Clinical Observation Of Qilian Mixture In The Treatment Of Non-small Cell Lung Cancer(Deficiency Of Qi And Phlegm Dampness Type)and Experimental Study On Immune Escape Mechanism Of NSCLC Based On ILT4/B7-H3 Pathway

Objective:1.To observe the clinical effect of Qilian mixture on the survival status,quality of life and immune function of patients with advanced non-small cell lung cancer(Deficiency of qi and phlegm dampness type).2.To study the mechanism of Astragaloside inhibiting immune escape of NSCLC through ILT4-PI3K/Akt-B7-H3 pathway by experiment.Methods:1.80 patients with stage ⅢB-Ⅳnon-small cell lung cancer of deficiency of qi and phlegm dampness type were selected and randomly divided into chemotherapy group and combined chemotherapy group with 40 cases in each group.The patients in the chemotherapy group were treated with AP or DP,while those in the combined chemotherapy group were treated with chemotherapy and Qilian mixture.After 4 cycles of chemotherapy,observe and record the effect of solid tumor,immune function,TCM syndrome score,quality of life score before and after treatment,as well as the adverse reactions of the two groups during treatment.2.A549 cell line of NSCLC was used for the experiment.To determine the IC50 of Astragaloside by MTT drug toxicity test,and to observe the effect of drug concentration and time on cell proliferation by MTT cell proliferation test.The cells were divided into four groups:blank group(without drugs),Astragaloside low-dose group(drug concentration is 1/2 times of IC50),Astragaloside middle-dose group(drug concentration is IC50),Astragaloside high-dose group(drug concentration is 2 times of IC50).After drug intervention,Western blot and qRT-PCR were used to detect the effect of Astragaloside on the expression of ILT4-PI3K/Akt-B7-H3 pathway protein and mRNA.Results:1.(1)During the clinical observation,both groups had cases of exfoliation,38 cases in the chemotherapy group and 37 cases in the combined chemotherapy group were finally compared.Before treatment,there was no significant difference in gender,age,clinical stage and observation indexes between the two groups(P>0.05).(2)After four courses of grouping treatment,the combined chemotherapy group showed better curative effect in all aspects compared with the simple chemotherapy group.Traditional Chinese medicine combined with chemotherapy stabilize the weight of patients better(P<0.05).Although there was no significant difference in the curative effect of solid tumor between the two groups(P>0.05),the objective remission rate and disease control rate in the combined chemotherapy group were higher than those in the simple chemotherapy group.(3)After treatment,CEA,CYFRA21-1 and CA-125 in the combined chemotherapy group were lower than those in the simple chemotherapy group(P<0.05).The combination of traditional Chinese medicine and chemotherapy alleviated the inhibition of chemotherapy on immune function.The number of white blood cells,the proportion of lymphocytes and hemoglobin in the combined chemotherapy group were higher than those in the simple chemotherapy group(P<0.05).The CD4+/CD8+of T lymphocyte subsets in the combined chemotherapy group was higher than that in the simple chemotherapy group(P<0.05).(4)The combination of traditional Chinese medicine and chemotherapy can improve the total score of TCM syndrome,especially in cough and expectoration,chest tightness and shortness of breath,asthenia and self perspiration(P<0.05),while chemotherapy alone can aggravate some symptoms.The combined chemotherapy group was superior to the simple chemotherapy group in KPS and ZPS(P<0.05).PFS in the combined chemotherapy group was longer than that in the simple chemotherapy group(P<0.05).The toxic effects of chemotherapy were less in the combined chemotherapy group(P<0.05).2.MTT cell proliferation experiments showed that Astragaloside can effectively inhibit the proliferation of A549 cells,and its inhibitory effect is time-concentration dependent,so as to obtain the IC50 value of Astragaloside on A549 cells.The IC50 of Astragaloside on A549 cells is 28.415μmol/L.Western blot and qRT-PCR showed that Astragaloside could inhibit ILT4-PI3K/Akt-B7-H3 pathway at protein level and mRNA level,and the medium dose was superior(P<0.05).Conclusion:The effect of Qilian mixture combined with chemotherapy in the treatment of NSCLC is better than that of chemotherapy alone.Qilian mixture combined with chemotherapy can not only improve the immunosuppression of chemotherapy,but also improve the TCM syndrome score,improve the state of life function and physical strength,prolong the disease progression free survival time of patients,and reduce the adverse reactions and side effects of chemotherapy on patients.The results of the experiment showed that Astragaloside can significantly reduce the expression of ILT4,inhibit the PI3K/Akt pathway,and reduce the expression of B7-H3,so as to inhibit the immune escape of NSCLC.