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Studies on Anti-tumor and Affecting Immune Cells Activities of Flavonoid Compounds from Chrysosplenium Nudicaule Bunge

Author: ZuoYanPing
Tutor: MaXingMing
School: Lanzhou University
Course: Immunology
Keywords: Bare stem rumped Flavonoids K562 K562/ADM SGC-7901 Apoptosis Lymphocytes Eosinophils
CLC: R29
Type: Master's thesis
Year: 2007
Downloads: 136
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Abstract


Objective: To study Tibetan part of flavonoids extracted bare stem rumped anti-tumor activity and immune cell activity, to provide experimental basis for clinical application of these compounds. Methods: (1) by MTT colorimetric assay DTFG proliferation of K562 cells and gastric cancer cell SGC-7901, TTF proliferation of leukemia the resistant cells K562/ADM and gastric cancer cell SGC-7901. (2) detection TTF vitro induced apoptosis of gastric cancer cell SGC-7901. Morphological changes were observed by fluorescence microscopy, transmission electron microscopy ultrastructural changes in the cells using flow cytometry TTF on gastric cancer cell SGC-7901 cell cycle distribution, the formation of DNA using agarose gel electrophoresis to detect apoptosis fragment. (3) the establishment of the guinea pig type I hypersensitivity model, collection of blood collected peritoneal fluid application lymphocytes stratified liquid separation lymphocytes, Percoll discontinuous density gradient separation EOS. (4) MTT the colorimetric assay DTFG and TTF guinea pig lymphocytes and EOS activity. Results: 1. Different the concentration DTFG role after the leukemia cell line K562 and gastric cancer cell line SGC-7901, within 96h did not show a significant inhibitory effect (P> 0.05). 2. Different concentrations of TTF role in leukemia resistant cell K562/ADM 72h showed different degrees of inhibition with time, TTF inhibitory effect on cell proliferation gradually increased, the inhibitory rate up to 52.82%. Drug effects different time 24h, 48h, 72h, 96h IC 50 300.31mg / L, 170.28mg / L, 65.69mg / L, 19.81mg / L. 3. Different concentrations of TTF role in gastric cancer cell SGC-7901 cell growth after 48h are subject to different degrees of inhibition. Within 72h, the concentration is greater than 4 mg / L of TTF, with time, the OD value gradually decreased, compared with the control group, the difference was statistically significant (P <0.01), while the inhibition rate gradually increased up to 80.36%. Pairwise comparison, the OD value of the difference was statistically significant (P <0.05) between the groups. Drug effects different time 24h, 48h, 72h, 96h IC 50 142.17mg / L, 29.23mg / L, 8.33mg / L, 9.75mg / L. 4. Concentration of 8 mg / L TTF role in gastric cancer cell SGC-7901 48h after observed under fluorescence microscopy and transmission electron microscopy to apoptotic cell characteristics; the TTF effect of different concentrations of gastric cancer cell line SGC-7901 after 48 h, flow cytometry cell cycle, the results show the process of cell division is blocked at the G 0 / G 1 of obvious peak of apoptosis, apoptosis rates of up to 40.4%, Compared with the control group did not receive drug treatment difference was statistically significant (P <0.01); DNA gel electrophoresis DNA ladder. 5. DTFG role in guinea pig blood lymphocytes, 72h OD value decreased compared with control group (P <0.05); role of lymphocytes in the peritoneal fluid, starting at 48h, OD value decreased compared with control group (P <0.05). The DTFG acting on guinea pig blood and peritoneal fluid EOS at different times measured OD values ??with the control group, the difference was not statistically significant (P> 0.05). 6. The different concentrations TTF role in guinea pig blood lymphocytes, the measured OD values ??compared with the control group, the difference was not statistically significant (P> 0.05). 4 mg / L TTF role of lymphocytes in the peritoneal fluid of guinea pigs, measured OD values ??decreased compared with control group, the difference was statistically significant (P <0.05). TTF and the guinea pig blood, peritoneal fluid EOS after 24h, 72h, measured OD values ??with the control group compared to the increased (P <0.01) different time PI greater than 1. Conclusion: 1. DTFG no inhibitory effect on the proliferation of K562 cells and gastric cancer cell SGC-7901. 2. DTFG guinea pig lymphocyte activity in vitro inhibited. 3. TTF has a certain inhibitory effect on the leukemia resistant cell K562/ADM of proliferation, has a strong inhibitory effect on the proliferation of gastric cancer cell SGC-7901. 4. TTF can induce apoptosis in gastric cancer cell SGC-7901. 5. The TTF can promote in vitro activity of guinea pig EOS.

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