The Effect Clinical Efficacy Evaluation And Mechanism Of Yi Qi Huo Xue Decoction Combined With TACE Of Advanced HCC

Object:To study a method combined by Yi Qi Huo Xue decoction and TACE,observe its efficacy and explore its mechanism based on the tumor microenvironment,in order to treat HCC using a new and effective way with the integrated traditional Chinese and Western medicine.Methods:Clinical research:Among the patients in June 2018 to December 2019 with advanced HCC admitted to The Second Affiliated Hospital of Nanjing University of Chinese Medicine(Jiangsu Second Chinese Medicine Hospital).The type of TCM syndrome is the type of qi deficiency and blood stasis.By way of assessment conforms to the discharge standard,and voluntary to join the clinical studies.50 cases with Yi Qi Xu Xue Yu decoction(25 cases in the treatment group,25 cases in the control group)are suitable for intervention therapy.The control group:Conventional interventional therapy was used alone.The proper hepatic artery was searched under the guidance of DSA,and the 4F catheter was followed.According to the results of DSA,the blood supplying artery of the liver tumor was located,and then the catheter was pushed into it for related operations.For specific performance:5-Fu 1g,Oxaliplatin 100mg.After the infusion of drugs,iodinated oil was used for hepatic artery embolization to block the supplying blood vessels.4 weeks was a course of treatment,and the method was repeated after 4 weeks for 2 cycles.The treatment group:The interventional treatment was the same as the control group.On the 2nd day after intervention,the Yi Qi Huo Xue decoction was taken orally.After 8 weeks of treatment(2 cycles of treatment),a comprehensive review of all indicators was conducted.The efficacy was comprehensively evaluated by comparing the changes in the target lesions,Traditional Chinese Medicine(TCM)symptoms,biomarkers in serums of tumors(CEA?AFP?CA199),FPS,1-year survival rate,and patients' physical status,and an 8-week systematical evaluation of related toxic side effects and post-embolization syndrome(within 1 week after intervention).Experimental research:through animal experiments and in vitro experiments,to explore the mechanism of Yi qi Huo Xue decoction's intervention in HCC.? Preparaed drug-containing animal serum.Through cell tests,simulated the hypoxic internal environment after an interventional operation,observed the effect of Yi Qi Huo Xue decoction on the activity and proliferation of liver cancer cells under hypoxia,compared the invasion and migration ability of the two groups of cells using the Transwell chamber,and explored its intervention effect on the HGF/c-Met pathway in combination with the internal tumor environ ment?Through animal experiments,observed the anti-tumor effect of Yi Qi Huo Xue decoction combined with interventional therapy on liver cancer model in mice.Established the Liver metastasis model of nude mice.selected 30 nude mice and divided into 6 mice per group:Model group,simple chemotherapy drug(5-Fu+oxaliplatin)group,Yiqi Huoxue decoction high-dose+chemotherapy group(high TCM group),Yiqi Huoxue decoction medium-dose+chemotherapy group(middle TCM group),Yiqi Huoxue decoction low-dose+chemotherapy group(low TCM group).After establishing the nude mice,the first medication was given 72 hours later,qd × 2w.The high TCM group was 3.38g/mL,the medium TCM group 1.69g/mL,and the low TCM group 0.84g/mL,each 0.2mL.The model group and the chemotherapy group were given the same NS as the traditional Chinese medicine group.Chemotherapy drugs 5-Fu(diluted with normal saline)and oxaliplatin(diluted with 5%glucose solution)were diluted to concentrations of 30mg/ml and 5mg/ml,respectively,and intraperitoneal injection of 0.2ml was given to the traditional Chinese medicine group and the chemotherapy alone group.Sacrificed the nude mice 15 days later to observe liver metastases.Resected the liver metastases and some adjacent tissues.Using RT-PCR to further observe the impact of Yiqi Huoxue decoction on the expression of apoptosis-related genes Caspase-3,Survivin,Bax,and Bcl-2,and using Western blot to observe the interventional effects of anti-angiogenesis-related genes VEGF,HIF-1?,and VEGFR-2,and further explored its regulation effects of HGF/c-Met pathway in the liver cancer microenvironment.? Observe the effect of Yi Qi Huo Xue decoction on multi-drug resistance to HepG2/ADM through cell tests.Using MTT assay to detect the inhibitory effect of Yiqi Huoxue decoction on HepG2/ADM cell growth and the effect on drug sensitivity after the addition of ADM,OXA,5-Fu and Sora Feenib.Using Western blot to study its impact on P-pg proteins in the microenvironment to explore its anti-drug resistance mechanism.Results:Clinical research:45 cases were finally completed,including 22 cases in the treatment group(fall off 3 cases)and 23 cases in the control group(fall off 2 cases).Before the drug and interventional treatment intervention,the basic information of patients in the two groups was statistically analyzed,including gender,age,KPS and Child-pugh grade,which showed no significant statistical significance(P>0.05)and was comparable.1.Evaluation of solid tumors and serum biomarkers:1.Evaluation of solid tumor and serum markers:At 8 weeks after treatment,CT scans were performed to measure the size of the target lesions and compare them with baseline.In the treatment group,the effective rate was 90.9%,no CR,PR,13 cases,SD7 cases;In the control group,the effective rate was 82.6%,no CR cases,5 PRs and 17 SDs.The effective rate of the treatment group was better than that of the control group,but there was no significant difference(P>0.05).In terms of serum tumor markers,CEA,AFP and CA199 were not significantly different between the treatment group and the control group before treatment(P>0.05).After treatment,CEA,AFP and CA199 were decreased in the treatment group,but there was no statistical significance after statistical test(P>0.05).After treatment,CEA and AFP decreased in the control group,while CA199 increased slightly,with no statistical significance(P>0.05).After treatment,the levels of CEA and CA199 in the treatment group were significantly lower than those in the control group,the differences were statistically significant(P<0.05),AFP was lower than those in the control group,but there was no statistical significance(P>0.05).2.TCM symptom evaluation:The treatment group and the control group were evaluated and analyzed respectively before and 8 weeks after treatment.In the treatment group,6 cases were significantly effective,13 cases were effective,and 3 cases were ineffective,with an effective rate of 86.4%.In the control group,4 cases were significantly effective,9 cases were effective,10 cases were ineffective,and the effective rate was 56.5%.The effective rate of treatment group was higher than that of control group,with statistical significance(P<0.05).3.Evaluation of 1-year overall survival rate and time to disease-free progression:The PFS and 1-year survival rates of the treatment group were slightly higher than those of the control group,and there was no statistical difference(p>0.05).The 1-year survival of the patients was 63.6%.Compared with the control group,47.8%,there was no significant difference(p>0.05).4.Physical status and toxic and side effects:8 weeks after treatment,the KPS status of the two groups was reassessed.In the treatment group,8 cases were improved,12 cases were stable,and 2 cases were deteriorated.The stability rate(improvement+stability)was 90.9%.In the control group,7 cases were improved,7 cases were stable,and 9 cases were deteriorated,the stability rate was 60.9%;The results showed that after treatment,the stability rate of the treatment group was significantly higher than that of the control group(p<0.05).In terms of interventional syndrome,Within 1 week of interventional treatment,the incidence of fever,pain,nausea and vomiting in the treatment group were all lower than those in the control group,with statistical significance(P<0.05).There was no significant difference in the myelosuppression above grade III(including grade III)between the two groups(p>0.05);there were no significant differences in liver and kidney toxicity indexes AST,ALT,BUN,and CREA(p>0.05).Experimental Research:1.In vitro experiment:The A value in SMMC-7721 cells,cell proliferation rate,and the number of cells migrating to the lower chamber in the experimental group were lower than those in the control group(all p<0.05).The expression levels of HGF and HGF mRNA in the cell culture supernatant of the experimental group,and the expression levels of c-Met protein and mRNA in the cells were lower than those of the control group(all p<0.05).2.Animal experiment:The body weight of mice in the middle and high-dose groups of Yiqi Huoxue Decoction increased(p<0.05),which was statistically significant.Compared with the model group,the tumor weight of the high-dose group and the chemotherapy group was significantly reduced,and the difference was statistically significant.Significance(p<0.05);In terms of inducing apoptosis,Yi Qi Huo Xue decoction can increase the mRNA levels of pro-apoptotic genes Caspase-3(chemotherapy group and Chinese medicine high group)and Bax(chemotherapy group and Chinese medicine Chinese group,Chinese medicine high group)The expression is statistically significant(p<0.05),which can reduce the expression of apoptosis-suppressing genes Survivin and Bcl-2 mRNA(chemotherapy group and Chinese medicine high group),which is statistically significant(p<0.05);in terms of anti-angiogenesis,Yiqi Huoxue Decoction can reduce the expression of VEGF,HIF-1? and VEGFR-2 protein.Compared with the model group,the difference is statistically significant(p<0.05).Among them,the high-dose Yi Qi Huo Xue decoction combined with chemotherapy is better than the chemotherapy group alone.In comparison,the difference is statistically significant(p<0.05);in terms of signal pathways,the middle and high-dose combination chemotherapy group of Yi Qi Huo Xue decoction can significantly down-regulate the protein expression levels of HGF,c-Met,pc-Met,and MEK,compared with the model group There is the statistical significance(p<0.05).3.Intervene the drug resistance of liver cancer:Yi Qi Huo Xue decoction treatment of HepG2 and HepG2/ADM has increased tumor inhibition rates,which are positively correlated with time and concentration,and there is no significant difference between the two(p>0.05).The effect of HepG2/ADM is added to ADM.After OXA,5-Fu,and Sorafenib,each group's cell proliferation inhibition effect was still noticeable.The medium and high dose groups were significantly different from the control group(p<0.05),and there was no significant difference between the drug groups(p>0.05).With the increase of Yiqi Huoxue compound's drug concentration,the P-GP protein decreased,and the middle and high-dose groups showed a significant decreasing trend(p<0.05).Conclusion:Yi Qi Huo Xue decoction combined with interventional therapy can improve patients'clinical symptoms and physical status with advanced liver cancer and improve the quality of life,And can reduce the incidence of interventional syndrome.Compared with simple interventional therapy,it has certain synergistic and detoxifying effects.However,there were no obvious advantages in the changes of target lesions,tumor indicators,FPS and 1-year survival rate in a short time.In addition,the follow-up time was too short to study the benefit of OS.Compared with simple western medicine treatment,the subjective feelings of patients have been greatly improved,and the effect is significant,which reflects that this prescription has a better clinical effect and has its advantages.Cell and animal experiments again verify that Yi Qi Huo Xue decoction can resist tumor development and metastasis.It also can regulates the HGF/c-Met signaling pathway.Through the animal experiments,it can increase the body weight and decrease the tumor weight of nude mice bearing tumor.It also can reduce Survivin and Bcl-2,increase caspase-3 and Bax mRNA,and induce tumor cell apoptosis.It can down-regulates VEGF,HIF-la and VEGFR-2,and can anti-angiogenic.It can down-regulate the expression of HGF gene mRNA,c-Met gene mRNA and MEK gene mRNA in a concentration-dependent manner,which again indicates that Yiqi Huoxue Presence can interfere with the HGF/c-Met pathway.In terms of drug resistance studies,it can reduce P-gp protein,thereby inhibiting the proliferation of HepG2/ADM,and inhibit the drug resistance of ADM,OXA,5-Fu,and sorafenib.Around the tumor microenvironment,its mechanism revolves including inhibition of induced cell apoptosis,anti-angiogenesis,interventing the HGF/c-Met signaling pathway and reversing multi-drug resistance.