Transfer Gas Consumer Product Tang Experiments On Mice With Lewis Lung Tumor Effect

Purpose:Lung cancer is one of the common malignant tumors in China, it threat human health and life seriously. For the lung cancer characteristics, based on years of clinical practice, we have to use Tiaoqixiaoji Decoction in the clinical treatment of lung cancer to achieve a good effect.For"air-lift"theory of Traditional Chinese medicine as a guide, for Lewis lung cancer of mice as a study, we discuss the possible mechanism of Tiaoqixiaoji Decoction in mice bearing Lewis lung cancer cells, at general life status,the inhibition ratio of tumor,immunologic function,apoptosis and tumor angiogenesis in this experiment, in order to provide a scientific theoretical basis for the treatment of lung cancer. Material and method:Inoculated the forty-eight C57BL/6 mice with Lewis lung cancer cells, after 5 days choosing successful tumor-bearing mice, then randomly divided into four groups: Model group(MG),Tiaoqixiaoji Decoction group(BG),DDP (Cisplatin) group(DG)and Tiaoqixiaoji Decoction-DDP combination group,10 mice in each group; other ten healthy mice were control group(CG).Different drug were given to five groups: Drinking water,Tiaoqixiaoji Decoction at 0.4ml once a day by gastrogavage, total 12 days;Normal sodium,DDP at 0.13ml every other day by intraperitoneal injection, total 3 days . Then to observe the daily life and the tumor growing status of mice; H.E.staining was used to observe the pathological changes of tumor tissue by optical microscope. To observe the effect of the Tiaoqixiaoji Decoction on the index of spleen and count the T lymphocyte transformation rate of the peripheral blood by morphology method; The T lymphocyte proliferation of spleen of all group mice were determined by MTT colorimetric determination; To detect the expression of tumor necrosis factor-α(TNF-α)by Western Blotting method. Determination of cell apoptosis of tumorous tissue were performed by electron microscope (EM)and Tunel method; The expression of mutant-type p53,Bcl-2 of tumor tissues were determined by Western Blotting method. To detect the expression of Microvascular density(MVD),Matrix metalloproteinases-9(MMP-9) and Vascular endothelial growth factor(VEGF) by Western Blotting method.Results:1. From the body weight,food intake,mice reactivity point of view, the Tiaoqixiaoji Decoction could improve the general status of the Lewis tumor-bearing mice. The times of autonomic activities and standing within five minute were increased, there were statistical difference compared with the MG(P<0.01), and no significant difference between the TG & CG(P>0.05).2. Compared with the MG, there were some cell morphological changes of the TG,DG and ZG. Such as the volume of some tumor cells were smaller and the abnormal cells became less, the cell nuclei were tightening, the Chromatin were condensed in margin, and the pathological karyokinesis reduced obviously; Lymphocytes infiltrated in interstitial and seldom tumor cells can be seen in capillaries, surrounding necrosis area increased.3. The weight of tumors of the TG,DG and ZG were much lower than the MG,and there were significant difference compared with the MG(P<0.01), the inhibition ratio were 29.37%,71.23% and 75.19%. It shows that the Tiaoqixiaoji Decoction could inhibit growth of tumors and the inhibitory effect appears obviously when Tiaoqixiaoji Decoction combines with DDP.4. The spleen index of the MG were significantly increased to 111.54% of the CG's, there were statistical difference between the TG & CG(P<0.01); and the spleen index of the TG were closed to normal value, there was no statistical difference compared with the CG(P>0.05); the DG and ZG spleen index decreased significantly, they were 36.55% and 54.62% of the CG respectively.5. From the T lymphocyte transformation rate of the peripheral blood and the T lymphocyte proliferation of spleen of all group mice point of view, the MG was obviously decreased to the CG, and there were significant difference(P<0.01 or P<0.05); but the TG was higher than the MG(P<0.01),and there was no statistical difference compared with the CG(P>0.05); the DG was the lowest,and there were significant difference compared with the CG,MG. Say that the Tiaoqixiaoji Decoction can improve the immunologic function of tumor-bearing mice.6. The high expression of TNF-αwas showed in Lewis lung tumor tissue of the DG and ZG by Western Blotting method,there were significant difference compared with the MG(P<0.01); but there were no statistical difference between the TG and MG(P>0.05).7. Through the observation of EM, we found the specific signs of apoptosis-- apoptotic bodies.8. In TUNEL situ assay showed, the apoptotic cells of the MG were rare and the green fluorescence intensity is low; increased apoptotic cells could be seen in all treatment groups, the green fluorescence intensity is high; the ZG was highest. It shows that the Tiaoqixiaoji Decoction and DDP combined could enhance the ability to promote apoptosis in lung cancer cells.9. Western Blotting results showed, the TG,DG and ZG all could reduce the expression of mutant-type p53 of tumor tissue, the ratio of the grey levels toβ-actin were : (0.752±0.006),(0.723±0.008),(0.721±0.011),there were statistical difference compared with the MG(0.777±0.008) (P<0.05 or P<0.01); the expression of mutant-type p53 of the ZG was lowest.10.Western Blotting results showed, compared with the MG, the expression of MVD,VEGF and MMP-9 decreased obviously in all treatment groups, and there were obviously difference(P<0.05 or P<0.01); and there were no statistical difference between the TG and DG in the expression of MVD,VEGF and MMP-9(P>0.05).Conclusion:1. The Tiaoqixiaoji Decoction is effective in improving the general life status of tumor-bearing mice,and it can inhibit growth of the Lewis lung cancer. If combine with DDP, it can enhance the ability of anti-tumor.2. The Tiaoqixiaoji Decoction can protect the immune organ of Lewis lung cancer in mice; it can improve the immunologic function of tumor-bearing mice by increasing the T lymphocyte transformation rate of the peripheral blood and the T lymphocyte proliferation of spleen; and can not effect expression of TNF-αin tumor tissue.3. The Tiaoqixiaoji Decoction can induce the apoptosis of the Lewis lung cancer cells, if combine with DDP, it can enhance the ability to promote apoptosis; its activity of inhibiting the growth of tumor may be relative to decreasing the expression of mutant-type p53 and Bcl-2.4. The Tiaoqixiaoji Decoction can degrade the MVD of Lewis lung cancer; and also can reduce the expression of VEGF and MMP-9 in order to play the effect of inhibiting tumor angiogenesis, anti-tumor.