| Background and Objective:Lung cancer is one of the highest malignancy of morbidity and mortality in the worldwide. U.S. statistics, there are about219,440people suffering from lung cancer in2009in the United States, and nearly159,390people die of the disease. Each year dies of lung tumors about1.3million people the whole world. For the lung large frequently occurring conceal, the majority of patients are diagnosed at advanced stage and lost the chance of operation. Currently, the treatment of malignant tumors not only be treated by surgery, but chemotherapy and other means, With the development of molecular biology and pharmacology, biological therapy, targeted therapy and multidisciplinary treatment has become a hotspot of anti-tumor research.And a large number of data confirmed that the anti-cancer chemotherapy drug side effects, and long-term using is easy to produce drug resistance and other disadvantages. In recent years, Chinese medicine antitumor more and more concerned about the domestic and international pharmaceutical industry. Induced apoptosis in tumor cells as the main objectives and means of treatment of neoplastic disease. Fuzheng xiaoai mixture is a party to the mentor many years of experience.clinical studies have shown that it can improve the clinical symptoms of patients with advanced lung cancer, enhance immune function, improve quality of life. However, the specific mechanism of action is not clear.The present study, serum pharmacological method to study the Fuzheng xiaoai Mixture of apoptosis in human lung adenocarcinoma A549and explore the mechanism may exist, the possibility of theoretical and experimental basis for its clinical application.Methods:60male SD rats were randomly divided into6groups (n=10, weight is not different between the two groups):Fuzheng xiaoai mixture t low-dose group, Fuzheng xiaoai mixture dose group, Fuzheng xiaoai mixture of high-dose group,DDP group, Chinese medicine combined with DDP group, saline group, Medicines Board in accordance with the human and mouse body surface area conversion method and experimental method of serum pharmacology. They were given Fuzheng xiaoai mixture low-dose group, dose group, high dose gavage liquid medicine orally, DDP group given intraperitoneal injection0.5mg/ml DDP, traditional Chinese medicine and DDP group to righting eliminate cancer agent in dose gavage liquid medicine orally and peritoneal intragastric injection0.5mg/ml DDP, saline group to0.9%sodium chloride solution. Continuous administration of7days,2times a day, each time2ml. After2h after the last administration chloral hydrate intraperitoneal injection of anesthesia, sterile conditions, the abdominal aorta blood collection, separation of serum,56℃inactivated30min,0.22μm filter sterilization,-20℃and stored. Preparation of rat serum containing MTT assay Fuzheng xiaoai mixture containing serum on human lung adenocarcinoma A549cell proliferation. PI method flow cytometry containing serum on A549cell cycle; the Annexin V-FITC/PI double staining flow cytometry. Containing serum on apoptosis of A549cells. Western-Blot detection of serum containing P53, Bcl-2and Survivin protein expression.Results:1. Normal A549cells were adherent growth, multi-spindle or oval translucent and plump cytoplasm, cell proliferation speed observed under an inverted microscope.48h after the the righting of Xiao’aiping mixture containing serum, visible blank control group cells showed good growth. Middle dose group, part of the round cell morphology, cell gap slightly widened, the high-dose group and DDP group, the smaller cell size, cell gap is wider, and a small number of cells appear to fall off. The combined group significant cell shedding some cell swelling, fracture, were necrotic state.72h significantly reduced the number of cells, apoptosis is more significant.2. different concentrations of traditional Chinese medicine containing serum on human lung adenocarcinoma A549cells24h,48h,72h, significant inhibition of proliferation of the cells. With the increase in the serum concentration of the drug and the time extension, the inhibitory effect also will be enhanced, and time and dose-related inhibition rate and high concentration groups after DDP group, the combined group the highest inhibition rate up to38.68%, and control group, the difference was statistically significant (P＜0.05). The inhibitory rate declines, the role96h. The best time and concentration of the drug effection is can be Observed.3. Western blot test results compared with the blank control group, with the increase in drug concentration, bcl-2protein and Survivin protein gradually decreased (P＜0.05) and P53protein tended to increase (P＜0.05), and the high-dose group. Expression is higher than DDP group.Conclusion:1. Fuzheng xiaoai mixture could inhibit the proliferation of A549cells in the time and dose-dependent manner and have sy nergies combined with DDP.2. The herb serum can induce apoptosis of A549cells.early stage apoptosis accounted main part.3. Fuzheng xiaoai mixture could probably induced apoptosis of A549cells by inhibiting the expression of Bcl-2,Survivin protein,while activation of the p53protein. |
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