The Academic Experience Of Professor DouYong-Qi On Treating Tumor And Study On The Mechanism Of HuanQi-Ezhu Formula On Anti-Tumor Angiogenesis

Objective:To summarize the clinical experience and academic thought of Professor DouYong-qi on treating tumor by the method of combination of diseases differentiation and syndromes differentiation.Through the animal and cell experiments to study the anti-tumor effects and anti-angiogenic mechanism of compatibility of HuanQi(HQ)and Ezhu(EZ)formula which were the common TCM herbal drugs used in combination by tutor.Methods:1.Through analyzing and summarizing the tutor's theory explanation,clinical teaching,academic dissertations and knowledge lectures,clinical experience and the rules of administering medicines were analyzed.Combining with the discussions and the review of TCM literature,the tutor's academic thought of combining diagnosis and treatment of malignant tumors were summarized.2.LLC bearing tumor mice models were randomly divided into model group,cisplatin(DDP)group,HQ group,EZ group,and 1:1,2:1,3:1 groups of compatibility of HQ and EZ.The TCM groups were given the corresponding doses of drugs for once a day;and the cisplatin group was given cisplatin(2mg/kg)for once two days.All mice were sacrificed on the 16th day;the tumor weight index,tumor inhibition rate,the number of lung surface metastatic nodes,thymus and spleen index were calculated;and the pathological changes of the tumor tissues were analyzed by H-E staining.The microvessel density(MVD)of tumor tissues were analyzed by the immunohistochemistry(IHC)staining.Western blot was employed to confirm the expression of key factors of TGF-?1/MAPKs/HIF-la signaling pathway.3.A uniform design was used to optimize the optimal combination(E)from the 4 active ingredients of HQ and EZ herbs.MTS assay was applied to measure the A549 cell proliferation inhibition rate(CPIR).The inhibitory effect of E on the proliferation of A549 cells under different concentrations and time conditions was observed.A549 cells were exposed to CoCl2(200 ?mol/L)to mimic the hypoxic conditions.Blank group was treated with RPMI-1640,the group CoCl2 was treated with CoCl2,the group DDP+CoCl2 was treated with 4mg/L Cisplatin injection(DDP)+CoCl2,and the drug group was treated with various dose of E(0.5E,1E 2E)+CoCl2.Cell apoptosis rate was measured by flow cyometry.Western blotting was employed to confirm the protein expression of caspase-3,Bax,Bcl-2,and the expression of key factors of TGF-?1/MAPKs/HIF-la signaling pathway.Results:1.Tumors have a common basis of pathogenesis and similar pathological mechanisms,and moderm medical treatment can lead to different pathological changes in the body.The different development stages,different clinical features and different treatment methods of malignant tumor are included in the category of "identifying disease".Tutor treat the malignant tumors with the method of "combination of disease identification and syndrome differentiation".The tutor emphasized that the TCM should participate in the whole process of malignant tumor with the combination of western medicine in a dynamic and individualized form.2.Tumor weight of the administration groups were lower than model group(P<0.05).The tumor weight index and the lung metastasis nodules number in 3:1 group were no significant difference(P>0.05)to compare with the DDP group.The thymus and spleen indexes of all TCM groups were higher than DDP group(P<0.01).Compared with the model group,the MVD expression of the administration groups were significantly decreased(P<0.01),the MVD expression of 3:1 group was the closest to the DDP group.The expression of TGF-?1 in 2:1 and 3:1 groups were significantly decreased(P<0.01).Compared with the model group,the expression of HIF-la on 1:1,2:1 and 3:1 compatibility group were significantly decreased(P<0.01).Compared with the model group,the expression of p38MAPK,p-p38MAPK,ERKl/2,p-ERK1/2,JNK and p-JNK of compatibility groups were significantly decreased(P<0.01).The VEGF protein expressions in 2:1 and 3:1 groups were significantly decreased to compare with the model group(P<0.01).3.The optimal combination of components(E)obtained by uniform design was 200mg/L Astragalus polysaccharide and 32 mg/L Curcumin.E could significantly inhibit the proliferation of A549 cells in a dose-and time-dependent manner.Compared with DDP group,the apoptosis rates of E and 2E groups were not significantly different(P>0.05).Compared with DDP group,the expression of Bax and Bcl-2 in 2E group was not significantly different(P>0.05),while the caspase-3 expression in each E group was lower than DDP group(P>0.05).The expression of TGF-?1,HIF1? and VEGF in 2E+CoCl2 group was lower than DDPgroup(P<0.01).The expression of p38MAPK and p-p38MAPK in E and 2E groups were lower than DDP group(P<0.01),while the expression of JNK and p-JNK in 0.5E and 2E was lower than DDP group(P<0.05).The expression of ERK and p-ERK in each dose of E group was lower than DDP group(P<0.01).Conclusions:1.Tutor believe that the common pathological basis of tumor are the vital qi deficiency while pathogenic qi prevailing,qi and blood disorders;the common pathological mechanism are qi stagnation,blood stasis,sputum coagulation and carcinomatous toxin.Based on the disease differentiation and four diagnostic methods in TCM,according to the different stages of tumor and different western medical treatment methods,tumor patients are given personalized,dynamic TCM treatment,which can improve the quality of life of tumor patients and improve the level of the tumor treatment.2.The compatibility of different proportions of HQ and EZ could inhibit the tumor growth and metastasis in LLC xenograft mice mode,its mechanism might be relevant to reduce the expression of the MVD of tumor tissue,the expression of key factors of TGF-?1/MAPKs/HIF-1? signaling pathway and inhibiting the tumor angiogenesis.3.Astragalus polysaccharides combined with curcumin is the best combination of four known active ingredients in the combination of HQ and EZ,which can inhibit the proliferation and promote apoptosis of lung cancer A549 cells;which can down-regulate the expression key factors of TGF-?1/MAPKs/HIF-1? signaling pathway and inhibiting the tumor angiogenesis.