| Objective:To cultivate C6 glioma rat source cells in vitro.Establish SD rats C6 gliomas in situ transplanting animal model.To study the growth regulation effect and potentiai mechanism of rat C6 glioma cell in body by pseudomonas aeruginous MSHA (PA—MSHA). To explore new method of effectively postoperative adjuvant therary for glioma. Methods:To cultivate C6 glioma rat source cells in vitro. Choose male health SD rats 30, the weight of body between 200g±20 g..To establish the SD rat C6 glioma allograft animal model by stereotactic method. We conducted a non-blinded, randomized, controlled trial.40 rats with glioma were randomly assiganed to receive nothing or the left by PA—MSHA vaccine (experimental group). Determine the cell vitality, collect logarithm in good condition of growth C6 glioma cells, adjusting cell concentration to 1.0×107/ml,inject one rat with 1.0×107 cells. Positioning right-brain subcortical areas with animal brain stereotactic meter(beside the midline 2mm, before opening Xin 3mm), inject 20 u 1 to each animal.After 7 days, inject rats with 0.9% Nacl and PA—MSHA 0.1ml. View the general state of the tumor-bearing mice after administration; Observe the value-added rate of tumor and living time. After that we used HE and TEM to study the glioma and Flow Cytometry to investigate the Immune function before and after treatmeat at 7th day control group and 20th day experimental group.Results:1.To establish the SD rat C6 glioma allograft animal model C6 glioma cells used in this experiment,. the survival rate were always over 95%.Behavior significantly change in rats after 7 days of transplantation. Only 20 model are established successfully.2.View the general state of the tumor-bearing rats The control group rats have less food and water than the experimental group after vaccination. Contract the experimental group, the control group showed decreased activity and body weakness, increase the incidence of symptoms such as limb paralysis by the time..3.Used HE and TEM to study the glioma. All the rats were resected the tumor under anesthesia in the 20 days after inoculation, The growth C6 glioma cells of the experimental group in lower density than the control group and some regions appear in liquefaction necrosis. by HE, and can see the smaller size of cell contraction, nuclear pyknosis deep-dyed blue morphological changes of apoptosis cells by TEM, showing early apoptotic changes.4.Flow cytometry to investigate the Immune function. Collected the serum at different time by the flow cytometry, The level of the CD3,CD4 and CD8 in the control group are lower than normal group (P<0.05) The level of the CD3,CD4 and CD8 in the experiment group are higher than control group (P<0.05)Conclusion:PA-MSHA preparations can suppress the proliferation of C6 glioma cells. The mechanism may be related apoptosis and enhance immune functions, especially the cellular immunity are incressed significantly. It can also activate cytokine network in the cell line.
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