[Effects of Hesperetin on cell activation and proliferation of murine T lymphocytes in vitro].
Yuying Zhou, Yaoying Zeng, Xiu−Yan Huang, Bing Song
Abstract
Yuying Zhou, Yaoying Zeng, Xiu−Yan Huang, Bing Song
Abstract
AIM: The effect of Hesperetin (Hes) on activation and proliferation of murine T lymphocytes in vitro as well as its mechanism of action is investigated to provide a theory for developing an immunosuppressive agent. METHODS: The lymphocytes were cocultured with Concanavalin A (ConA) and Hes together. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 of activated T lymphcytes in vitro in response to ConA. MTT test was used to estimate the effect of Hes on proliferation of lymphocytes stimulated by ConA and the toxic effect of Hes on lymphocytes. Proliferation of lymphocytes stimulated by ConA was detected by carboxyl fluorescein diacetate-succinimide ester (CFDA-SE) staining combined with flow cytometry, and the proliferation index (PI) was analyzed by means of ModFit software. RESULTS: The survival rate of lymphocytes shows that DMSO(0.2 mL/L)and Hes (25-75 micromol/L) had little side effect on murine lymphocytes in vitro; Hes can inhibit the activation and proliferation of T lymphocytes in response to ConA with noticed dosage-effect relation. CONCLUSION: Hes may has broad prospects to be developed as immunosuppressive drug through further studies.
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AIM: The effect of Hesperetin (Hes) on activation and proliferation of murine T lymphocytes in vitro as well as its mechanism of action is investigated to provide a theory for developing an immunosuppressive agent. METHODS: The lymphocytes were cocultured with Concanavalin A (ConA) and Hes together. Fluorescence conjugated monoclonal antibodies and flow cytometry were used to detect the expression of CD69 of activated T lymphcytes in vitro in response to ConA. MTT test was used to estimate the effect of Hes on proliferation of lymphocytes stimulated by ConA and the toxic effect of Hes on lymphocytes. Proliferation of lymphocytes stimulated by ConA was detected by carboxyl fluorescein diacetate-succinimide ester (CFDA-SE) staining combined with flow cytometry, and the proliferation index (PI) was analyzed by means of ModFit software. RESULTS: The survival rate of lymphocytes shows that DMSO(0.2 mL/L)and Hes (25-75 micromol/L) had little side effect on murine lymphocytes in vitro; Hes can inhibit the activation and proliferation of T lymphocytes in response to ConA with noticed dosage-effect relation. CONCLUSION: Hes may has broad prospects to be developed as immunosuppressive drug through further studies.
Key concepts: Concanavalin A, Flow cytometry, In vitro, Molecular biology, Hesperetin, Mixed lymphocyte reaction, Chemistry, Cell growth