Explore The Theoretical Of Study Of Primary Liver Cancer Pathogenesis,"Toxic Stasis Mutual Knot" And Experimental Study Of Tan Capsule Intervention Mechanism Of Arsenic Trioxide

Objective(1)Analysising etiology and pathogenesis of primary liver cancer and treat-ment aiming at its pathogenesis by finishing ancient literature,combining with modern technology.(2)To examine the effect of arsenic trioxide plus Tanshinone capsule on the proliferation and apoptosis of human bel-7404 cells in vitro,and further explore its potential signal transduction mechanism,evidencing pathogenesis and treatment of primary liver cancer.Methods(1)Literature Studies:We searched the ancient literature similar toprimary liver cancer,clearing the etiology,pathogene,treatment and drug.(2)Exprimental Studies:?human bel-7404 cells were cultured in four different drug groups,ie control group;single-drug groups with arsenic trioxide or Tanshinone capsule respectively in varied concentrations;combination group.CCK-8 analysis was used to detect proliferative effect and Annexin V-FITC/PI method was applied for apoptosis analysis in different groups.?the expressions of apoptosis related proteins including Bcl-2,Bax,Survivin,XIAP,cleaved caspase-9,cleavedcaspas-3,cleaved PARP were detected via Western blot.?Potential signal transduction mechanism of apoptosis-inducing effect in combination group was further studied by analyzing different phosphorylation levels of proteins in MAPK signaling pathway,which was also confirmed by applying different blocking agents of MAPKResults(1)The earliest medical monograph "Yellow Emperor",has a similar record of primary liver cancer.attacked by six climate pathogenic factors,improper diet and Overstrain,damaged by excess of seven emotions on the basis of stuffed to cancer,to lack of righteousness for internal conditions,phlegm and blood stasis each knot to form tumors.liver is dirty blood,liver being substantial yin and functional yang,possession and transfer of blood serum,liver tumors and blood stasis relationship is particularly close." Poisonous" "stasis" " deficiency" throughout the entire process of development of liver cancer,liver canc er pathogenesis constitute complexity.Treatment should grasp the principal contradiction,obver the pathogenesis,reating the toxifying diseases with poisonous agents,Activating blood and dissolving stasis,combined with modern science and technology research,arsenic trioxide combined with Tanshinone capsule can significantly suppress the proliferation of liver cancer.(2)?compare to the control group,The inhibition rate of bel-7404 cell after treated by 2.5,5,10,20?g/mL Tan capsules(P<0.01).the combined group(Tan capsules5?g/mL+arsenic trioxide2?m/L)on bel-7404 cell for 24h is 52.6±3.7%,significantly different compare to momotherapy groups(P<0.01).At the meanwhile this effect was in direct proportion to time,the combined group cell for 24h can not reduce suppression on LO2.On the other side the apoptosis ration were 49.4%in 24h after treated by the combined group(P<0.01).?the combination drug use can significantly down-regulate expression of inhibitors of apoptosis as Bcl-2?Survivin and XIAP,up-regulate pro-apoptotic proteins as cleaved caspase-9,cleaved caspase-3 and cleaved PARP(P<0.01).?All those effect may obtained through activating JNK,p38MAPK signaling pathway.the combined group treated bel-7404 cell after 24h,the expression of p-JNK and p-p38 were up-regulated(P<0.01),comparing to the control group and monotherapy group.and the expression of p-ERK were insignificance(P>0.05).Further study shows that JNK inhibition which treated the bel-7404 cell in 2h can inhibition the protein expression of JNK and the apoptosis protein expression of bel-7404 cell induced by Tan capsules and arsenic trioxide.Conclusion(2)Poisonous,stasis and deficiency are the most important factors on Primary Liver Cancer,which constitute complexity pathogenesis.deficiency of vital qi,cancer drug,stagnation of qi and blood stasis always throughout the liver cancer,which reduce the principle of treatment,reating the toxifying diseases with poisonous agents and Activating blood and dissolving stasis.(2)Tanshinone capsule combined with arsenic trioxide achieved synergies on human bel-7404 cells in vitro,which may be related to mitochondrial pathway,and JNK,p38MAPK pathway activation.Chemotherapy of efficient medicine combined low toxicity will induce apoptosis by multi-target and multi-pathway,which can be the theoretical basis for improving the clinical efficacy of low doses of arsenic trioxide;mechanisms of synergies revealed by modern molecular biology techniques will both clarify pathogenesis in Primary Liver Cancer,and provide methods and ideas on treating liver cancer.