Abstract
Introduction It has been reported that the traditional Chinese medicine Huangqin-Tang decoction (HQT) has a protective effect on the epithelial barrier function of ulcerative colitis, but its mechanism has not been fully clarified. This study intends to explore the protective mechanism of HQT in regulating microRNA (miRNA) for the first time. Methods Based on the Balb/c mice ulcerative colitis model, the mice were given a gavage of 0.1 mL/10 g HQT every day for 7 days; on the 8th day, the colon of the mice was dissected, the length of the colon for the mice was measured, and the score was given based on this. Analysis of colonic mucosal injury was conducted by hematoxylin-eosin staining. Then, the differential miRNA was screened and sequenced in colon tissue using the HiSeq platform. And the differential miR-185-3p gene was verified by RT-PCR. Finally, the effects of HQT on miR-185-3p, occludin protein expression, and transepithelial electrical resistance (TEER) value were observed in combination with the CaCo2 intestinal epithelial cell model. Results HQT treatment can alleviate the shortening of colon length and reverse the intestinal mucosal injury. miRNA sequencing of colonic tissue showed that miR-185-3p was significantly downregulated in the model group, while HQT could upregulate miR-185-3p, thereby affecting the myosin light chain kinase (MLCK)/myosin light chain phosphorylation (p-MLC) pathway and leading to increased expression of occludin protein, which ultimately protected the intestinal epithelial barrier function. Conclusion HQT can protect colon epithelial barrier function by regulating miR-185-3p.
Highlights
It has been reported that the traditional Chinese medicine Huangqin-Tang decoction (HQT) has a protective effect on the epithelial barrier function of ulcerative colitis, but its mechanism has not been fully clarified. is study intends to explore the protective mechanism of HQT in regulating microRNA for the first time
Studies have shown that increased intestinal mucosal permeability may be an early event in the onset of ulcerative colitis (UC), and patients with US at rest develop increased intestinal permeability and associated intestinal symptoms even though no significant abnormalities are observed under endoscopy [1,2,3]. erefore, improving patients’ intestinal epithelial barrier function is considered to be the key to effective treatment of UC. e traditional Chinese medicine HQT is a classic prescription for treating dysentery, and it was first recorded in the Treatise on Febrile Diseases by Zhang Zhongjing: the combination of sun disease and Shaoyang disease with diarrhea should be treated with Huangqin-Tang decoction
According to the method reported in our literature, raw medicinal materials including Radix Scutellariae, Radix Paeoniae Alba, Glycyrrhizae Radix, and Fructus Jujubae were mixed according to the ratio of 9 : 6:6 : 49 [12]. e raw medicinal materials were soaked in 10 times the volume of distilled water for 30 min, and decocted them at 100°C for 30 min. en, the extract was collected from the decocted water. 10 times the volume of distilled water was added to the herb residue, and a decoction for it of 30 min was needed. e extract needed to be collected, and the two extractive solutions needed to be combined, concentrated, and freeze-dried
Summary
Intestinal mucosal barrier injury is an important cause of ulcerative colitis (UC), while an injured intestinal mucosal barrier can cause increased intestinal permeability, allowing antigens of bacterial or food origin in the intestinal lumen to enter the mucosa, triggering an immune response in the intestine and an outbreak of an uncontrollable cascade of inflammatory signals. Erefore, improving patients’ intestinal epithelial barrier function is considered to be the key to effective treatment of UC. Baicalin can inhibit NF-KB and regulate Treg/ 17 cell balance in the colon tissue of experimental animals so as to exert anti-inflammatory and immunomodulatory effects [7, 8]. HQT can effectively alleviate experimental ulcerative colitis induced by dextran sulfate sodium (DSS) and 2,4,6-trinitrobenzene sulfonic acid (TNBS) [10, 11]. Animal studies show that HQT can inhibit the pathway (RasPI3K-Akt-HIF-1a and NF-kB) in colon tissue from playing an anti-inflammatory role [12]. MiR-210/miR-155 can regulate the expression of cytokine IFN-c/transcription factor Hif1a/IL-2, affecting the differentiation and function of intestinal adaptive immune cells [18,19,20]. We plan to use colon mucosa tissue to sequence and analyze miRNA and clarify the miRNA that HQT can regulate, so as to provide a scientific basis for improving the therapeutic mechanism of HQT
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