加味七方胃痛颗粒通过重塑肿瘤相关巨噬细胞极化状态抗胃癌的作用机制研究OA

Objective To investigate the regulatory role and mechanism of Modified Qifang Weitong Granule(加味七方胃痛颗粒,MQWG)on tumor-associated macrophages polarization in HGC-27 gastric cancer-bearing nude mice.Methods A subcutaneous xeno-graft model of HGC-27 gastric cancer was established in nude mice.The antitumor effect of MQWG was evaluated based on relative tumor proliferation rate and hematoxylin-eosin(HE)staining.The key signaling pathways of MQWG against gastric cancer were screened using network pharmacology.Serum levels of TNF-α and IL-10 were measured by ELISA.Immunofluorescence was performed to detect the expression of macrophage markers(CD86,CD206)and immune cells(CD4+T,CD8+T)in tumor tissues.RT-qPCR was used to assess the mRNA expression of iNOS,Arg-1,IL-10 and IL-1βin tumor tissues.Protein immunoblotting to verify whether the PI3K-AKT pathway screened by network pharmacology is a key mediator of the anti-gastric cancer pathway of MQWG.Results Com-pared with the control group,the tumor volume in the Chinese medicine groups was significantly reduced,accompanied by increased tumor cell necrosis and degeneration,as well as a marked decrease in relative tumor proliferation rate.Network pharmacology identified the PI3K-AKT signaling pathway as the key signaling pathway of MQWG against gastric cancer.Serum IL-10 levels were decreased,while TNF-α levels were elevated.Tumor tissues exhibited increased CD86 expression and decreased CD206 expression.Additionally,mRNA expression of Arg-1and IL-10 was downregulated,whereas iNOS and IL-1βmRNA expression was upregulated.Validation of the PI3K-AKT signaling pathway by Western blot(WB)assay revealed a significant decrease in the p-PI3K/PI3K and pAKT/AKT ratios,and a decrease in the phosphorylation levels of PI3K and AKT proteins.Conclusion MQWG promotes M1-type polarization of TAMs while inhibiting M2-type polarization,improves the tumor microenvironment.The underlying mechanism may be associated with the suppression of PI3K-AKT signaling pathway activation.