| Objective: To probe into the molecular mechanism of Fuzheng Yiliu Keli(FYK) in inducing cellular apoptosis of tumour by integral and isolated experiments. Methods: (1) Integral experiment: The mechanisms of the whole and separated prescriptions for inducing cellular apoptosis were observed. The mice inoculated with neoplastic cell H22 are randomly divided into 8 groups-Fuzheng group, depressing tumor group, the big dosage of FYK group, the middle dosage group, the small dosage group, Tianxian capsule group, CTX group, model group and blank control group. The mice are fed with the drugs of same volume. The apoptosis of neoplastic cell was inspected with TUNEL, the protein expression of Bax, Bcl-2 and Caspass-3 in neoplastic tissues with immuno-histochemical method and the morphological change of neoplastic cell with electron microscope. (2) Isolated experiment: The whole prescription of Fuzheng Yiliu Keli( FYK) was divided into 5 groups , the blank group, the big ,middle and small dosage group of FYK and Tianxian group. The serum contained with Chinese herbal medicine was prepared, by which the H22 cells were cultured outside the body. The changes of the numbers of living cells are observed by the trypan-blue method and the growing curve had been drawn. The restrained rates for the growth of cultured cells outside the body were detected by the MTT method. The periods of cellular propagation were examined by the technique of flow-cytometry. The protein expressions of Bcl-2, Bax, p53, Fas, FasL and Caspase-3 associated with apoptosis had been detected by the immuno-histochemical method. The changes of [Ca2+] inside the cells were examined with the laser focused microscope. 3 Results Results of integral experiments: (1) The effects of the whole prescription of FYK on depressing tumor were remarkably higher than that of the separated ones.( P<0.05) (2) The FYK can predominantly induce apoptosis of neoplastic cells and strengthen phagocytic function of macrophagus. (3) The FYK can activate the gene expression of Bax, Caspases, increase the related protein expression of Bax, Caspase-3 for accelerating apoptosis and at the same time lower the protein expression of Bcl-2 for depressing apoptosis. Results of the isolated experiment: (1) The serums with different dosages of FYK had remarkable effects on depressing the vitality of the propagation of cell H22, which is markedly different, compared with the matched control groups. (P<0.05) The depressing rates of the big, middle and small dosage group of FYK were respectively 79.3%, 54.6% and 31.3%. It shows that the larger the dosage is, the higher the depressing rate is .The rate is elevated with the dosage of FYK. (2) Results of the technique of flow-cytometry showed that the medicinal serum with FYK increased the time in the period of G0/ G1 and reduces the time in period S and period G2/ M , the cell propagation index ( PI)decreased, particularly in the middle and big dosage groups. ( P<0.05) (3) Results of the TUNEL method showed that the serum with FYK can markedly increase the apoptosis rate of cell H22. Results of the technique of flow-cytometry showed that 72 hours later after being affected by the serum with FYK, the period G1/G0 of each group using the medicine presented with the peak of apoptosis. The larger the dosage and the longer the time of being affected, the higher the apoptosis rate is. In different dosage group, cells in period G0/ G1 increased, cells in period S and G2/ M decreased, especially in middle and big dosage groups. (P<0.01) (4) In each group of different dosages, the numbers of positive Bcl-2 cells reduced and the strength of reaction decreased obviously, while the numbers of positive Bax cells increased and the strength enhanced, which was coincided with the results of the integral experiments completely. Fas expression was lower in cell H22. The serum with FYK can clearly increase Fas expression of cell H22, especially in the middle and large dosage groups, which had remarkable difference, compared with the blank control grou...
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