Exploration Of The Regulation Mechanism Of Jianpi Huatan Decot Ion On NF-κB Pathway In Spleen Deficiency Phlegm-dampness Type Non-small Cell Lung Cancer

Object iveBased on the theory of tumor microenvironment,the effects of Jianpi Huatan decotion on NF-κB signaling pathway in spleen deficiency phlegm-dampness type non-small cell lung cancer were explored from both clinical and cellular levels,Providing clinical and experimental evidence for the use of Jianpi Huatan decotion.Methods1.From April 2018 to March 2019,patients with space-occupying lesions visit cancer center in the First Affiliated Hospital of Guangzhou University of Chinese Medicine were enrolled and divided into experimental group and control group according to whether taking Chinese medicine or not.The experimental group took Jianpi Huatan decotion(JPHTF),and the control group did not take Chinese medicine.All of them received pathological biopsy and basic symptomatic supportive treatment.Eventually,patients who aren’ t pathologically diagnosed with non-small cell lung cancer will be excluded.Serum interleukin-β(IL-1 β),tumor necrosis factor-α(TNF-α),metalloproteinase-9(MMP-9),intercellular adhesion molecule-1(ICAM-1)and spleen deficiency phlegm-dampness type non-small cell lung cancer standard reference table integral(TCM score)were measured before and after treatment in both groups.The differences in TCM syndrome differentiation scores and IL-1 β、TNF-α、MMP-9、ICAM-1 between the two groups before and after treatment were compared by statistical analysis.2.With JPHTF lyophilized powder prepared,the effects of different solution on the activity of human lung adenocarcinoma A549 cells were examined at 24h,48h and 72h,and the IC50 value of JPHTF lyophilized powder dose-response curve was calculated.The basal medium was added to the control group,and the JPHTF lyophilized powder of proper concentration were added to the other group for a 72-hour incubation.The contents of IL-1 β,TNF-a,MMP-9 and ICAM-1 in cell culture supernatants were identified by ELISA.The expression of IKK β mRNA,I κ B a mRNA,NF-κ B mRNA,TNF-a mRNA,IL-1β mRNA and MMP-9mRNA,ICAM-1 mRNA levels in each group were identified by qPCR.Use scratch test to detect the migration and invasion ability of the two groups of cells.Results1.①This study included 45 patients with lung lesions at first,eventually 15 were included in the statistical analysis test group(3 excluded),and 15 in the control groupcontrol group(12 excluded).In the experimental group,8 cases were adenocarcinoma,7 cases were squamous cell carcinoma,while in the control group,14 were adenocarcinoma and 1 large cell carcinoma.As for pre-treatment age,gender,white blood cell(WBC),red blood cell(RBC),platelet(PLT),neutrophil(NEU),procalcitonin(PCT),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),the difference in two groups was not statistically significant(P>0.05).Baseline consistency was good in both groups.② Before treatment,the TNF-α,IL-1β,MMP-9,ICAM-1 and TCM score of the two groups were not significantly different(P>0.05).After treatment,serum MMP-9,ICAM-1 and TCM score in the experimental group were lower than those in the control group,and the difference was statistically significant(P<0.05).There were no significant differences in serum IL-1β and TNF-αlevels between the experimental group and the control group(P>0.05).③The serum MMP-9,ICAM-1 and TCM scores before and after treatment were much higher when compared with the control group,and the difference was statistically significant(P<0.05).There was no significant difference in serum IL-1β and TNF-a between the two groups before and after treatment(P>0.05).2.①JPHTF lyophilized powder of different concentrations could inhibit the vitality A549 cells after 24h,48h and 72h,with dependence of concentration and time.②When JPHTF lyophilized powder was applied to A549 cells for 24h,the IC50 was 20.69mg/ml;when JPHTF lyophilized powder was applied to A549 cells for 48h,the IC50 was 17.43mg/ml;when JPHTF lyophilized powder was applied to A549 cells for 72h,the IC50 was 9.753mg/ml.③The levels of IL-1β,MMP-9 and ICAM-1 in the supernatant of Jianpi Huatan decotion group were lower than those before treatment(P<0.05).There were no significant differences in TNF-a levels on the supernatant between the two groups(P>0.05).④The levels of IKK β mRNA,IκBα mRNA,NF-κ B mRNA,TNF-a mRNA,IL-1 β mRNA,MMP-9 mRNA and ICAM-1 mRNA in Jianpi Huatan decotion group were lower than those in the Control group(P<0.05).⑤The scratch test showed that invasive migration ability of A459 in Jianpi Huatan decotion group was lower than that in the control group.Conclusion1.Jianpi Huatan decotion can significantly improve the spleen deficiency and dampness syndrome of spleen deficiency and dampness type non-small cell lung cancer and reduce the levels of serum ICAM-1 and MMP-9.2.Jianpi Huatan decotion can reduce the mRNA expression of NF-κ B signaling molecules IKKβ,I κ Bα and NF-κ B and decrease the expression of downstream products IL-1β,MMP-9,ICAM-1 and their mRNA,thereby inhibiting the growth,invasion and migration of non-small cell lung cancer cells.