The Change Of Gr And Hsp90 And Histone Deacetylase 6 After The Use Of Adrenal Cortex Hormone And The Intervention Of Traditional Chinese Medicine

Glucocorticoid is the liposoluble steroid which the adrenal cortex secretes, and it has the anti-inflammatory, antiallergic and immuno-suppressant actions. However, the hypothalamus-pituitary-adrenal cortex is refrained after glucocorticoid is used for a long time, which affects the function of glucocorticoid and which makes it difficult to evacuate the receptor. Glucocorticoid acts on by combining GR and glucocorticoid itself, and it needs heat shock protein 90 as the chaprone. Heat shock protein 90 chaprone's activity is usually adjusted by the reversible acetylation and is controlled by the Histone deacetylase 6 that lies in the cytosol. The experiment is mainly discussing whether the GR and HSP90 and the Histone deacetylase 6 change after the long-time use of GC and administrated You Gui Wan by gavage and the significance of the change.MaterialSD rats; Real-time quantitative PCR amplification; Reverse transcriptase kit; Realtime-pcr kit; Lymphocyte separation medium; DEPC; dexamethasone; You Gui Wan;TRIZOL.MethodForty-five rats, cleaning level, male, four months old, about 200g, were divided equally into 3 groups at random at the time of testing,15 rats in each group.(1) Conrol groupThe animals were daily administrated by intramuscular injection of normal salt solution (NS) 1ml/kg, for 15 days. From the sixteenth day on, the animals administrated NS by gavage 4ml, as well as the intramuscular injection of NS, and lasted another 15 days.(2) Dexamethasone groupThe animals were daily administrated by intramuscular injection of 5mg/ml dexamethasone sodium phosphate solution (1mg/kg), for 15 days. From the sixteenth day on, the doze decreased to half, and lasted another 15 days.(3) Dexamethasone & TCM groupThe animals were daily administrated by intramuscular injection of (5mg/ml) dexamethasone (1mg/kg), for 15 days. From the sixteenth day on, the doze decreased to half, and lasted another 15 days, at the same time, the animals administrated You Gui Wan by gavage, and the doze is 20ml/kg, one time each day, and lasted another 15 days.Draw blood, then separate PBMC, withdraws total RNA, reverse transcription, realtime-pcr.Statistical analysis:SPSS was used to deal with all the data. Analyses of variance(ANOVA) was performed, A value of P<0.05 was considered to indicate a significant difference.ResultsCompared with the control group, the expression level of GR mRNA of the Dexamethasone group decreased clearly, Compared with the control group, the expression level of GR mRNA of the Dexamethasone & TCM group decreased clearly, and the difference is very clear.Compared with the control group, the expression level of HSP90βmRNA of the Dexamethasone group decreased clearly, Compared with the Dexamethasone group, the expression level of HSP90βmRNA of the Dexamethasone & TCM group increased clearly,but is lower than control group.The HDAC6mRNA expression level of the Dexamethasone & TCM group is much higher than the control group and Dexamethasone group.DiscussionHeat shock protein90P has related to the anti-apoposis and medicine resistance. The experiment shows that HSP90βfell down after the use of GC. It is guessed that the anti-apoposis ability dropped, and the mononuclear cell's apoposis increased. There was a slight increase in HSP90βafter the use of You Gui Wan. It is guessed that the anti-apoposis ability enhanced.Hsp90 chaperone activity is regulated by reversible acetylation and controlled by the deacetylase HDAC6. HDAC6 functions as an Hsp90 deacetylase. Inactivation of HDAC6 leads to Hsp90 hyperacetylation and a loss of chaperone activity. In HDAC6-deficient cells, Hsp90-dependent maturation of the glucocorticoid receptor (GR) is compromised.The experiment shows that the rats'peripheral mononu cell's HDCA6 increased after the use of You Gui Wan than the injection of GC and control group. So it is guessed that You Gui Wan has something to do with the regulation of HDCA6, and increased the stability of GR.Conclusion1. The expression of HSP90βfell down after the the long term use of GC, There was a slight increase in HSP90βafter the use of You Gui Wan.2. After the the long term use of GC and then given You Gui Wan, the expression of HDAC6 increased.