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The Study of Antitumor Activity of Oxytropis Kansuensis Bunge Alkaloid Fraction and Its Immunomodulation Activity on Mouse

Author: WangMingJuan
Tutor: LiMin；ZuoDuoLong
School: Lanzhou University
Course: Pathology and Pathophysiology
Keywords: Gansu Oxytropis Swainsonine Human hepatoma HepG2 cells Suppression / tumor rate Cell cycle arrest Apoptosis H22 hepatoma cells Immune function
CLC: R730.5
Type: Master's thesis
Year: 2009
Downloads: 69
Quote: 0
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Abstract


Purposes: (1) in vitro research Gansu alkaloid parts inhibitory effect on human hepatoma HepG2 cells; observed alkaloid fraction of HepG2 cells in cell cycle arrest and induction of apoptosis; detection alkaloid fraction of HepG2 cells morphology vitro migration, invasion and adhesion ability. (2) The in vivo experimental observation Gansu Oxytropis alkaloid fraction of the the KM mouse hepatoma H 22 inhibition of tumor cell transplantation and KM mice immune function; observed Gansu alkaloid parts extremely high-dose oral toxicity. Methods: (1) In vitro inhibition rate of Gansu Oxytropis alkaloid fraction of human hepatoma HepG2 cells and HepG2 cells and collagen type IV adhesion ability was detected by MTT; flow cytometry alkaloid fraction of HepG2 cells induced role in apoptosis and cell cycle arrest; inverted phase contrast microscopy alkaloid fraction of HepG2 cell growth and morphology; of Boyden chambers detection alkaloid fraction of HepG2 cell migration, invasion. (2) in vivo experiments using murine hepatoma cells H 22 the transplanted tumor Act, 60 KM mice were randomly divided into five groups, three groups were given Gansu Oxytropis the alkaloids parts 32,16 , 8 mg / kg body weight, the two control groups were given normal saline (NS) of 20ml/kg weight and 5-FU 15mg/kg body weight. Detection average tumor weight and tumor inhibition rate calculated spleen index (SI), thymus index (TI), MTT method to measure spleen cell proliferation rate. High dose oral toxicity test given 3000mg/kg the alkaloid fraction biggest volume weight, the mice were observed toxicity. Results: (1) Gansu Oxytropis alkaloid fraction can inhibit the growth of human hepatoma HepG2 cells in a time-and dose-dependent, statistically significant compared with the control group (P <0.05). (2) flow cytometry Gansu Oxytropis alkaloid fraction induced HepG2 cell cycle arrest in a dose-dependent manner; small doses can induce apoptosis in HepG2 cells. (3) Boyden chamber in vitro migration, invasion capacity test results showed Gansu Oxytropis alkaloid fraction of HepG2 cell migration, invasion, inhibit the ability was statistically significant (P <0.01), compared with the control group. (4) the adhesion experiment results show alkaloid fraction of Gansu Oxytropis inhibit the adhesion of HepG2 cells with collagen type IV, was statistically significant (P <0.05) compared with the control group. (5) In vivo experiments Gansu Oxytropis alkaloid fraction at concentrations as 32,16,8 mg / kg body weight, 22 hepatoma cells H entity that tumor inhibitory rate: 43%, 29 %, 23%, and the average tumor weight compared with the control group was statistically significant (P <0.01). (6) spleen index and thymus index of Gansu Oxytropis alkaloid fraction of each dose group were significantly higher than that of the control group, there was statistically significant (P <0.01). (7) in each group of Gansu Oxytropis alkaloid fraction 24h, 48h, 72h spleen cell proliferation rate was significantly higher than that in the control group, a statistically significant (P <0.05). (8) high-dose oral toxicity test results show that, compared with the control group, Gansu Oxytropis alkaloid fraction of the weight and quality of major organs of KM mice had no significant effect was not statistically significant (P> 0.05). Conclusion: (1) Gansu Oxytropis alkaloid fraction in vitro significantly inhibit the growth of human hepatoma HepG2 cells in a time and dose-dependent manner. (2) the Gansu oxytropis alkaloid fraction can induce liver cancer cell cycle arrest in HepG2 cells in a dose-dependent manner; and in small doses can induce apoptosis in HepG2 cells, suggesting a possible anti-tumor mechanism. (3) Gansu Oxytropis alkaloid fraction inhibited HepG2 cells in vitro migration, invasion and adhesion to collagen type IV, suggesting that the alkaloid fraction by changing the biological behavior of HepG2 cells to suppress the metastatic ability of tumor cells. (4) Gansu Oxytropis alkaloid fraction the the KM mouse hepatoma H 22 transplanted tumor inhibition. (5) Gansu Oxytropis alkaloid fraction significantly enhanced the KM mouse spleen index, thymus index and spleen cell proliferation rate, suggesting that they have enhanced the role of the immune function of mice. (6) in Gansu Oxytropis alkaloid fraction high dose oral toxicity test prompted alkaloid fraction oral administration without significant toxicity.

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