The Effect Of Jiedufang On The Inflammatory Response After Hepatic Artery Chemoembolization Of Primary Liver Cancer And Its Molecular Mechanism

Objective:The present study was a prospective cohort study conducted in order to explore the clinical curative effect of Jiedu Fang(JDF)on Hepatocellular carcinoma(HCC)patients treated by Transcatheter arterial chemoembolization(TACE).We also analized the serum inflammatory factors change before and after TACE and tried to find out the possible mechanism.Methods:1.Enrolled HCC patients treated with TACE in Changhai hospital from February 2018 to August 2018.Divided them randomly into JDF Group(treated with TACE and JDF)or Control Group(treated with TACE).Assessed with MD Anderson symptoms scale before and after TACE to explore the influence of JDF and TACE on the quality life of the HCC patients and evaluated whether JDF may alleviate postoperative embolism syndrome.Peripheral blood was drawn before,3 days and 2 months after TACE to detect the levels of inflammatory factors such as blood routine,liver function,Interleukin-1β,and to analyze the changes of inflammatory factors in the two groups and the changes based on the inflammatory scoring system.Finally,we tried to find out the possible inflammatory factor that was the target of JDF.2.Kaplan-meier survival curve was drawn and log-rank test was performed to compare the difference of Progresstion free survival(PFS)between the JDF group and the control group.Meanwhile,COX proportional risk model was used to analyze the possible prognostic factors of PFS at 6 months after TACE,and independent influencing factors were screened out.3.MTT assay was used to detect the effect of JDF on the proliferation of EA.hy 926 cells,and angiogenesis experiment was used to detect the effect of JDF on angiogenesis under hypoxia.PCR experiments were applied to explore the effects of JDF on the Hypoxia inducing factor-1α(HIF-1α),Vascular endothelial growth factor(VEGF)and IL-8 m RNA expression level.ELISA was used to further explore the IL-8 expression under hypoxia and treated with JDF.Western blot assay was used to detected the effects of IL-8 on HIF-1α and VEGF.Finally,angiogenesis assay was used to detected the effects of IL-8 and JDF on the angiogenesis.4.A matrix gel angiogenesis model was established to verify in vivo the effects of il-8 on matrix gel tissue hemoglobin concentration,VEGF,CD31 and v WF,as well as the antagonistic effects of JDF.Results:1.A total of 88 patients with HCC were included in this study,with 47 in the control group and 41 in the JDF group.GPS and m GPS scores of all the enrolled patients were higher after than before TACE.NLR increased from preoperative(2.56±2.48)to postoperative(8.50±4.40),and PLR increased from preoperative(110.66±49.46)to postoperative(136.26±78.04).levels of IL-1β,IL-2R,IL-6 and IL-8 were all up-regulated.Compared with the control group,JDF can effectively improve the quality of life of HCC patients after TACE,in which pain,vomiting,constipation are the most obvious improvement.The complete response rate of the JDF group for TACE postoperative embolism syndrome was better than that of the control group,which may be related to the reduction of IL-8,PLR and NLR.Among which the inhibition of IL-8 was the most obvious.2.In the 2-month short-term efficacy evaluation,the objective response rate(ORR)of the JDF group was 41%,the disease control rate(DCR)was 71%,and the ORR of the control group was 13%,the DCR was 68%.The JDF group was superior to the control group.JDF group showed superior on the progression-free survival(PFS)than that in the control group at a 6-month follow-up.Univariate COX survival analysis showed that PLR before TACE,IL-2R on day 3 after TACE,and m GPS,GPS,PLR on 2 months after TACE were the influencing factors.Multivariate analysis found that m GPS,PLR and IL8 before TACE,NLR,IL-2R,AFP on day 3 after TACE,and PLR on 2 months after TACE were all factors affecting PFS.3.MTT experiment results showed that EA.Hy 926 cell proliferation was inhibited in a time-dependent and concentration-dependent manner after treated with JDF at different concentrations for 24 h or 48 h,but the inhibition rate was <20%.RT-PCR results showed JDF could inhibit the overexpression of IL-8,HIF-1α and VEGF m RNA in Huh 7 cells induced by hypoxic condition.ELISA assay also demonstrated the effect of JDF on IL-8 expression.Both hypoxia and IL-8 may promote angiogenesis and JDF may inhibit their effects,suggesting that JDF may inhibit angiogenesis by inhibiting the expression of IL-8 under hypoxia conditions,thus exerting anti-liver cancer effect.4.In vivo matrix gel angiogenesis assay showed that IL-8 could promote matrix gel tissue angiogenesis and upregulate the expression of hemoglobin.HE staining results showed that JDF could antagonize IL-8-induced angiogenesis.Immunohistochemical results showed that the positive expression rates of VEGF,CD31 and v WF in the IL-8 group were significantly higher than those in the control group,while the positive expression rates of VEGF,CD31 and v WF in the il-8 +JDF group were all lower than those in the IL-8 group,suggesting the antagonistic effect of JDF on IL-8.Conclusion:1.JDF combined with TACE can significantly reduce the expression of inflammatory factors in HCC patients after TACE and improve the quality of life of HCC patients and prolong their PFS.2.JDF inhibits the proliferation of Huh 7 hepatocellular carcinoma cells and inhibits angiogenesis under hypoxia in vitro,which may be related to the inhibition of IL-8,HIF-1 and VEGF expression.In vivo,JDF can effectively inhibit the IL-8-induced angiogenesis.