Regulatory effects of the p38 mitogen-activated protein kinase-myosin light chain kinase pathway on the intestinal epithelial mechanical barrier and the mechanism of modified Pulsatilla decoction in the treatment of ulcerative colitis.

OBJECTIVE: To investigate the mechanism of the protective effect of modified Pulsatilla decoction (, MPD) on the mechanical barrier of the ulcerative colitis (UC) intestinal epithelium in vitro and in vivo. METHODS: We established an intestinal epithelial crypt cell line-6 cell barrier injury model by using lipopolysaccharide (LPS). The model was then treated with p38 mitogen-activated protein kinase-myosin light chain kinase (p38MAPK-MLCK) pathway inhibitors, p38MAPK-MLCK pathway silencing genes (si-p38MAPK, si-NF-κB, and si-MLCK), and MPD respectively. Transepithelial electronic resistance (TEER) measurements and permeability assays were performed to assess barrier function. Immunofluorescence staining of tight junctions (TJ) was performed. In in vivo experiment, dextran sodium sulfate-induced colitis rat model was conducted to evaluate the effect of MPD and mesalazine on UC. The rats were scored using the disease activity index based on their clinical symptoms. Transmission electron microscopy and hematoxylin-eosin staining were used to examine morphological changes in UC rats. Western blotting and real-time quantitative polymerase chain reaction were performed to examine the gene and protein expression of significant differential molecules. RESULTS: In in vitro study, LPS-induced intestinal barrier dysfunction was inhibited by p38MAPK-MLCK pathway inhibitors and p38MAPK-MLCK pathway gene silencing. Silencing of p38MAPK-MLCK pathway genes decreased TJ expression. MPD treatment partly restored the LPS-induced decreased in TEER and increase in permeability. MPD increased the gene and protein expression of TJ, while down-regulated the LPS-induced high expression of p-p38MAPK and p-MLC. In UC model rats, MPD could ameliorate body weight loss and disease activity index, relieve colonic pathology, up-regulate TJ expression as well as decrease the expression of p-p38MAPK and p-MLC in UC rat colonic mucosal tissue. CONCLUSIONS: The p38MAPK-MLCK signaling pathway can affect mechanical barrier function and TJ expression in the intestinal epithelium. MPD restores TJ expression and attenuates intestinal epithelial barrier damage by suppressing the p38MAPK-MLCK pathway.

Research progress in regulating intestinal flora structure and repairing intestinal mucosal barrier damage through Tongfu method / 国际中医中药杂志

The intestinal flora and gut barrier function are of great significance for gut function and human health. When the intestinal flora is disrupted and the gut barrier structure is disrupted, it can lead to bacterial translocation, endotoxin influx into the bloodstream, and the production of pro-inflammatory factors, leading to various tissue damage in the body. Tongfu method in TCM can affect the intestinal environment by regulating intestinal permeability and immune response, restoring normal intestinal movement, and regulating the structure and metabolites of intestinal flora, thereby maintaining intestinal homeostasis and body health. The research on regulating intestinal flora and improving intestinal barrier function by Tongfu method can provide reference for further research on the relationship between TCM and intestinal microecology, and provide ideas for clinical treatment.

Effects of Jianpi Mixture on Colonic miRNA Expression Profile in Diarrheal Irritable Bowel Syndrome Rats / 世界科学技术-中医药现代化

Objective To observe the difference in the expression of micro-ribonucleic acid(miRNA)in the colon tissue of irritable bowel syndrome-diarrhea(IBS-D)rats and rats treated with Jianpi Mixture,and predict The miRNA and corresponding genes involved in the treatment of IBS-D with Jianpi Mixture,providing a basis for finding potential therapeutic targets for IBS-D.Methods The rats were randomly divided into a blank group,a model group and a traditional Chinese medicine group,with 8 rats in each group.Both the model group and the Chinese medicine group used acute and chronic stimulation + senna leaf gavage to make the IBS-D model.The Chinese medicine group was gavaged with Jianpi Mixture after model building for 14 days.General condition,body weight and Bristol score were observed and recorded.Hematoxylin-eosin(HE)staining was used to observe the histopathological changes of colon in each group of rats.Transcriptome sequencing of rat colon tissue in each group to screen and map differentially expressed miRNAs.Perform GO(Gene oncology)functional enrichment analysis and KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway enrichment analysis on the target genes corresponding to the screened common miRNAs.Results Compared with the model group,the weight of the rats in the Chinese medicine group increased significantly(P<0.01),and the Bristol score of the feces decreased significantly(P<0.01).The pathological damage of the colon tissue of rats in the Chinese medicine group was significantly reduced.Compared with the blank group,there were 109 differentially expressed genes in the model group,of which 80 were up-regulated and 29 were down-regulated.Compared with the model group,there were 109 differentially expressed genes in the traditional Chinese medicine group,of which 19 were up-regulated and 90 were down-regulated.Mapping the genes of the two comparisons found that 74 miRNAs in the model group were increased and 7 were decreased.The changes of these miRNAs were all reversed after the intervention of traditional Chinese medicine.The GO function enrichment analysis of target genes showed that Jianpi Mixture was mainly involved in synaptic vesicle positioning and transport,cytoplasmic transport,negative regulation of growth and development,and sugar transport after acting on rats.KEGG pathway enrichment analysis showed that MAPK,Wnt,Hippo,TNF,oxygen toxin,cAMP and other pathways were enriched.Conclusion After the intervention of Jianpi Mixture,IBS-D-related miRNA expression profiles have changed significantly;Jianpi Mixture may play a protective role in the treatment of IBS-D by regulating the MAPK signaling pathway and Wnt signaling pathway.Screening the core genes and predicting the miRNAs interacting with them provide a theoretical basis for the study of the pathogenesis of IBS-D and the therapeutic targets of Jianpi Mixture.

Study on the bifidobacterial adhesion to the host tissue in ulcerative colitis and related serum inflammatory factors with Pingkui enema / 国际中医中药杂志

Objective@#This study is designed to investigate the effect and mechanism of Pingkui enema in treatment of ulcerative colitis.@*Methods@#Sixty SD rats at SPF level were randomly divided into 6 groups which were normal group, model group, and the low, medium and high dose groups of Chinese medicine Pingkui enema, and sulfasalazine (SASP) enema group. The normal group and model group were given an equal volume of saline enema after modeling. After continuous administration for 10 days, the histopathological effects were evaluated after extraction, and the serum interleukin-8 (IL-8), interleukin-13 (IL-13), tumor necrosis factor (TNF-α), rat intestinal mucosal bifidobacteria adhesion and adhesin receptor were measured by enzyme-linked immunosorbent assay (ELISA). The real-time fluorescence quantitative PCR (RT-PCR) were used to detect the intestinal bifidobacterial content.@*Results@#Compared with the model group, the content of IL-8 (153.50 ± 8.59 pg/ml, 150.84 ± 5.38 pg/ml vs. 167.13 ± 13.66 pg/ml), TNF-α (330.08 ± 10.02 pg/ml, 287.27 ± 13.89 pg/ml vs. 376.50 ± 19.06 pg/ml) in medium, high dose group of Pingkui enema significantly decreased (P<0.05), while the IL-13 (37.04 ± 3.62 pg/ml, 40.64 ± 5.98 pg/ml vs. 31.57 ± 2.95 pg/ml) significantly increased (P<0.05). The bifidobacteria adhesin (124.84 ± 9.74 ng/L, 149.93 ± 9.57 ng/L, 176.74 ± 11.01 ng/L vs. 101.11 ± 17.18 ng/L) and adhesin receptor (78.44 ± 14.03 ng/L, 99.50 ± 3.63 ng/L, 107.36 ± 5.05 ng/L vs. 60.96 ± 13.89 ng/L) in low-, medium-, high dose group of Pingkui enema significantly increased (P<0.05). The content of bifidobacteria (1 331.4 ± 242.2 copies/μl vs. 490.5 ± 106.0 copies/μl) in the middle dose Pingkui enema group significantly increased (P<0.05).@*Conclusions@#The Pingkui enema can promote the colonization of intestinal epithelial cells by promoting bifidobacteria, increase bifidobacterium adhesin and adhesin receptors, up-regulate the expression of IL-13, and down-regulate the expression of IL-8, TNF-α.

Study on the bifidobacterial adhesion to the host tissue in ulcerative colitis and related serum inflammatory factors with Pingkui enema / 国际中医中药杂志

Objective This study is designed to investigate the effect and mechanism of Pingkui enema in treatment of ulcerative colitis. Methods Sixty SD rats at SPF level were randomly divided into 6 groups which were normal group, model group, and the low, medium and high dose groups of Chinese medicine Pingkui enema, and sulfasalazine (SASP) enema group. The normal group and model group were given an equal volume of saline enema after modeling. After continuous administration for 10 days, the histopathological effects were evaluated after extraction, and the serum interleukin-8 (IL-8), interleukin-13 (IL-13), tumor necrosis factor (TNF-α), rat intestinal mucosal bifidobacteria adhesion and adhesin receptor were measured by enzyme-linked immunosorbent assay (ELISA). The real-time fluorescence quantitative PCR (RT-PCR) were used to detect the intestinal bifidobacterial content. Results Compared with the model group, the content of IL-8 (153.50 ± 8.59 pg/ml, 150.84 ± 5.38 pg/ml vs. 167.13 ± 13.66 pg/ml), TNF-α (330.08 ± 10.02 pg/ml, 287.27 ± 13.89 pg/ml vs. 376.50 ± 19.06 pg/ml) in medium, high dose group of Pingkui enema significantly decreased (P<0.05), while the IL-13 (37.04 ± 3.62 pg/ml, 40.64 ± 5.98 pg/ml vs. 31.57 ± 2.95 pg/ml) significantly increased (P<0.05). The bifidobacteria adhesin (124.84 ± 9.74 ng/L, 149.93 ± 9.57 ng/L, 176.74 ± 11.01 ng/L vs. 101.11 ± 17.18 ng/L) and adhesin receptor (78.44 ± 14.03 ng/L, 99.50 ± 3.63 ng/L, 107.36 ± 5.05 ng/L vs. 60.96 ± 13.89 ng/L) in low-, medium-, high dose group of Pingkui enema significantly increased (P<0.05). The content of bifidobacteria (1 331.4 ± 242.2 copies/μl vs. 490.5 ± 106.0 copies/μl) in the middle dose Pingkui enema group significantly increased (P<0.05). Conclusions The Pingkui enema can promote the colonization of intestinal epithelial cells by promoting bifidobacteria, increase bifidobacterium adhesin and adhesin receptors, up-regulate the expression of IL-13, and down-regulate the expression of IL-8, TNF-α.

Treatment of mammary duct fistula / 中国普通外科杂志

Objective To evaluate the effect of operation on breast duct fishtula. Methods 41 patients with breast duct fitula were subjected to fistulectomy or mastectomy. Results All patients had no re ccurrence after operation from 0.5 to 17 years. The clinical analysis showed that the causes of breast duct fistula were bacterial infection, retracted nipple, tissuration in the middle of nipple and breast duct dialation. Conclusions Fistulectomy or mastectomy is the most effective treatment of breast duct fistula.