The Protective Effect Of P-HSP27on Oxidative Stress Induced By CSE Associate With The Expression Of γ-GCS

【Objective】To detect the expression of HSP27and p-HSP27in lung of passivesmoke rats and AECII dealt with CSE.To investigate the HSP27and p-HSP27expression assicated with the expression of γ-GCS.【Methods】The whole study was divided into two parts: animal research and cellresearch.（1）Animal research:Forty adult male Wistar rats were randomly dividedinto Control group,2month smoking exposed group(2-M-S-G)，4month smokingexposed group(4-M-S-G)and ex-smoking group(exposed to CS for4month and thenquit smoking for1month)respectively. The COPD model was established by passiveexposing with CS.Collecting lung tissue of control and experimental group,theexpression of HSP27and p-HSP27was assayed by Western-blot.（2）Cell research:AECII were cultured. Cigarette smoke extract (CSE) was prepared. At different timesof rat AECII exposed to0%、2.5%、5%、10%CSE, Reverse transcriptase-polymerasechain reaction(RT-PCR) was used to determine the mRNA expression of γ-GCSmRNA. Western blot were used to determine the protein expression of HSP27andP-HSP27.and then AECII treated with5%CSE at different times.The γ-GCS mRNAwas measured by RT-PCR.and the levels of HSP27and p-HSP27expression wasassayed by Western-blot.【Results】（1）Animal research:HSP27protein expression was0.73±0.01、0.77±0.01、0.78±0.03and0.77±0.01in control group、2-M-S-G group、4-M-S-G group and ex-smoking group.The HSP27expression in passive smoke rats had no significant differences with that in controlgroup. The expression of p-HSP27was low in the control group, greatly increased in ratsexposed to2months CS（0.37±0.02VS0.14±0.01，p <0.05）, even higher in rats with4months CS exposure(P <0.05）, and then significantly altered after smoking cessation(P <0.05）, The variationtendency of p-HSP27/HSP27were conformable to that of p-HSP27.（2）Cell research:AECII respectively treated with0%、2.5%、5%、10%CSE at6hand12h,Total HSP27and p-HSP27protein expression was gradually elevated asincreasing concentration of CSE. The expression of total HSP27and p-HSP27proteinwas higher at12h than that at6h.Rat dealt with5%CSE at0h、6h、12h、24h and48h,the expression of HSP27protein was respectively0.12±0.01、0.16±0.02、0.23±0.03、0.29±0.01、0.36±0.01.P-HSP27expression was respectively0.02±0.00、0.06±0.01、0.13±0.02、0.23±0.03、0.23±0.03and the p-HSP27/HSP27was0.17±0.04、0.39±0.07、0.57±0.09、0.80±0.06、0.64±0.09.(The levels expression of total HSP27、p-HSP27and p-HSP27/HSP27were higher at6h and12h than that at0h,lower thanthat at24h,were lower at48h than that at24h,higher than that at6h and12h p<0.05）).AECII exposed to0、2.5%、5%、10%CSE at24h,the expression of γ-GCSmRNA was respectively0.21、0.32、0.44、0.54.The γ-GCS mRNA expression wasrespectively0.27、0.43、0.50、0.7and0.6at0h、6h、12h、24h and48h in AECIIstimulated with concentrations of5%CSE. AECII dealt with5%CSE at0h、6h、12h、24and48h, the γ-GCS protein was up-regulated corresponding to time prolongingand reached the peek at24h. Linear correlation analysis showed the expression ofγ-GCS mRNA and protein and the expression of HSP27、 p-HSP27andp-HSP27/HSP27was positively related.【Conclusion】（1）HSP27may involved the mechanism of the anti-oxidative inpassive smoke rat.（2）The HSP27protect the ACEII treated with CSE fromoxidation.（3）The p-HSP27protect the AECII dealt with CSE from oxidation byup-regulating the expression of γ-GCS...