The Effect Of Yiqi Yangxue Decoction Containing Radix Astragali Or Radix Hedyseri On The Immune Function In SAMP8 Mice

1.The effect of Yiqi Yangxue decoction containing Radix Astragali or Radix Hedyseri on the immune function in SAMP8 mice.Objective: Three group of young mice were given equal volume Yiqi Yangxue decoction containing Radix Astragali, Yiqi Yangxue decoction containing Radix Hedyseri, and normal saline respectively to prepare medicated serum, then under the condition of co-culture of the medicated serum and SAMP8 mice spleen lymphocytes, calculating the in vitro proliferation of SAMP8 mice spleen lymphocytes, cytokine levels in the culture supernatant, differentiation of T lymphocyte subsets, the expression of p38 m RNA and p38 protein inside spleen lymphocytes, in order to compare the effects of Radix Astragali-contained Yiqi Yangxue decoction medicated serum and Radix Hedyseri-contained Yiqi Yangxue decoction medicated serum on the immune function of spleen lymphocytes in SAMP8 mice.Methods:30 Kunming species young mice of clean grade, half male and half female, were randomly divided into 3 groups with 10 in each group. Radix Astragali-contained Yiqi Yangxue decoction(6g·kg-1), Radix Hedyseri-contained Yiqi Yangxue decoction(6g·kg-1), and same volume normal saline were administered respectively to three groups by gavage once per day for unintermittent 14 days, then 2 hours after the last gastric perfusion, drawing blood through ophthalmectomy for the medicated serum preparation. The Radix Astragali-contained Yiqi Yangxue decoction group, the Radix Hedyseri-contained Yiqi Yangxue decoction group, and the blank serum group respectively co-cultured with SAMP8 mice spleen lymphocytes at the optimal medicated serum culturing concentration and time. The young mice group and the SAMP8 mice group using a complete culture medium containing 10% fetal bovine serum cultured respectively with spleen lymphocytes by their own kind. For each group, analyzing the differentiation of T lymphocyte subsets by flow cytometry, the level of IL-2 and IFN-γ in the culture supernatant with Elisa, detecting the expression of p38 m RNA inside spleen lymphocytes with real-time fluorescence quantitative PCR, and calculating the content of p38 protein inside spleen lymphocytes by Western-Blot.Results:Compared with the young mice group, the SAMP8 group significantly decreased its in vitro proliferation capacity of spleen lymphocytes, and on the differentiation of T lymphocyte subsets, the percentage of CD8+T cells decreased, the percentage of CD4+T cells increased, CD4+/CD8+ ratio increased, the percentage of CD152+T cells increased, the IL-2 and IFN-γ level of culture supernatant decreased, the expression of p38 m RNA and p38 protein in spleen lymphocytes decreased; therefore has a statistical significance of p<0.05. Compared with the blank serum group, the medicated group significantly increased its in vitro proliferation capacity of spleen lymphocytes, and on the differentiation of T lymphocyte subsets, the percentage of CD152+T cells decreased, the IL-2 and IFN-γ level of culture supernatant increased, the expression of p38 m RNA and p38 protein in spleen lymphocytes increased; therefore has a statistical significance of p<0.05. Compared with the Radix Astragali contained Yiqi Yangxue decoction group, the Radix Hedyseri contained Yiqi Yangxue decoction group is stronger in increasing its in vitro proliferation capacity of spleen lymphocytes, lesser in the percentage of CD152+T cells, a more IL-2 level of culture supernatant, and a higher expression of p38 m RNA; therefore has a statistical significance of p<0.05. Conclusion: Yiqi Yangxue decoction containing Radix Astragali or Radix Hedyseri can both promote the in vitro proliferation capacity of spleen lymphocytes, improve the differentiation of T lymphocyte subsets, increase the IL-2 and IFN-γ level, enhance the expression of p38 m RNA and p38 protein inside spleen lymphocytes. Moreover, on the aspect of boosting the in vitro proliferation capacity, IL-2 level and p38 m RNA expression, the Radix Hedyseri group is superior to the Radix Astragali group.2. The effect of Yiqi Yangxue decoction containing Radix Astragali or Radix Hedyseri on the immune function in SAMP8 mice.Objective: Respectively give SAMP8 mice and young mice the same volume of Yiqi Yangxue decoction containing Radix Astragali, Yiqi Yangxue decoction containing Radix Hedyseri, thymosin, and normal saline to get blood and spleen, then analyzing the cytokine level of serum in SAMP8 mice, the differentiation of T lymphocyte subsets, and the expression of p38 m RNA and p38 protein inside spleen lymphocytes, in order to compare the effects of Radix Astragali-contained Yiqi Yangxue decoction medicated serum and Radix Hedyseri-contained Yiqi Yangxue decoction medicated serum on immune function in SAMP8 mice and to explore the related mechanism.Methods: 10 Kunming species young mice were obtained and divided as the young mice group, 40 SAMP8 mice were randomly divided into 4 groups: the thymosin group, the saline group, the Radix Astragali group and the Radix Hedyseri group. Normal saline, thymosin(4.6mg · kg-1),Radix Astragali contained Yiqi Yangxue decoction(6g·kg-1), Radix Hedyseri contained Yiqi Yangxue decoction(6g·kg-1) were administered respectively to these groups by gavage once per day for unintermittent 14 days, then 2 hours after the last gastric perfusion, getting blood and spleen for the preparation of medicated serum and suspension of spleen lymphocytes. For each group, analyzing the IL-2 and IFN-γ level of serum with Elisa, the differentiation of T lymphocyte subsets by flow cytometry, detecting the expression of p38 m RNA inside spleen lymphocytes with real-time fluorescence quantitative PCR, and calculating the expression of p38 protein inside spleen lymphocytes by Western-Blot and IHC.Results: Compared with the young mice group, the saline group showed: the IL-2 and IFN-γ level of serum decreased, on the differentiation of T lymphocyte subsets, the percentage of CD8+T cells and CD28+T cells decreased, CD4+T cells increased, the percentage of CD152+T cells increased, the expression of p38 m RNA and p38 protein in spleen lymphocytes decreased; therefore has a statistical significance of p<0.05. Compared with the saline group, the thymosin group, the Radix Astragali group and the Radix Hedyseri group were all capable of enhancing the IL-2 and IFN-γ level of SAMP8 serum, the thymosin group is more effective than others, the Radix Hedyseri group’s IL-2 level is superior to the Radix Astragali group. The thymosin group, the Radix Astragali group and the Radix Hedyseri group were all capable of increasing the percentage of CD28+Tcells, decreasing the percentage of CD152+Tcells, the thymosin group and the Radix Astragali group have lower percentage of CD152+Tcells than the Radix Hedyseri group. The Radix Astragali group and the Radix Hedyseri group were both capable of enhancing the percentage of CD8+Tcells, and the Radix Astragali group is higher than the other. The thymosin group and the Radix Astragali group were both capable of decreasing the percentage of CD4+Tcells. The thymosin group, Radix Astragali group and Radix Hedyseri group were all capable of enhancing the p38 m RNA expression, and the thymosin group is superior to others, the Radix Astragali group is superior than the Radix Hedyseri group. The thymosin group, the Radix Astragali group and the Radix Hedyseri group were all capable of enhancing the content of p38 protein and it’s positive rate, and the thymosin group is superior to others;therefore has a statistical significance of p<0.05.Conclusion: Yiqi Yangxue decoction containing Radix Astragali or Radix Hedyseri, via enhancing the IL-2 and IFN-γ level of serum in SAMP8 mice, can both improve the differentiation of T lymphocyte subsets, the expression of p38 m RNA and p38 protein inside spleen lymphocytes, and delay the immunosenescence in SAMP8 mice. Moreover, on the aspect of increasing the percentage of CD8+T cells, both Yiqi Yangxue decoction groups are superior to the thymosin group, and on the aspect of increasing the IL-2 level of serum, the Radix Hedyseri group is better than the Radix Astragali group.