| Objective:Ischemia reperfusion injury(ischemia-reperfusion,injury,IRI)can induce various organelles damage and stress of intestinal epithelial cells especially intestinal epithelial cells,thus affecting the expression of specific protein and the release of cytokines,resulting in intestinal epithelial cell damage or apoptosis.Such as ischemia reperfusion injury,membrane damage,calcium overload,mitochondrial damage,endoplasmic reticulum stress and the activation of related enzymes,which will affect the expression of TXNIP/NLRP3 in cytoplasm was significantly.at the same time the over expression of TXNIP/NLRP3 can promote and regulate the production and release of inflammatory factors such as IL-1 beta and IL-18.This study intends to explore:1.effects and significance of Baicalin on the expression of TXNIP/NLRP3 in different groups,2.effects and mechanism of Baicalin on the expression of TXNIP/NLRP3mRNA in different groups.Methods:Intestinal ischemia-reperfusion injury(IRI)is mainly caused by hypoxia reoxygenation,Uaually hypoxia reoxygenation method was used to establish the model of ischemia-reperfusion injury in rat intestinal epithelial crypt cells(IEC-6).Divide the intestinal epithelial cells(IEC-6)were divided into five groups:normal control group,hypoxia reoxygennation group,hypoxia reoxygennation +Baicalin0.5×10-3mol/Lgroup,hypoxia reoxygennation +Baicalin1.0 X 10-3mol/Lgroup,hypoxia reoxygennation +Baicalin5.0×10-3mol/Lgroup.First,the normal cells were cultured in complete medium,then each group was given conventional culture or hypoxia treatment and adding fresh complete medium or baicalin solution,finally get them into the conventional incubator for 12 hours.Choosing different baicalin concentration to detect cell proliferation by using methylthiazolyl tetrazolium(MTT)assay.Using Western blot to investigate the expression of thioredoxin-interacting protein(TXNIP)and Nod-like receptor protein 3(NLRP3),and adopting Real-time fluorescence quantification(PCR)to analyze the expression of TXNIP mRNA and NLRP3 mRNA.Results:After hypoxia reoxygennation for 12h,comparing with hypoxia reoxygennation group and normal control group、hypoxia reoxygennation +Baicalin group.MTT assay,normal control group absorbance value(group A)0.513±0.006,hypoxia reoxygenation group(group B):0.269±0.004,hypoxia reoxygenation+0.5×10-3 mol/L baicalin group(group C):0.325±0.005,hypoxia reoxygenation＋1.0×10-3 mol/Lbaicalin group(group D):0.382±0.010,hypoxia reoxygenation+5.0x10-3 mol/L baicalin group(group E):0.424±0.006,it reveal that baicalin can significantly reduces the cells apoptosis by hypoxia reoxygenation.TXNIP relative quantity(group A:0.141±0.011,group B:0.420±0.009,group C:0.364±0.010,group D0.294±0.008,group E 0.259±0.007),NLRP3 relative quantity(group A 1±0,group B 1.609±0.026,group C 1.629±0.025,group D 1.615±0.018,group E 1.620±0.015),Control five groups the expression of TXNIP/NLRP3 protein were up-regulated(P<0.05),while the expression of the protein are decreased with the increase of Baicalin concentration(P<0.05),TXNIPmRNA relative quantity(group A 1±0,group B 1.449±0..062,group C 1.460±0.065,group D 1.462±0.082,group E 1.455±0.056),NLRP3mRNA relative quantity(group A 1±0,group B 1.609±0.026,group C1.629±0.025,group D 1.615±0.018,group E 1.620±0.015),it shows the expression of TXNIP/NLRP3mRNA have no significant differences(P>0.05).Conclusion:Baicalin may reduce the expression of TXNIP/NLRP3 after jene to alleviate the injury of intestinal epithelial cells in hypoxia reoxygenation. |
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