantly linked to the PI3K-Akt signaling pathway and inflammatory bowel disease. The PPI network in the Zuogui Pill group included 388 nodes and 133 nodes in the Estradiol valerate group. CONCLUSION The results of this study suggest the importance of inflammation and immunity in the pathogenesis of OVX-induced osteoporosis in bone marrow microenvironment. The identified pathways and genes may serve as therapeutic targets for this disease.