Effect Of Macleaya Cordata Alkaloids On The Proliferation Of Intestinal Porcine Epithelial Cells(IPEC-1) And The Expression Of Gastrin

In the present study, the author has investigated the effect of Macleaya Cordata alkaloids on the proliferation of intestinal porcine epithelial cells (IPEC-1) and the expression of gastrin in IPEC-1and HepG2cells.Firstly,to study the effects of sanguinarine, chelerythrine, Protopine and allocryptopine from Macleaya cordata on intestinal porcine epithelial cells(IPEC-1) proliferation at different concentrations and different periods. IPEC-1cells were pre-incubated for24hours, and then exposure to different concentrations (4,2,1,0.5,0.25,0.1,0.05,0.025,0.0125and0.00625μg/mL of sanguinarine or chelerythrine,and100,50,25,12.5,6.25,3.125and1.5625μg/mL of protopine or allocryptopine; each treatment had8replicates) for additional12or24hours, respectively, cell viability was measured by MTT assay. The results showed as follows:after the IPEC-1cells were treated by0.25to4μg/mL of sanguinarine,0.25to4μg/mL of chelerythrine,100μg/ml of protopine or100μg/mL of allocryptopine for12hours, the proliferation of cells was significantly inhibited (P<0.05); whereas0.00625to0.1μg/mL of sanguinarine,0.00625to0.1μg/mL of chelerythrine,3.125to25μg/mL of protopine or1.5625to25μg/mL of allocryptopine significantly promoted IPEC-1proliferation (P<0.05), however, with Macleaya cordata alkaloids concentrations decreasing, the effects had slowly dimnished; when extended the treatment time to24hours, only0.1μg/mL of chelerythrine,25μg/mL of allocryptopine and25μg/mL of protopine showed a significant enhance effect on the IPEC-11proliferation (P<0.05), there were no significant differences between the other experimental groups and control group (P>0.05). The results indicated that the Macleaya cordata alkaloids can promote the proliferation of IPEC-1under certain concentration; it speculates that macleaya cordata alkaloids may improve the intestinal digestive, absorptive, immune and barrier function through this way, and then improve the animal growth performance.Secondly, we studied the effects of Macleaya Cordata alkaloids including sanguinarine, chelerythrine, protopine and allocryptopine on the expression of gastrin in IPEC-1and an liver cancer cell line HepG2, as well the effects of gastrin on the proliferation of these two cell lines. IPEC-1and HepG2cells were treated to different concentrations of gastrin (10-5～10-10mol/L) for12h respectively, cell viability was measured by MTT assay. The result indicated that after the cells were treated using10-7～10-10mol/L of gastrin, the growth of cells was promoted significantly (P<0.05) on both cells, whereas treated with10-5～10-6mol/L of gastrin showed no significant effects (P>0.05). After IPEC-1or HepG2cells were treated using the four Macleaya Cordata alkaloids, the expression levels of gastrin and gastrin receptor were meatured by ELISA assay. The result showed that the levels of gastrin and gastrin receptor were increased significantly (P<0.05) in IPEC-1cells, however there was no significant difference (P>0.05) or inhibited significantly (P<0.05) in HepG2cells.Furthermore, Western Blotting assay was used to measure the expression levels of gastrin and gastrin receptor in HepG2cells after treatment with alkaloids. The results indicated that the expression levels of gastrin and gastrin receptor were increasing inhibited significantly (P<0.05) with dose dependence. These results were consistent with those obtained from the ELISA assay.In summary, Macleaya Cordata alkaloids may promote the proliferation of normal cells of the gastrointestinal tract and could inhibit the growth of tumor cells through the regulation of gastrin.