Effects of berberine in alleviating DSS induced colonic epithelial cell injury
Objective To investigate the protective effect of berberine on the injury of colon epithelial cells induced by dextran sulfate sodium(DSS).Methods NCM-460 cells were randomly divided into blank group(conventional culture),model group(40 mg·mL-1 DSS)and low-dose group(5 μmol·L-1 berberine),high-dose group(10 μmol·L-1 berberine),siRNA group(10 μmol·L-1 berberine+transfected siRNA plasmid),si-SelS group(10 μmol·L-1 berberine+transfected si-SelS plasmid).The expressions of selenioprotein S(SelS)were detected by Western blot;cell proliferation was detected by cell counting kit-8(CCK-8)and 5-ethynyl-2'-deoxyuridine(EdU)tests;trans epithellal electric resistance(TEER)levels were detected by Milli-cell ERS;superoxide dismutase(SOD)was detected by kit method;apoptosis was detected by flow cytometry.Results The relative expression levels of SelS protein in blank group,model group,high-dose group,siRNA group and si-SelS group were 1.02±0.13,0.42±0.05,0.90±0.08,0.89±0.10 and 0.30±0.03,respectively;the cell optical density at 48 h were 0.85±0.05,0.48±0.04,0.70±0.08,0.68±0.05 and 0.51±0.05,respectively;the EdU positive cell rates were(33.78±2.72)％,(11.90±2.00)％,(25.74±1.94)％,(24.29±1.96)％and(15.17±1.16)％,respectively;TEER values were(100.00±3.64)％,(43.47±4.19)％,(73.28±7.38)％,(76.25±7.68)％and(53.49±4.42)％,respectively;SOD activities were(13.32±0.73),(5.33±0.55),(9.63±1.13),(9.69±0.88)and(6.40±0.57)U·mL-1,respectively;the apoptosis rates were(3.21±0.02)％,(24.59±2.35)％,(10.90±1.09)％,(11.11±1.24)％and(16.73±1.56)％,respectively.The above indexes in the model group were compared with those in the blank group,and those in the high-dose group were compared with those in the model group.The above indexes of si-SelS group were statistically significant compared with those of siRNA group(all P＜0.05).Conclusion Berberine can inhibit oxidative stress and improve DSS induced colon epithelial cell barrier damage by up-regulating SelS.
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