The Influence Of Rosiglitazone On Expression Of Pparγ Of Microglia Activated By Thrombin And Its Mechanism Of Anti-oxidative Stress Reaction

Objective: To observe the effect of precondition of rosiglitazone(RGZ) on the expression of PPARγ, Nrf2, HO-1,NQO1 and γ-GCS in the microglia cells activated by thrombin,and to discuss the mechanism of these reaction in antioxidation. Methods: Microglia cells were obtained from the brain tissues of the newborn rats and were primary cultured in vitro. The microglia cells were isolated in 14 d. The isolated microglia cells were randomly divided into normal control group(control group), thrombin stimulation group(TH group), rosiglitazone intervention group(RGZ+TH group), retinoic acid intervention group(RA+TH group) and Sulforaphane intervention group(SFN+TH group). The expression of PPARγ, Nrf2, HO-1, NQO1 and γ-GCS was observed by immunocytochemistry(ICC), real-time PCR and Western blotting. Results:The immunocytochemistry showed that the number of stained positive cells of PPARγ,Nrf2,HO-1,NQO1 and γ-GCS in TH group, RGZ+TH group, RA+TH group and SFN+TH group were increased remarkably as compared with control group. A significant increase in the PPARγ was observed in the RGZ+TH group compared with the others, and a significant increase in the Nrf2,HO-1,NQO1 and γ-GCS was observed in the SFN+TH group compared with the others. The results of RT-PCR showed that the m RNA expression of PPARγ in RGZ+TH group was increased significantly as compared with control group,TH group, RA+TH group or SFN+TH group(P<0.01),besides, the m RNA expression of Nrf2, HO-1, NQO1 and γ-GCS in SFN+TH group was increased as compared with control group, TH group, RGZ+TH group or RA+TH group(P<0.01). The results of Western blotting showed that the protein levels of PPARγ in RGZ+TH group were significantly increased as compared with control group, TH group,RA+TH group or SFN+TH group(P<0.01). The protein expression of Nrf2, HO-1, NQO1 and γ-GCS in SFN+TH group was increased as compared with the control group, TH group, RGZ+TH group or RA+TH group(P<0.01).Conclusions: Rosiglitazone-pretreatment might increase the expression of PPARγ, Nrf2, HO-1, NQO1 and γ-GCS in the microglia cells activated by the thrombin. By inhibiting and activating the expression of the Nrf2 after retinoic acid-pretreatment, the expression of the downstream gene HO-1, NQO1 and γ-GCS is influenced also. The anti-oxidative stress effects of rosiglitazone might be achieved partly by modulating Nrf2 to control the downstream gene HO-1, NQO1 and γ-GCS.