Effects Of Koumine On IPEC-J2 Cells Induced By LPS

Objective:Gelsemium is anti-tumor and sedative,and its main ingredient for pharmacological effects is steroidal alkaloids.Throughout history till today,there always are reports that gelsemium whole grass is used to treat various diseases our country.Koumine is the most abundant alkaloid monomer in the gelsemium whole grass.Porcine jejunal epithelial cells(IPEC-J2)are used as research objects,in this study,to construct small intestime vitro cell inflammation model,and explore the effects of koumine on IPEC-J2 cells induced by Lipopolysaccharides(LPS)under different concentrations by modern molecular biology and other methods,and provide experimental basis for the comprehensive development and utilization of gelsemium.Methods:IPEC-J2 cells were used as the research objects,in this study,and cell viability was detected by MTT method after treated 24h under different koumine concentrations(25,50,100 ? g/mL).After the IPEC-J2 cells were treated with the koumine solution for 1h,the LPS were added to co-react with the cells for 23h.And then,the cell viability of the IPEC-J2 cells,the expression of oxidative damage-related markers,such as SOD(Supweoxide dismutase),CAT(Catalase),MDA(Malondialdehyde)LDH(Lactate dehydrogenase)and so on,were detected by MTT method.The mRNA expressions of TNF-?,IL-6 and IL-1?,and the mRNA expression of ZO-1,Ocdudin,Claudin-1 were detected by q-PCR.Moreover,ELISA was used to detect the content of cytokines TNF-? and IL-6 in cell supernatant.Western blot were used to detect the expression of Zonula Ocdudinludens-1,Ocdudin and Claudin-lin intestinal mechanical barrier.Results:Koumine under the concentrations of 25,50,and 100 ?g/mL could improve the cell viability of IPEC-J2.After the IPEC-J2 cells were pretreated for 1h by koumine under the concentrations of 50,and 100 ? g/mL,to a certain extent,the kouminek could antagonize the decrease of IPEC-J2 activity induced by LPS,relieve the effect of LPS on the IPEC-J2 cells morphology,reduce the contents of MDA,LDH in cells,effectively increase the activity of CAT and SOD,reduce the contents of cytokines TNF-? and IL-6 in cell supernatant,reduce mRNA expressions of TNF-??IL-6?IL-1? in cells,improve the expressions of ZO-1,Ocdudin,Claudin-1mRNA and proteins.Conclusions:Koumine had no toxic effect on IPEC-J2 in certain concentration range,and it could promote the cell viability IPEC-J2,relieve the effect of LPS on the IPEC-J2 cells viability and morphology,improve the anti-inflammatory and antioxidant effects of IPEC-J2 cells,and could reduce the damage of intestinal barrier function caused by LPS,to a certain extent,then maintain the normal intestinal function in animals.