Effect Of Substitution Of Radix Hedysari For Radix Astragali In Yupingfeng San On Anti-immunosenescence Of T Spleen Lymphocytes Of SAMP8 Mice

objective:We adopted the fast aging (SAMP8) animal model, and determined T lymphocytes proliferation in spleen of mice, T lymphocytic percentages CD3+, CD4+, and CD8+ in spleen of mice, the levels of IL-2 in cultured supernatant of lymphocytes in spleen of mice, as well as the expression levels of CD28 mRNA and p38 MAPK mRNA in the signal transduction pathway of CD28, to compare with the effects of the same compound using Radix Astragali (RA) and using Radix Hedysari (RH) on immunologic function of mice. And then to explore the mechanism and the similarity of RA and RH in Yupingfeng San in anti-immunosenescence function.Method:1. The 60 normal Kunming mice were randomized into three groups with 20 in each, i.e. the blank group, Yupingfeng San containing RA group, Yupingfeng San containing RH group. Yupingfeng San containing RA and RH groups were given Yupingfeng San containing RA and RH 4 g/kg/d respectively, while the blank serum group was given 0.9% saline. After the last administration and 1 hour later, removed the eyeball to prepare the contaning medicated serum, which was divided into different concentrations and cocultured with spleen lymphocytes of SAMP8 mice. We detected the best concentration and optimum culture time of medicated serum with the methyl thiazolyl tetrazolium (MTT) assay.2. We cocultured medicated serum and the spleen lymphocyte of SAMP8 mice, and detected its own proliferation ability and increment ability with ConA inducing by MTT method after the red stilbazolium Yupingfeng San containing serum and Astragalus Yupingfeng San containing serum effected on spleen lymphocytes of SAMP8 mice in vitro culture, and then, we detected CD3+, CD4+ and CD8+ in T lymphocytes by flow cytometry method, detected content changes of IL-2 in cell supernatant by enzyme linked immunosorbent assay (ELISA) method.3. We cocultured the best dilution concentrations of serum and spleen lymphocytes of SAMP8 mice, and then detected expression levels of CD28 mRNA and p38 MAPK mRNA in spleen T lymphocytes of SAMP8 mice by real-time fluorescent quantitative (RT-PCR) method.Result:1. MTT results showed that the spleen lymphocytes proliferation activity increased gradually (P<0.01) whether using RA or RH to be compared with the same concentration blank serum group,40% of drug concentration with serum training 48h stimulated proliferation best acting on spleen lymphocyte in SAMP8 mice when Yupingfeng San containing Radix Astragali and containing Radix Hedysari acting on spleen lymphocyte of SAMP8 mice.2. Compared with the blank serum group, YuPingfeng San whether using RA or RH were able to make spleen T lymphocyte proliferation of SAMP8 mice promoted (P<0.01), Stimulation index was increased (P<0.01); lymphocytic percentages of CD3+CD4+and CD3+CD8+T lymphocyte subsets promoted (P<0.01), the ratio of CD3+CD4+/CD3+CD8+significantly increased (P<0.05), content of IL-2 in cell supernatant increased (P<0.01).3. Compared with the blank serum group, expression levels of CD28 mRNA and p38 MAPK mRNA of the spleen T lymphocytes increased (P<0.01). Among them, the spleen lymphocyte proliferation with RA in Yupingfeng San promoted significantly to compare with RH (P<0.05), content of IL-2 in cell supernatant with RA in Yuping Feng San increased significantly (P<0.05).Conclusions:Yupingfeng San containing Radix Hedysari or Radix Astragalis both have the effect of anti-immunosenescence through regulating the T lymphocytes, but Yupingfeng San containing Radix Hedysari has better effect on T lymphocytes proliferation and IL-2 level than Yupingfeng San containing Radix Astragali.