基于网络药理学探讨五子衍宗丸对环磷酰胺所致骨髓抑制的治疗作用机制OA

Objective:To investigate the therapeutic effects and mechanisms of the Wuzi Yanzong Pill(五子衍宗丸,WYP)in the treatment of cyclophosphamide(CTX)-induced myelosuppression.Methods:Based on the web-based pharmacological search,87 active components of WYP and 49 key targets for intervening in myelosuppressive process were screened.AKT1,MAPK1,EGFR were screened as the key target genes based on the KEGG enrichment analysis and degree sorting.A mouse model of CTX-induced myelosuppression was constructed,forty mice were randomly divided into 5 groups,the control group,the model group(CTX),the low dose WYP group(WYP-L),the medium dose WYP group(WYP-M),and high dose WYP group(WYP-H).Blood cell count,bone marrow nucleated cell count,bone marrow cell proliferation,cycle and apoptosis were detected.Hematopoietic stem cells(HSCs)were detected in the femur by hematoxylin-eosin staining(HE staining),and the damage of the bone marrow tissue was evaluated by HE staining and scanning electron microscopy.enzyme-linked immunosorbent assay(ELISA)was used to detect changes in the expression levels of serum inflammatory factors IL-1β,IL-6 and TNF-α,haematopoietic-related factors SCF,EPO,TPO and GM-CSF.qRT-PCR was used to detect the changes in the expression levels of c-kit and GM-CSF in bone marrow tissues,EPO and TPO in renal tissues.The protein expression levels of AKT1,MAPK1 and EGFR in bone marrow tissues were examined to evaluate the efficacy and regulatory mechanism of WYP on CTX-induced myelosuppression.Results:Compared with the control group,CTX-induced mice in the model group lost weight and showed significant decreases in PLT,HGB,RBC,WBC.The bone marrow structure was damaged,with a dramatic reduction of nucleated cells in the lumen,disturbed cell arrangement,increased number of fat cells,and multiple haemorrhages.Bone marrow cells in G1 and G2 phases were increased,and cells in S phase were decreased.Meanwhile,CTX caused a decrease in proliferation and a large number of apoptotic cells in the bone marrow of mice.Serum levels of IL-6,TNF-α and IFN-γ inflammatory factors were significantly increased,and the levels of haematopoietic-related factors SCF,EPO,TPO and GM-CSF were significantly decreased.The mRNA expression levels of c-kit,GM-CSF,AKT1,MAPK1 and EGFR in bone marrow tissues,and EPO and TPO in kidney tissues were significantly decreased.After gavage of different doses of WYP,the above indexes were reversed to different degrees in a good dose-dependent manner,with WYP-H being the best.Conclusion:WYP is able to ameliorate CTX-induced haematological abnormalities,bone marrow tissue damage,inflammation and haematopoietic insufficiency through the AKT1/MAPK1/EGFR pathway,thereby alleviating myelosuppression.