ABSTRACT
ObjectiveTo observe the effect of Shenling Baizhusan on the intestinal inflammatory reaction in the rat model of Crohn's disease （CD） and study its relationship with p38 mitogen-activated protein kinase （MAPK） signaling pathway， so as to provide an experimental and theoretical basis for the clinical application of this prescription. MethodA total of 72 SD rats （36 males and 36 females） were randomized into a normal group （n=12） and a modeling group （n=60）. The rats in the modeling group were treated with 2，4，6-trinitrobenzene sulfonic acid （TNBS， 3 mL·kg-1） and then randomized into model， mesalazine （0.21 g·kg-1·d-1）， and low-， medium-， and high-dose （5.88， 11.76， 23.59 g·kg-1·d-1， respectively） Shenling Baizhusan groups. The rats in the drug intervention groups were administrated with corresponding agents by gavage for 14 days， and those in the normal and model groups with an equal volume of distilled water. The disease activity index （DAI） score of inflammatory bowel disease （IBD） and the colon mucosal damage index （CMDI） score of rats in each group were assessed after gavage. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the colon， and enzyme-linked immunosorbent assay （ELISA） to measure the levels of tumor necrosis factor alpha （TNF-α）， interleukin-1 （IL-1）， and interleukin-6 （IL-6） in the serum. Western blotting was employed to determine the protein levels of p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， p65 nuclear factor （NF）-κB， and phosphorylated-p65 NF-κB （p-NF-κB p65） in the colon tissue. Quantitative real-time polymerase chain reaction was conducted to determine the miRNA levels of p38 MAPK and NF-κB p65 in the colon tissue. ResultThe model group had higher DAI and CMDI scores than the normal group （P<0.01） and showed damaged epithelial cells in the colon mucosa， disarrangement of glands， damaged simple tubular glands， local necrosis， infiltration of a large number of inflammatory cells and lymphocytes in each layer， and presence of ulceration. Compared with the normal group， the model group showed elevated levels of TNF-α， IL-1， and IL-6 in the serum （P<0.01） and up-regulated protein levels of p-p38 MAPK and p-NF-κB p65 and miRNA level of p38 MAPK in the colon tissue （P<0.01）. Compared with the model group， mesalazine and high- and medium-dose Shenling Baizhusan decreased the DAI and CMDI scores （P<0.05， P<0.01）， repaired the mucosal epithelium of the colon tissue， increased the glands and goblet cells， lowered the levels of TNF-α， IL-1， and IL-6 in the serum （P<0.05， P<0.01）， and down-regulated the protein levels of p-p38 MAPK and p-NF-κB p65 and the miRNA level of p38 MAP in the colon mucosa （P<0.01， P<0.05）. ConclusionShenling Baizhusan can reduce intestinal inflammation of CD rats and promote the repair of colon mucosa by down-regulating the protein levels of p-p38 MAPK and pNF-κB p65 and the miRNA level of p38 MAPK to inhibit the p38 MAPK pathway.
ABSTRACT
The most common manifestations of inflammatory bowel disease （IBD） are Crohn's disease and ulcerative colitis， and the global incidence of IBD is on the rise. Traditional Chinese medicine （TCM） is advantageous in the treatment of IBD. IBD， with TCM names based on clinical symptoms， mostly belongs to recurrent dysentery， long dysentery， diarrhea， dysentery， bowel， and other categories. In TCM pathogenesis of IBD， spleen deficiency and exuberant dampness predominate in the whole course of the disease. Since the lung is associated with the large intestine and the lung Qi and spleen Qi are interconnected， the lung Qi and spleen Qi are deficient and the dampness and heat accumulate internally， which caused collateral obstruction by stagnant blood and the development of IBD. From the perspective of "associating lung with large intestine"，it is believed that the main mechanism of IBD is the Qi imbalance and abnormal metabolism of fluids in the lung and the intestine，and the nutrient-Yin injury of the lung and the intestine. According to the chronic， recurrent， and diffuse pathogenesis characteristics and main clinical manifestations of IBD， IBD is closely related to the lung and the intestine. In terms of therapeutic principles， IBD can be treated by tonifying the spleen and replenishing the lung， which highlights the treatment of the intestine from the lung. To be specific， in time of tonifying the spleen and removing dampness， the intestine is regulated by tonifying the spleen and replenishing the lung. Shenling Baizhusan， a commonly used classical prescription for IBD， is mainly potent in replenishing Qi， invigorating the spleen， draining dampness， checking diarrhea， and especially "reinforcing earth to generate metal". It can enhance the function of the lung through "reinforcing earth to generate metal"， which in turn regulates the intestine and promote the improvement of IBD. The present study clarified the mechanism of Shenling Baizhusan in regulating the intestine by tonifying the spleen and replenishing the lung. On the basis of modern research， its therapeutic effect on IBD was achieved through multiple links， such as regulation of the level of inflammatory factors， immunoregulation， barrier function improvement via mucosal repair， and intestinal flora. The findings of this study are expected to provide new ideas for the regulation of the lung-spleen-large intestine axis in the syndrome differentiation and treatment of IBD and subsequent experimental research.
ABSTRACT
Hoxa3-Pax1/Pax9-Eya1-Six1/4 regulatory pathway seems to be operating during forming the bilateral parathyroid/thymus common primordial in early embryonic development.The specification of the parathyroid domain in the parathyroid/thymus primordial is regulated through a Shh-Tbx1-Gcm2 pathway.Gcm2 also may play roles in later steps of parathyroid development,including CaSR and PTH gene expression.MafB and Gcm2 interact with each other and synergistically activate PTH transcription.Genetic basis and the etiology of some hypoparathyroid disorders in man are involved defects in transcription factors that include GCMB,GATA3,Tbxl,SOX3 and GNA11.This marker expression in thymus and parathyroid primordium includes HoxA3,Pax1,Eya1,and Six1;and expression of parathyroid cell-like cells includes Gcm2,CaSR,and PTH.These expressions may serve as markers of stem cell differentiation into parathyroid cell-like cells.