| ObjectiveThe purpose of this study is to establish the rat model of aging caused by the D-galactose and to study the mechanism of Qiongyugao, the salve used to maintain life. There is detected the 8-OHdG, the iconic product of oxidative DNA damage, and the related genes or protein of the MAPK/ERK1/2 signal pathway and discusses the mechanism of Qiongyugao acting on the spleen of D-galactose-induced aging rats by regulating the MAPK/ERK1/2 signal pathway.MethodsForty-two 6-month-old male SD rats of SPF level were randomly divided into three groups (Control group, Model group, and QYG-experiment group). The rats of model group and QYG-experiment group are intraperitoneal injection D-galactose (125mg/kg) to manufacture a senile rat model every day. The rats of control group are treated with the same dose of normal saline. Simultaneously, the rats of QYG-experiment group are garaged the Qiongyugao oral solution (20ml/kg)and the rats of control group and the model group treated with the same dose of distilled water. All the rats are sacrificed and dissected the spleen to prepare the samples after 62days.It is to detect the expression of SIRT3,8-OHdG, NF-K B and P-ERK1/2 by IHC, and to detect the expression of BCAR1, P-ERK1/2 by Western blotting, and to detect the expression of BCAR1mRNA, P-ERK1/2mRNA by PCR.Results1. D-galactose induced sub acute aging rats spleen tissue IHC results:(1) The expression of SIRT3 in all the groups of rats is positive. The expression of SIRT3 in the rats spleen tissue of control group and the model group is lower than the QYG-experiment group.(2) The expression of 8-OHdG in all the groups of rats is positive. The expression of 8-OHdG of model group is the highest, while the QYG-experiment group is lowest.(3) The expression of NF-K B in all the groups of rats is positive. The expression of NF-K B of QYG-experiment group is significantly lower than the model group and the control group.(4) The expression of P-ERK1/2 in all the groups of rats is positive. The expression of P-ERK1/2 of QYG-experiment group is significantly lower than the model group and the control group.2. D-galactose induced sub acute aging rats spleen tissue Western blotting results:(1) Compared with control group, the expression of BCAR1 of the model group is significantly increased (P<0.01). And compared with the model group, the QYG-experiment is inhibited obviously (P<0.05)(2) Compared with control group, the expression of P-ERK1/2 of the model group is significantly increased (P<0.05). And compared with the model group, the QYG-experiment is inhibited obviously (P<0.05)3. D-galactose induced sub acute aging rats spleen tissue PCR results:(1) Compared with control group, the expression of BCAR1 mRNA of the model group is significantly increased (P<0.05).And compared with the model group, the QYG-experiment is inhibited obviously (P<0.05)(2) Compared with control group, the expression of P-ERK1 mRNA of the model group is significantly increased (P<0.01).And compared with the model group, the QYG-experiment is inhibited obviously (P<0.05)(3) Compared with control group, the expression of P-ERK2 mRNA of the model group is significantly increased (P<0.05).And compared with the model group, the QYG-experiment is inhibited obviously (P<0.01). Conclusion1. Qiongyugao can pretect the spleen tissues of D-galactose-induced aging rats by Oxidation protection. In addition, it can effectively reduce the level of oxidative DNA damage in rat spleen cells.2. Qiongyugao can effectively increase the level of expression of SIRT3 and reduce the level of expression of P-ERK1/2,NF-κB,BCAR1 and the transcriptional level of BCAR1 mRNA and P-ERK1/2 mRNA.3. This experimental study shows that the Qiongyugao can inhibit the MAPK/ERK1/2 signal pathways play, reduce the activity of NF-K B and then inhibit the inflammatory response and cell apoptosisa.4. Qiongyugao plays a role of MAPK/ERK1/2 signaling pathway may have two kinds of paths, including SIRT3-ERK1/2 pathway and BCAR1-ERK1/2 pathway. |
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