黄连-木香介导糖萼重塑保护肠道血管内皮屏障缓解溃疡性结肠炎小鼠肠道炎症OA

This study investigates the potential mechanism underlying the therapeutic effect of Coptidis Rhizoma-Aucklandiae Radix for ulcerative colitis(UC)in mice from glycocalyx remodeling.The mouse model of UC was established with 2.5%dextran sodium sulfate(DSS).Fifty C57BL/6J mice were randomly assigned to five groups:control,model,sulfasalazine(100 mg·kg-1),low-dose(0.64 g·kg-1)Coptidis Rhizoma-Aucklandiae Radix,and high-dose(1.28 g·kg-1)Coptidis Rhizoma-Aucklandiae Radix.Changes in body weight and hematochezia were monitored throughout the experiment,and the disease activity index(DAI)was calculated accordingly.At the end of the experiment,hematoxylin-eosin(HE)staining was performed to evaluate histopathological changes in the colon tissue.Evans blue dye extravasation was employed to assess colonic vascular permeability.RT-qPCR was conducted to quantify the mRNA levels of inflammatory cytokines,markers of intestinal barrier injury,and glycocalyx-related genes in the colon tissue.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the serum levels of shed hyaluronic acid(HA).Immunofluorescence staining was adopted to detect HA-positive signals in the colonic glycocalyx.Transmission electron microscopy(TEM)was utilized to examine the ultrastructural integrity of colonic vascular endothelial cells and the extent of glycocalyx shedding.The results demonstrated that compared with the control group,the model group exhibited significant body weight loss,marked shortening of the colon,elevated DAI,severe hematochezia,pronounced inflammatory cell infiltration,increased colonic vascular permeability,upregulated expression of pro-inflammatory cytokines including tumor necrosis factor(TNF)-α,interleukin(IL)-6,and IL-1β,downregulated expression of tight junction proteins such as Zonula occludens(ZO)-1,Occludin,and Claudin-5,increased expression of vascular cell adhesion molecule(VCAM)-1,altered expression of glycocalyx-modifying enzymes,specifically increased matrix metalloproteinase 1(MMP1)and hyaluronidase 2(Hyal2)and decreased hyaluronan synthase 2(HAS2),along with substantial shedding of endothelial glycocalyx-derived HA into circulation,as evidenced by elevated serum HA concentrations,and severe ultrastructural damage of colonic vascular endothelial cells with marked glycocalyx degradation as revealed by TEM.In comparison with the model group,both the low-and high-dose Coptidis Rhizoma-Aucklandiae Radix groups showed significantly attenuated weight loss and hematochezia,significantly lowered DAI scores,reduced inflammatory infiltration,improved intestinal barrier function,declined levels of TNF-α,IL-6,and IL-1β,restored expression of ZO-1,Occludin,and Claudin-5,reduced VCAM-1 expression,significantly downregulated expression of MMP1 and Hyal2,significantly increased HAS2 expression,suppressed HA shedding,and preserved endothelial glycocalyx structure with less ultrastructural damage observed under TEM.In summary,Coptidis Rhizoma-Aucklandiae Radix ameliorates UC in mice,potentially through mediating glycocalyx remodeling and thereby protecting the integrity of the intestinal vascular endothelial barrier.