The Preliminary Study Of The Mechanisms Of Anti-inflammatory Of Simvastatin In Chronic Heart Failure

Objective To observe the level of expression of CD25,CD69 and HLA-DR on human T lymphocyte surface; to observe the interleukin-2(IL-2), interleukin-6(IL-6) and tumor necrosis factor(TNF-α) expression and production of human T lymphocyte; to explore the effects of simvastatin on expression of CD25,CD69 and HLA-DR of human T lymphocyte in vitro; to explore the effect of simvastatin on IL-2, IL-6 and TNF-αexpression and production of human T lymphocyte; to investigate the influence of simvastatin on NF-κB of human T lymphocyte.Methods (1) Jurkat cells were incubated with PHA with or without different concentration simvastatin in vitro. After incubation, the toxicity was detected by Tapalan test, in order to definite the safety margin of simvastatin and PHA in vitro; (2) Jurkat cells were incubated with PHA with or without simvastatin at the different concentrations in vitro, simvastatin was added two hours before PHA. After incubation 4, 24, 48, 72 hours, the cells were collected and the expression of cell markers such as CD25, CD69 and HLA-DR on cell surface was assayed with flow cytometry, in order to investigate the effect of simvastatin on CD25, CD69 and HLA-DR expression of T lymphocyte surface in vitro; (3) Jurkat cells were incubated with step 2, and the supernatants were collected and IL-2, IL-6 and TNF-αconcentrations in the supernatants of Jurkat cells were measured by radioimmunoassay(RIA); (4) Jurkat cells were incubated and collected with step 2, and the levels of IL-2, IL-6 and TNF-αmRNA were quantified with real time fluorescent quantitative polymerase chain reaction(RQ-PCR), in order to investigate the effect of simvastatin on the transcription of IL-2, IL-6 and TNF-αmRNA; (5) Jurkat cells were incubated and collected with step 2, and the transportion of NF-κB into the nuclear was detected by immunocytochemistry.Results (1) There was no significant toxicities on examined Jurkat cells when simvastatin was used at concentrations less than or equal to 10μM, or when PHA was used at concentrations of 5μg/ml; (2) Simvastatin could significantly inhibit the expression of CD25 and CD69 on T lymphocyte surfaces (p<0.05), and the higher the simvastatin concentration, the more serious inhibition of the expression of CD25 and CD69 on T lymphocyte surfaces. The inhibition was the most serious after 24 hours or 48 hours; (3) The expression and production of IL-2, IL-6 and TNF-αwas inhibited by simvastatin (p<0.05), and the higher the simvastatin concentration, the more serious inhibition of IL-2, IL-6 and TNF-αexpression and production. The inhibition was the most serious after 24 hours or 48 hours; (4) Simvastatin could significantly inhibit the transportion of NF-κB into the nuclear (p<0.05), and the higher the simvastatin concentration, the more serious inhibition of the transportion of NF-κB into the nuclear. The inhibition was the most serious after 24 hours or 48 hours.Conclusions (1) The expression of CD25 and CD69 on T lymphocytes was inhibited by simvastatin in vitro, and the higher the simvastatin concentration, the more serious inhibition of the expression of CD25 and CD69 on T lymphocyte surfaces. The inhibition was the most serious after 24 hours or 48 hours; (2) The expression and production of IL-2, IL-6 and TNF-αwas inhibited by simvastatin in vitro, and the higher the simvastatin concentration, the more serious inhibition of IL-2, IL-6 and TNF-αexpression and production. The inhibition was the most serious after 24 hours or 48 hours; (3) The transpotion of NF-κB into the nuclear was inhibited by simvastatin in vitro, and the higher the simvastatin concentration, the more serious inhibition of the transportion of NF-κB into the nuclear. The inhibition was the most serious after 24 hours or 48 hours.So simvastatin had the effect of anti-inflammatory on T lymphocyte by inhibiting the transportion of NF-κB into the nuclear.