The Expression And Pathway Of Rab30 In PC12 Cells Under The H₂O₂ Intervention

Objective This paper investigates the expression of Rab30 in PC12 cells under the intervention of H2O2, The signal pathways of Rab30 in PC12 cells are also discussed preliminarily.Methods 1、The PC12 nerve cells are purchased from Chinese academy of sciences/Cell bank of Shanghai, which are nourished in a constant temperature(37oC) carbon dioxide incubator with saturated humidity of 5% and PH of 7.2~7.4. The PH is regulated by using complete medium(containing 10% fetal bovine serum, 90% 1640 culture, and 100 U/ml green-streptomycin solution). When the cells with convergence rate of 80%, they are digested and passaged by using trypsin, and the cells in logarithmic growth phase are selected as the research objects. 2、Two experimental groups are set up: normal group and H2O2 group. 3、The Annexin V-FITC/PI double staining is used to examine the apoptosis rates of each group. 4、The Western-blot technique is used to examine theprotein expression of Rab30, JNK3, GM130, Caspase-3.Results 1、Under the microscope, PC12 nerve cells present the characters of multiple angle shape, unstable adherence, forming cell condensation easily, halation surrounded, strong diopter, growing in clumps, forming a dense cluster and covering the whole incubation bottle after 5 to 7 days nourishment. 2、Using MTT method to determine the survival rates of PC12 cells, we find the absorbance value of H2O2 group itreated is obviously lower than that of normal group(P<0.05). Compared with the normal group(0.2553±0.0084), the absorbance value A of H2O2 group(0.1577±0.0100) is lower(P<0.05), with statistical significance. 3、Using Annexin V-FITC/PI double staining to examine each group, we find the apoptosis rates of H2O2 group is significantly increased. The early apoptosis rates and the total rates of the H2O2 group(32.7500±1.0485,46.1167±1.1501) are significantly increased compared with normal group(0.5333±0.0216, 0.6333±0.0450), with statistical significance(P<0.05). 4、Using Western-blot technique to examine the expression of protein, we find JNK3 and Caspase-3 in the injury treated groups are significantly increased, while Rab30 and GM130 are decreased, compared with the normal group. The expression of Rab30 in H2O2 group(0.0570±0.0014) is embodied statistical significance(P<0.05),compared with the normal group(0.2272±0.0020). The expression of JNK3, GM130 and Caspase-3 in H2O2 group(0.6904±0.0211, 0.0821±0.0038 and 0.8620±0.0198, respectively), are also embodied statistical significance(P<0.05).Conclusion 1、H2O2 can induced the injury or apoptosis of the PC12 cells. 2 、 The expression of Rab30 in the PC12 cells is downgrade under the H2O2 injury intervention. 3、H2O2 can downgrade the expression of Rab30 and GM130(through JNK3), break the golgi into pieces, and induce the apoptosis of the PC12 cells through adjusting the expression of Caspase-3.