Gs Mixture On The Experimental Study Of The Influence Of The Lung Cancer Cells In Mouse Cachexia

Lung cancer around the world, whether male or female, is the incidence of one of the high mortality of malignant tumors, causing countries to medical treatment, significant economic losses. The current medicine although with drugs to treat lung cancer, but the negative effect is quite large, therefore, to discover and identify effective treatment for lung cancer drug with reduced toxicity, is the urgent work. Through the experimental study by the Chinese herbal medicine on lung cancer cells of tumor-bearing mice, and the observation of Chinese herbal medicine for cachexia in tumor-bearing mice provide other possible ways and direction of cancer treatment strategies and the development of new drugs.According to the study of cancer patients,87%will have symptoms of cachexia,22%of patients die from cachexia rather than the tumor itself. Therefore, to understand why, and provide effective measures to prevent and treat cancer cachexia in the cancer treatment is more important. Cachexia in advanced cancer is the common clinical disease in patients with integrated symptom, especially received radiotherapy and chemotherapy. The clinical manifestations include digestive disorders, body weight loss, fatigue, anemia and body weakness, and even a direct threat to the lives of patients. Therefore, how to improve the status of the patients with cachexia, has become a pressing problem in the clinical work.Traditional Chinese medicine to increase and stabilize weight, improve appetite and activity index, has considerable effect. Through the clearing and detoxifying, nourishing Qi, improving spleen and stomach activity, traditional Chinese medicine does can improve the quality of life of cancer patients with cachexia state. Although the use of traditional Chinese medicine (TCM) in China has a long history, and their extracts since the ancient times has a variety of treatment of disease or even a key ingredient for anti-cancer and anti-cachexia, there is still a lack of valid scientific evidence to confirm its efficacy. Therefore, how to use this active ingredient combined with establishment of closely related animal model, and to show its effectiveness is a focal point we must strive for a breakthrough.Objective:Traditional animal experiments for drug screening, most of the cancer cells are implanted under the skin, and then observed the change of the biochemical indicators, or cytokines of the mice. Hoverer, this animal model can not be close to the actual situation of general patients, we develope an orthotopic mouse animal model to carry the animal experiments and hopefully it becomes a useful tool for screening anti-cancer herbal medicines in the future.Chinese herbs with functions of both the heat clearing and detoxification are the first choice of anti-inflammatory and anti-cancer drug since ancient times. They can alleviate inflammation and cachexia status caused by the lung cancer cells. Therefore, in the previous experimental basis, we select the mixture of Lonicera japonica Thurb, Prunella vulgar is and Taraxacum mongolicum H, to evaluate the improvement of mouse cachexia caused by the lung cancer cells. And hopefully the Chinese herbal medicine with treatment of lung cancer can be effectively developed.Animal studies of lung cancer treatment in the past, cancer cells were implanted in the skin, then the amount of tumor size were measured with vernier, but orthotopic carcinoma growth situation was unable to collect the information of its growth curve. However, it has recently developed a set of in vivo imaging system using luciferase to decompose the D-lucif in in order to observe and analyze the cancer cell growth in vivo situation without sacrificing the mice, so we can continuously collect large amounts of data for anti-cancer drug screening and drug reactions of cancer cells in mice, thus can get more information. We intend to apply this technology to see if this technology can be applied to the orthotopic lung cancer mouse model.Cytokine play an important role in the human body. Most of the cell communication between cell and immune cell are carried by cytokines. Cytokine is a small molecule of protein, it can change itself or the behavior of other cells; Therefore, the amount of cytokine in the body can represent the status of the cells, or through the regulation of cytokine's level, it can affect the cell behavior, thereby regulating the immune cells of animals and affect the immune status of the whole organism. We hope that the cellular immunity as a research model can help us understand the correlation between the herbal extracts and cellular immune responses in order to clarify any relation between cytokines and Chinese herbs with promotion of immunity and anti-tumor activity, and to find out an indicator in the cell experimental stage, thus, filter out the Chinese herbal ingredients with efficacy.Methods:1. GS mixture on the experimental study of the influence of the lung cancer cells in mice cachexia.In this experiment, the orthotopic lung cancer H460cells were implanted in the chest cavity of mice to induce cachexia; the following day, the mice were fed with Chinese herbal medicine in order to observe the situation of cachexia improvement. Analysis of indicators for the mouse body weight, food intake and serum albumin, glucose, triglyceride, and survival rate of mice were the basis for this assessment. The experiment was divided into a total of six groups Normal, Model, GS, GSRS, GSRA and SG. The Chinese herbs were proportionally mixed and extracted, and each mouse was fed daily with TCM for0.2g/kg.2. Establishment of A549-Luc tumor xenograft animal model:correlation of tumor weight with bioluminescence.In this experiment, the orthotopic lung cancer A549-Luc cells were implanted in the chest cavity of mice to induce cachexia. After the implantation of the cancer cells in the mice pleural space, the luminescence intensity in the mice was measured in order to detect the growth of the tumor cells. The mice were put in the act of euthanasia and anatomy to remove the tumor and weighed. The relative curve of the luminescence intensity v.s. tumor weight was able to identify the two corresponds'relationship.3. Stimulation of cytokine production from immune cells and inhibition of U937cell growth by adding Chinese Herbal Medicine Astragalus and Codonopsis extracts.In this experiment, the cellular immunity is as a research model in order to understand the Chinese herbal medicine, Astragalus and Codonopsis extracts (DSUF3K, DSUFPS, SGHF3, HGUFPS, HGF3and HGW) and the correlation between the cellular immune response in order to clarify the relation between cytokine and the Chinese herbal medicine with raising immune and anti-tumor activity; and to further explore the effect of the condition medium, which was stimulated from PBMC by Chinese herb extracts, on U937cell line growth inhibitionResults:1. GS mixture on the experimental study of the influence of the lung cancer cells in mice cachexia.The experimental results show that the orthotopic implantation of lung cancer H460cells did induce cachexia in mice, body weight of mice and their biochemical indicators are significantly decreased. The drugs tested, GS can elavate the serum albumin concentration (GS:173.86±15.15ng/ml, Model:155.77±17.02ng/ml, p<0.05), however, there is no clear statistical significance between the experimental group and model group (P>0.05) of all other biochemical indicators.2.Establishment of A549-Luc tumor xenograft animal model:correlation of tumor weight with bioluminescence.The experimental results show that the reporter gene has been successfully implanted in the A549lung cancer cells. Established A549-Luc cells were implanted in pleural space of the mice, then, determined the luminescence performance and weight change in mice; the mice were sacrificed, and the tumor was removed and weighed. Experimental results show that tumor weight is proportional to the luminescence intensity and tumor weight greater than30mg, is a linear relationship with the luminescence intensity (when luminescence intensity is3.06±1.081,4.22±1.70,4.87±1.61,5.53±4.29x105photons/sec/cell, the tumor weight is0.03±0,0.03±0,0.17±0.02.0.28±0.05g), therefore, a similar experiment can be on this basis to measure the tumor size with the use of luminescence intensity projections the tumor size, while reducing the sacrifice of the mice.3. Stimulation of cytokine production from immune cells and inhibition of U937cell growth by adding Chinese Herbal Medicine Astragalus and Codonopsis extracts.The experimental results show that the test substance DSUF3K, DSUFPS, SGHF3, HGUFPS, HGF3and HGW all can stimulate PBMC to produce high concentrations of cytokine, and inversely proportional with the concentration.From the inhibition of the U937cell growth inhibition test, inhibited by the condition medium stimulated by the Chinese herbal medicine extract on PBMC cells, we know that the experiment shows condition medium stimulated by lower concentration of DSUF3K, DSUFPS SGHF3, HGUFPS HGF3and HGW has higher inhibition rate of U937Cell line, and the higher the concentration of the extract, the inhibitory effect is less obvious.Conclusion:1. Orthotopic carcinoma implantation of lung cancer H460cells in mice, indeed lead to mouse cachexia. Food intake, body weight, TG, glucose, and serum albumin of mice are decreased, showing the feasibility of this mouse model of cachexia. Among the test herbs, the GS has a function to enhance serum albumin (p<0.05), however, its mechanism may be required to further explore; the remaining indicators had no significant statistical meaning (p>0.05).2. In vivo fluorescence imaging of the growth of lung cancer A549cells in SCID mice can be continuously observed. We found that fluorescence can be observed after11days of implantation of the tumor cells. Experimental results show that tumor weight is proportional to the luminescence intensity and tumor weight greater than30mg, is a linear relationship with the luminescence intensity.3. Test substance DSUF3K DSUFPS, SGHF3HGUFPS, HGF3and HGW can all stimulate PBMC to produce high concentrations of cytokine and inversely proportional with the concentration, which we found that high concentrations of the extract of Chinese herbal medicine also inhibit the growth of PBMC.From the inhibition of the U937cell growth inhibition test, inhibited by the condition medium stimulated by the Chinese herbal medicine extract on PBMC cells, we know that the experiment shows condition medium stimulated by lower concentration of DSUF3K, DSUFPS SGHF3, HGUFPS HGF3and HGW has higher inhibition rate of U937Cell line, and the higher the concentration of the extract, the inhibitory effect is less obvious. This result seems directly proportional to the amount of cytokine measured; but the actual cause of this inhibitory effect remains to be further explored by which cytokine.