Effect of Tongxie Yaofang on p38 MAPK Pathway Associated Protein and Its Target Gene Expression in Colon Tissues of D-IBS Rats with Liver-stagnation and Spleen-deficiency / 中国实验方剂学杂志

ABSTRACT

Objective:To explore the effect of Tongxie Yaofang on p38 mitogen activated protease(p38 MAPK), mitogen and stress protein kinase 1(MSK1), cyclic adenosine effector response element binding protein(CREB)mRNA and protein expression in colon tissue of diarrhea type irritable bowel syndrome (D-IBS) rat model with liver depression and spleen deficiency(GYPX), and interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and the content of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in serum. Method:The 60 SPF Wistar rats were randomly divided into 6 groups, with 10 rats in each group. Except the normal group, rats in the other groups were given by gavage with folium sennae and chronic bondage to establish D-IBS with GYPX for 14 days. The low, medium, and high doses Tongxie Yaofang were administered to Tongxie Yaofang(2.25, 4.5, 9 g·kg-1)gavage respectively. The piveronium bromide group was given piveronium bromide tablets suspension(0.02 g·kg-1)gavage.The normal group group and model group were given the same volume normal saline for 21 days. After the last gavage for 18 hours, the heart blood was collected and the colon tissue was dissected. Real-time PCR was used to observe the expression of p38 MAPK, MSK1 and CREB mRNA in rat colon. Western blot was used to observe the expression of p38 MAPK, MSK1 and CREB protein. ELISA was used to observe the contents of IL-1β, IL-6 and TNF-α in colon. Hydroxylamine was used to observe the T-SOD level in serum, thiobarbituric acid(TBA)was used to observe the MDA content in serum. hematoxyl in-eosin(HE) staining was used to observe the morphological changes of colon tissues. Result:Compared with normal group, the expression of p38 MAPK, MSK1, CREB mRNA and the protein content of p38 MAPK, MSK1 and CREB in the colon tissue of model group rats increased significantly, while the content of IL-1β, IL-6 and TNF-α increased significantly(P<0.05). The level of serum T-SOD decreased significantly, and the content of MDA increased significantly(P<0.05). Compared with the model group, the medium and high dose group of Tongxie Yaofang significantly decreased the expression of p38 MAPK mRNA, content of p38 MAPK, CREB protein and IL-1β, IL-6, TNF-α in colon tissue(P<0.05). The level of serum T-SOD increased significantly, and the content of MDA decreased significantly(P<0.05). High dose group of Tongxie Yaofang can significantly decreased the expression of MSK1, CREB mRNA, content of MSK1 protein(P<0.05). Histopathological observation showed that no significant organic lesions were observed in the colonic morphology of each group of rats, which was consistent with the morphological characteristics of IBS. Conclusion:Tongxie Yaofang has a significant dose-effect relationship in the treatment of D-IBS rats with GYPX in a certain range, which may be related to its increases antioxidant stress and inhibit activation of p38 MAPK signaling pathway and reducing the level of downstream inflammatory factors.