| The First PartHedyseri Radix belong to the petrosa Astragalus of Leguminous plant, the root of Hedysarum polybotrys Hand.-Mazz., with Astragali Radix sibling different genus, both of which have the effect of benefiting vital energy and nourish the blood and other. Hedyseri Radix substitute Astragali Radix often in traditional Chinese medicine clinically. With continuous investigation on Hedyseri Radix, we gradually discovered that Hedyseri Radix and Astragali Radix is not false indigo herb, both not only in appearance, and different in composition and content. From the1985edition of the Chinese Pharmacopoeia, people will Hedyseri Radix and Astragali Radix separate, Hedyseri Radix is single-row to a species Chinese material medical. Studies of protophase found that aqueous extract of Hedyseri Radix and Astragali Radix or hedysarum polysaccharide and astragalus polysaccharides on the immune function there are also differences. And the complex prescription is the major format of clinical applications in prevention and cure disease of traditional Chinese medicine. Comparative Study on the regulatory role in the immunity of the compound levels of Hedyseri Radix and Astragali Radix have not been reported, so we chose the classic prescription with effect of strengthening body resistance, firm appearance and tonification, Astragali Radix as principal drug, included in the2010edition of Chinese Pharmacopoeia Yupingfeng, Fuqi hidroschesis, Buzhongyiqi, and Yiqiyangxue prescription, as the object of study. The same dose of Hedyseri Radix replaced Astragali Radix, compared difference of immunomodulatory effects on immunosuppressed mice with Hedyseri Radix or Astragali Radix of each compound recipe. It is making up for the lack of pharmacological studies and medicinal theory of Hedyseri Radix, solve the deficiencies of medicinal herbs resource of Astragali Radix, proved theoretical support for comprehensive development of medicinal value and applied generalization of Hedyseri Radix.Firstly, preparation of immune suppression model was that mice injected with cyclophosphamide (CY), experimental results showed that CY mice, a variety of commonly used index to reflect the innate and adaptive immunity had declined on CY group mice, indicating that the immunosuppressive mice models were successful.Secondly, while injection of CY, with Yupingfeng, Fuqi hidroschesis, Buzhongyiqi, and Yiqiyangxue aqueous extract containing Hedyseri Radix or Astragali Radix was given simultaneously, and then index was detected:①Each group containing Hedyseri Radix or Astragali Radix could significantly improved the thymus and spleen index, the ability of peritoneal macrophages phagocytized toluylene red in vitro, content of serum IL-1β, cytotoxic activity of NK cells on immunosuppressive mice; among the effect of elevate spleen index was stronger on Yupingfeng, Buzhongyiqi, and Yiqiyangxue group containing Hedyseri Radix.②Each group containing Hedyseri Radix or Astragali Radix could improve the lever of hemolysin, percentage of CD19+B lymphocyte and content of serum IL-4, they are repressed by CY.③Each group containing Hedyseri Radix or Astragali Radix could improved to varying degrees T lymphocyte proliferative response, percentage of CD3+, CD4+and CD8+T lymphocytes subsets, content of serum IL-4and IFN-y on immunosuppressive mice; among the effect of enhance proliferation of T lymphocytes, and effect about enhance percentage of CD3+T lymphocyte and content of serum IFN-y was stronger on Yupingfeng, and Yiqiyangxue group containing Hedyseri Radix.It was concluded that:①Complex prescription to using Hedyseri Radix and Astragali Radix had a similar role in immune regulation.②Complex prescription about Yupingfeng, Buzhongyiqi, and Yiqiyangxue to using Hedyseri Radix enhance cellular immunity was better than using Astragali Radix on immunosuppressive mice slightly.The Second PartIn recent years, with the improvement of living standards and people's awareness of the disease in depth, the average human life span was significantly prolonged, while the aging of the population accompanied by serious social and medical problems, anti-aging is imminent. In the mechanism of aging, immunosenescence is the the important reasons of senescent acceleration. Among the relationship between T lymphocytes and aging have constitute the core content of the theory of immunosenescence. Aging can delay by regulating cellular immunity. The first experimental results show that it exist differences in the same compound recipe containing Hedyseri Radix and Astragali Radix, as principal drug, on immune regulatory role, and the regulatory role of T lymphocytes, of the Yupingfeng complex prescription is more obvious. It is further to select Yupingfeng for research compound recipe, to observe by in vitro experiments the effect of Yupingfeng prescription containing Hedyseri Radix and Astragali Radix on T lymphocytes of the natural aging mice, and to explore the effect and mechanism of the anti-immunosenescence.Firstly, containing drug serum with Yupingfeng to using Hedyseri Radix and Astragali Radix was prepared, than the spleen lymphocyte proliferation test was used to determine the best drug-containing serum dilute strength was40%. After corresponding index was observed in follow-up test with40%drug-containing serum and spleen lymphocytes co-cultured.①Drug-containing serum could enhanced spleen lymphocyte's seif-proliferation and ConA-induced proliferative capacity, percentage of CD3+CD28+T lymphocyte subsets and content of IL-2of the culture supernatant; and lower percentage of CD3+CD28-T lymphocyte subsets of the natural aging mice.②Through transmission electron microscopy, DNA agarose gel electrophoresis, Hoechst33258fluorescence staining and flow cytometry, spleen lymphocyte apoptosis was detected in each group, results showed that apoptosis of spleen lymphocytes of aging mice was increased, given drug-containing serum could lower the spleen lymphocyte's apoptosis. Calcium concentration of intracellular was detected by Fluo-3AM fluorescent probe's staining, results show that intracellular calcium was increased in the spleen lymphocytes of aging mouse, after given drug-containing serum, the intracellular calcium was not diversify obviously.Further expression of Fas, FasL, Bax and Bcl-2on mouse spleen cells was observed by immunocytochemistry, mitochondrial membrane potential was detected by use JC-1fluorescent probes, and Caspase3activity was detected through spectrophotometry to reflect apoptotic pathway of death receptor-mediated pathway and the mitochondrial pathway. The results showed that in the aging mouse's spleen lymphocytes, expression of Fas, FasL and Bax was increased, expression of Bcl-2was decreased; mitochondrial membrane potential was decreased and caspase3activity was increased; given with drug-containing serum could decreased the expression of Fas, FasL and Bax, could increased the expression of Bcl-2and mitochondrial membrane potential, decreased caspase3activity; among with Yupingfeng serum to using Hedy seri Radix, the effect of elevate mitochondrial membrane potential was slightly better than containing Astragali Radix group.③Used of proteomics analysis techniques, the differences of proteinum's spectrum of a graph in spleen lymphocytes of mouse about aging and given with drug-containing serum to using Hedyseri Radix or Astragali Radix group was analyzed. Expression of a variety of protein emerged changes in aged mouse spleen lymphocytes. Used PDQuest8.0software,14differences proteinum spots were selected of which analyzed by mass chromatographic, a total of nine were successfully identified. Expression of5protein was up-regulated and4protein was down-regulated in Yupingfeng drug-containing serum to using Hedyseri Radix, expression of3protein was up-regulated and6protein was down-regulated in Yupingfeng drug-containing serum to using Astragali Radix.Conclusion:①The role of spleen T lymphocytes in aging mouse was weaken②Drug-containing serum with Yupingfeng to using Hedyseri Radix and Astragali Radix both could delay aging by regulating the role of T lymphocytes.③Incidence of apoptosis was increased of spleen lymphocytes cultured in vitro in the aging mouse.④Drug-containing serum with Yupingfeng to using Hedyseri Radix and Astragali Radix both could lower the occurrence of apoptosis of spleen lymphocytes in vitro in the aging mouse. And the effect of regulating of mitochondrial membrane potential was slightly better using Hedyseri Radix group than using Astragali Radix group.⑤Successfully identified nine differentially expressed proteins may be closely related to aging, given Drug-containing serum with Yupingfeng to using Hedyseri Radix and Astragali Radix is co-cultured, and to varying degrees reversible changes in which part of the protein.
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