The Analysis Of The Potential And Advantages Of Ethnic Medicine In The Prevention And Treatment Of Tumors And The Study Of The Regulatory Mechanism Of Tujia Cockroaches On The Apoptosis Of Liver Cancer Cells Based On The P53-GADD45? Pathway

The 21st century is a century in which life science and traditional ethnic medicine develop together.It is a new strategic goal of cancer prevention and treatment in the 21st century to search for drugs and prescriptions that can enhance human immunity,induce apoptosis of cancer cells and resist metastasis of cancer in ethnic medicine.At present,in the field of cancer disease prevention and treatment,a large number of literature studies,pharmacological experiments and clinical studies have proved to us that ethnic medicine is playing an active role.However,previous studies mostly focused on the prevention and treatment of cancer medicine of a certain ethnic group,and no article has discussed the potential and advantages of the prevention and treatment of cancer of ethnic medicine in China as a whole.Through collecting,sorting out,summarizing and summarizing the anti-cancer theory,clinical application and pharmacological research of national medicine in our country in the process of literature research,this research group aims to find common points among them and provide theoretical basis and scientific basis for the prevention and treatment of cancer of national medicine.The content of this study is divided into five main points:The foreword introduces the research background,significance and methods chosen;the first chapter,through the introduction of unique opinions of the four major ethnic medicine and Zhuang and Tujia medicine on the name of cancer and its etiology and pathogenesis,preliminarily elaborates the theoretical understanding of the prevention and treatment of cancer in ethnic medicine;the second chapter,mainly through the prevention of disease before disease-strengthening the body and resisting pathogens,prevention of disease after disease-prevention of deterioration and so on.The third chapter analyses the potential and advantages of national medicine in the treatment of tumors from the aspects of modern pharmacological research and clinical application.The summary summarizes the potential and advantages of national medicine in the prevention and treatment of cancer in China,and points out the shortcomings of this paper.Objective:To establish a rat model of cockroach extract containing drug serum and extract drug-containing serum.The effects of different drug concentration containing serum and routine western medicine containing serum on the proliferation and apoptosis of hepatocellular carcinoma BEL-7402 cells were observed by cell experiments.Using P53-GADD45? axis and its related factors ATM,CDC 2,Cyclin B1,CDK1 entry point,through the modern medical detection methods such as CCK-8,flow cytometry,Western blot,immunohistochemistry,etc.The effect on the proliferation and apoptosis of hepatocarcinoma cells and its possible mechanism of action,in order to make it an important drug for the prevention and treatment of liver cancer,but also proves that ethnic medicine has potential and advantages in the field of cancer prevention and treatment.Methods:Fifty SPF Wistar male rats were randomly divided into normal group(Normal),5-FU positive control group(5-FU),drug-containing serum high-dose group(Fei Lian High,FLH),and drug-containing serum.Medium-dose group(Fei Lian Medium,FLM),drug-containing serum low-dose group(Fei Lian Low,FLL),10 in each group.Normal group was given 10 ml/kg saline by gastric lavage;5-FU group was given 10 ml/kg 5-FU injection diluted with 0.005 g/ml pure water by intraperitoneal injection;FLH,FLM and FLL groups were given 0.5 g/ml,0.25 g/ml,0.125 g/ml cockroach extract solution 10 ml/kg by gastric lavage respectively.The rat model of cockroach extract containing drug serum was established twice a day for 5 consecutive days.One hour after the last administration,3%pentobarbital was anesthetized by intraperitoneal injection,abdominal aorta blood was taken 7-8 ml/rat.After 1 hour,the complement was inactivated by centrifugation for 10 minutes at 3500 r/min,supernatant was taken at 56 degrees Celsius water bath for 30 minutes,and the drug-containing serum was filtered and packed separately by 0.22 micron filter.The serum was stored at-20 degrees Celsius and thawed when needed.The proportion was prepared by cell culture liquid.The BEL-7402 cells were cultured with the serum containing drugs at the required concentration.The corpses and wastes of laboratory animals should be treated harmlessly according to ethical requirements.Pathological observation:(1)CCK-8 was used to detect the proliferation of hepatocellular carcinoma cells;(2)Flow cytometry was used to detect the cell cycle distribution of hepatocellular carcinoma cells;(3)Flow cytometry was used to detect the apoptosis of hepatocellular carcinoma cells;(4)Immunohistochemistry was used to detect the expression of P53,GADD45? and Cyclin B1 in hepatocellular carcinoma cells;(5)Western blot was used to detect the expression of ATM,P53,GADD45?,Cyclin B1 and CDK1 in hepatocellular carcinoma cells.Results:(1)The results of CCK-8 assay showed that compared with Normal group,5-FU group had the most obvious inhibitory effect on the proliferation of hepatocellular carcinoma cells(P<0.01),followed by FLH group(P<0.01).The inhibitory effect of FLM on proliferation of hepatocellular carcinoma cells was significant at 72 hours(P<0.01).(2)Flow cytometry showed that compared with Normal group,the proportion of G2/M,G1/G0 phase cells increased in 5-FU group and FLH group(P<0.01),while the proportion of S phase cells decreased(P<0.01).(3)Flow cytometry showed that compared with Normal group,5-FU group had the most obvious effect on promoting apoptosis of hepatocellular carcinoma(P<0.01).FLH,FLM and FLL groups all showed the effect on promoting apoptosis of hepatocellular carcinoma(P<0.01),especially in high dose group.(4)Immunohistochemistry showed that mutant P53 and Cyclin B1 were strongly positive in Normal group(+++),5-FU group showed the most obvious inhibition(P<0.05),FLH group took the second place(P<0.05),GADD45? was low in Normal group,high in 5-FU group(P<0.01),followed by FLH group(P<0.05).(5)Western blot results showed that the expression levels of ATM,P53,CyclinB1 and CDK1 in hepatocellular carcinoma cells in 5-FU,FLH and FLM groups were significantly lower than those in Normal group(P<0.01),while the expression levels of P53,CyclinB1 and CDK1 in FLL group were significantly lower(P<0.05);compared with Normal group,the expression levels of GA in hepatocellular carcinoma cells in 5-FU,FLH and FLM groups were significantly lower(P<0.05).Compared with Normal group,the expression level of GADD45? protein in hepatocellular carcinoma cells in 5-FU,FLH and FLM groups was significantly higher(P<0.01),and the expression level of FLL histone protein was higher than that in Normal group(P<0.05).Conclusion:(1)Tujia cockroach extract can promote the apoptosis of hepatocellular carcinoma cells,which may be achieved by inhibiting the proliferation of hepatocellular carcinoma cells and mediating cell cycle arrest.(2)Tujia cockroach extract containing serum can down-regulate the expression levels of ATM,P53,Cyclin B1 and CDK1 in BEL-7402 cells and up-regulate the expression level of GADD45a protein,suggesting that cockroach extract may promote apoptosis of hepatocellular carcinoma cells by regulating P53-GADD45? pathway and related signal factors.