Effects Of Hawthorn Flavonoids On Immune Activity And CGMP And PKC Signaling Pathway Of Spleen Lymphocytes In Mice

Hawthorn is a food of Rosaceae,and Hawthorn flavonoids are one of the functional representative components of Hawthorn.Studies have found that hawthorn flavonoids have the functions of enhancing immunity,regulating blood lipid,dilating blood vessels,helping digestion and resisting cancer,etc.At the same time,it has the pharmacological effects of resisting oxidation,protecting brain,liver and kidney,etc.In order to explore the immunomodulatory effect of hawthorn flavonoids,this paper established a screening model of immunological activity with primary mouse spleen lymphocytes as the target cells,and discussed the immunomodulatory effect of hawthorn flavonoids and its effects on c GMP and PKC signaling pathway in vitro from the cellular and molecular levels.The experimental results are as follows:1.MTT assay was used to detect the effect of hawthorn flavonoids on the proliferation of mouse spleen lymphocytes.The results showed that hawthorn flavonoids could promote the proliferation of spleen lymphocytes in mice in the dose range of 100-800μg/m L,thus playing an immunomodulatory role.2.The effects of hawthorn flavone on the secretion of cytokines IFN-γ,IL-4,IL-6 and IL-12 by mouse spleen lymphocytes were detected by ELISA.Real-time PCR was used to detect the effects of hawthorn flavone on the m RNA expression levels of IFN-γ,IL-4,IL-6and IL-12 in spleen lymphocytes of mice.The results showed that hawthorn flavone could promote the secretion and m RNA expression of IFN-γ,IL-6 and IL-12,and inhibit the secretion and m RNA expression of IL-4 in spleen lymphocytes of mice in the dose range of100-800μg/m L,so as to play an immuno-regulatory role.3.The effects of hawthorn flavonoids on cell membrane surface markers and cell cycle of mouse spleen lymphocytes were detected by flow cytometry.The results showed that hawthorn flavonoids could increase the percentage of CD4~+,CD8~+and CD4~+/CD8~+ratio of mouse spleen lymphocytes in the dose range of 100-800μg/m L.Hawthorn flavonoids can reduce the percentage of G1 phase in mouse spleen lymphocyte cycle,and increase the percentage of entering S phase and G2 phase,so as to play an immunomodulatory role.4.The effects of hawthorn flavonoids on c GMP content and PKC activity in mouse spleen lymphocytes were detected by ELISA.The results showed that hawthorn flavonoids could activate c GMP and PKC signal pathway in mouse spleen lymphocytes in the dose range of 100-800μg/m L,thus playing an immunomodulatory role.