沙参麦冬汤调控JAK2/STAT3信号通路对Lewis肺癌模型小鼠化疗增效机制OA

Objective:To predict the mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer via network pharmacology and validate the prediction results in animal experiments.Methods:The potential mechanism through which Shasheng Maidong Tang enhances the efficacy of chemotherapy for lung cancer was predicted by network pharmacology,liquid chromatography-mass spectrometry(LC-MS),and molecular docking methods.C57/BL6 mice were assigned into normal,model,cisplatin,and Shasheng Maidong Tang+cisplatin groups.In addition to the normal group,the remaining groups were injected subcutaneously with 0.2 mL of 1×107 cells·mL-1 Lewis lung cancer cells to establish the Lewis lung cancer model.The daily gavage dose of Shasheng Maidong Tang was 3.58 g·kg-1,and the concentration of cisplatin intraperitoneally injected on every other day was 2 mg·kg-1.Drugs were administered for 14 d.The changes in the tumor volume and the rate of tumor suppression were monitored,and the tumor histopathological changes were observed by hematoxylin-eosin(HE)staining.Enzyme-linked immunosorbent assay was employed to measure the interleukin(IL)-6 and interferon(IFN)-γ levels in peripheral blood.Real-time PCR was performed to quantify the mRNA levels of Janus kinase 2(JAK2),signal transducer and activator of transcription 1(STAT1),and signal transducer and activator of transcription 3(STAT3)in the tumor tissue of mice.Western blot was employed to determine the protein levels of JAK2,STAT3,B-cell lymphoma-2(Bcl-2),cysteinyl aspartate-specific proteinase-3(Caspase-3),and Pim-1 proto1(PIM1)in the tumor tissue.Immunohistochemistry was employed to detect the expression of Bcl-2 and PIM1 in the tumor tissue.Results:Network pharmacological predictions indicated that Shasheng Maidong Tang might enhance the efficacy of chemotherapy for lung cancer by regulating nitrogen metabolism,AGE-RAGE signaling pathway,cancer pathway,and JAK/STAT signaling pathway.The experimental results demonstrated that tumor volume in the cisplatin group and Shasheng Maidong Tang+cisplatin group was reduced compared with the model group,with statistically distinct differences observed on days 14,17,20 post modeling(P＜0.05).Notably,the Shasheng Maidong Tang+cisplatin therapy further decreased tumor volume compared with the cisplatin group,showing marked reductions on days 17 and 20(P＜0.05),consistent with trends visualized in tumor volume comparison charts.The Shasheng Maidong Tang+cisplatin group exhibited higher tumor inhibition rate than the cisplatin group(P＜0.05).Histopathological analysis via HE staining revealed that the tumors in the model group displayed frequent nuclear mitosis,densely arranged cells,hyperchromatic nuclei,and no necrosis.Cisplatin treatment induced partial necrosis and vacuolization,while the Shasheng Maidong Tang+cisplatin group exhibited extensive necrotic regions,maximal vacuolization,disarranged tumor cells,and minimal mitotic activity.Compared with the model group,the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed elevated level of IFN-γ(P＜0.01)and declined level of IL-6(P＜0.01)in the peripheral blood.Compared with the cisplatin group,the Shasheng Maidong Tang+cisplatin group presented elevated level of IFN-γ(P＜0.01)and lowered level of IL-6(P＜0.01)in the peripheral blood.Compared with the model group,the cisplatin group and the Shasheng Maidong Tang+cisplatin groups showed down-regulated mRNA levels of JAK2 and STAT3(P＜0.01)and up-regulated mRNA level STAT1(P＜0.01).Compared with the cisplatin group,the Shasheng Maidong Tang+cisplatin group presented down-regulated mRNA levels of JAK2 and STAT3(P＜0.01)and up-regulated mRNA level of STAT1(P＜0.01).Compared with the model group,the cisplatin group and the Shasheng Maidong Tang+cisplatin group showed down-regulated protein levels of JAK2(P＜0.01),Bcl-2(P＜0.01),PIM1(P＜0.01),and STAT3(P＜0.05),and up-regulated protein level of Caspase-3(P＜0.01).Compared with the cisplatin group,Shasheng Maidong Tang+cisplatin group presented down-regulated protein levels of JAK2(P＜0.01),Bcl-2(P＜0.01),PIM1(P＜0.01),STAT3(P＜0.05),and up-regulated protein level of Caspase-3(P＜0.01).The Bcl-2 and PIM1 expression results obtained by immunohistochemistry were consistent with those of Western blot.Conclusion:Shasheng Maidong Tang may enhance the efficacy of chemotherapy in the mouse model of Lewis lung cancer by regulating the JAK2/STAT3 signaling pathway.