The Influence Of ?-spinasterol On The Expression Of P-glycoprotein And PepT1 On The Small Intestine Of Mice

Objective:Based on Traditional Chinese Medicine medicinal guide theory and combined with the similar method of pervious relevant laboratory studies, we choose a-spinasterol extracted from Platycodon as research material and Tylosin tartrate as substrate to research the effect of ?-spinasterol on the intestine transporters P-gp and PepT1 by immunohistochemical staining, fluorescent quantitative PCR and western blotting firstly. And then the mechanism of Platycodon latycodon grandiflomm's guide actiong on absorption is discussed preliminarily, which can provide a basis for further study and can also verify the science of Traditional Chinese Medicine medicinal guide theoryMethod:144 mice were equally divided into four treatment groups:a-spinasterol group, tylenol tartrate group, mixed (?-spinasterol and Tylosin tartrate) group, and normal saline group which was used as a control group. The formulations were administered by oral gavage with a single dose of 0.2 mL·10g-1 BW depending on weight of mice. The small intestine samples of the dissected mice were excised at predetermined time intervals (1,3, 5 days). The expression of P-gp and PepT1 on the mice intestines was detected by immunohistochemical staining, fluorescent quantitative PCR and western blotting at 1,3 and 5 day respectively.Results:The results showed that the protein expression of P-gp was distinctly reduced in a-spinasterol and mixed groups, but increased in tylosin group compared with normal saline group. However, the results of PepT1 was opposite. The difference was more and more significant accompany with time passed(P<0.01). In summary, ?-spinasterol had an obvious impact on intestine transporters, including inhibiting P-gp expression and improving PepT1 expression. From the results of immunohistochemical,the change of P-gp in the duodenum is the most obvious, but the change of PepT1 in jejunum is significantly higher than the duodenum and ileum. In the results of PCR and WB. the expression of P-gp in each bowel is gradually increasing from proximal to distal. At the same time, the expression of PepT1 is the most in duodenum and has a little difference in the jejunum and ileum.Conclusions:In summary, ?-spinasterol had an obvious impact on intestine transporters, including inhibiting P-gp expression and improving PepT1 expression, tylenol tartrate could inhibiting PepTl expression and improving P-gp expression. One of the mechanisms of Platycodon latycodon grandiflomm's guide actiong on absorption is to effect on the transporters of intestine. A beneficial DDI between a-spinasterol and tylosin occurs when the two drugs are co-administered. The two targets of the DDI are P-gp and PepTl located in the small intestine.