Objective:To investigate the expression and interaction of transient receptor potential vanillate subtype 1 (TRPV1) and transient receptor potential cationic channel 8 (TRPM8) proteins in the intestinal tract of mice with acute colitis, and their effects on inflammation and visceral sensation.Methods:A total of 30 male C57BL/6 mice were purchased from Beijing Huafukang Biotechnology, and divided into 3 groups by a random number table method: control group, model group and intervention group, with 10 mice in each group. Mice in model group and intervention group were treated with 30 g/L DSS for 7 days to construct colitis model. Meanwhile, the intervention group was treated with 0.3% WS-12 solution enema every day for 7 consecutive days. At the same time, the vital vitality, hair and body mass changes, fecal characteristics of mice were observed and recorded, the occult blood in feces was measured, and disease activity index (DAI) score was recorded, and the histopathologic score was calculated according to the pathological sections every day. Abdominal wall reflex retreat test was used to detect intestinal sensitivity in each group. The expression levels of colon inflammatory factors including interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF-α) and bradykinin (BK) were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of ZO-1, Occludin, TRPV1, TRPM8, calcitonin gene-related peptide α subunit (CGRP-Gαq) in colon tissue were detected by Western blotting. The mRNA expression levels of inflammatory factors IL-1β and IL-6 in colon tissues and visceral sensitive proteins TRPV1, TRPM8 and Gαq proteins were detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR). The levels of CD4 + T lymphocytes in inflammatory cells were detected by immunohistochemistry. Comparison between the two groups was performed by t test. Results:(1) WS-12 could change the expression levels of intestinal TRPV1, TRPM8 and Gαq proteins: The expression level of intestinal TRPV1 protein in model group was significantly higher than that in control group (2.58±0.25 vs. 1.00±0, t=12.130, P<0.01), and the expression level of Gαq protein in model group was significantly higher than that in control group (2.33±0.51 vs. 1.00±0, t=6.984, P<0.01). The protein expression of TRPM8 in the model group was lower than that in the control group (0.70±0.10 vs. 1.00±0, t=8.001, P<0.01), and the protein expression level of TRPV1 in the intervention group was lower than that in the model group after WS-12 intervention (1.49±0.21 vs. 2.58±0.25, t=11.580, P<0.01), the expression level of Gαq protein in the intervention group was lower than that in the model group (1.34±0.14 vs. 2.33±0.51, t=5.021, P<0.01), and the expression of TRPM8 protein in the intervention group was higher than that in the model group (1.60±0.32 vs. 0.70±0.10, t=6.914, P<0.01). (2) WS-12 could reduce the degree of intestinal inflammation: The colon length of the model group was shorter than that of the control group [(4.01±0.72) cm vs. (7.47±0.55) cm, t=11.960, P<0.01], the DAI score of the model group was higher than that of the control group (3.70±0.48 vs. 0.20±0.42, t=17.260, P<0.01). The histopathologic score of the model group was higher than that of the control group (7.10±0.74 vs. 0.20±0.42, t=25.680, P<0.01), and the expression level of intertissue CD4 + T lymphocytes was increased, and the infiltration was obvious. After WS-12 intervention, colon length was longer [(5.95±0.85) cm vs. (4.01±0.72) cm, t=6.734, P<0.01], DAI score was lower (2.10±0.74 vs. 3.70±0.48, t=5.737, P<0.01) and the histopathologic score of hematoxylin and eosin (HE) was lower (4.80±0.79 vs. 7.10±0.74, t=6.734, P<0.01), and the expression of CD4 + T lymphocytes was lower in the intervention group than in the model group (all P<0.01). The expression levels of IL-1β, IL-6, TNF-α and BK in intestinal tract of model group were significantly higher than those of control group [IL-1β: (107.70±7.86) vs. (23.59±2.26) ng/ml, t=30.190; IL-6: (62.49±6.61) vs. (5.26±1.13) pg/ml, t=27.190; TNF-α: (184.40±11.19) vs. (33.45±6.37) pg/ml, t=37.060; BK: (65.69±7.53) vs. (12.84±4.12) pg/ml, t=19.470, P<0.01]. The above indexes in WS-12 intervention group were lower than those in model group [IL-1β: (42.84±10.45) vs. (107.70±7.86) ng/ml, t=14.230; IL-6: (14.18±5.91) vs. (62.49±6.61) pg/ml, t=17.190; TNF-α: (92.71±3.39) vs. (184.40±11.19) pg/ml, t=8.569; BK: (16.46±3.96) vs. (65.69±7.53) pg/ml, t=18.310, P<0.01]. (3) WS-12 could improve intestinal epithelial permeability: the expression level of ZO-1 and Occludin in the intestinal tract of model group was significantly lower than that of control group (0.58±0.18 vs. 1.00±0, t=10.180; 0.64±0.19 vs. 1.00±0, t=6.466, P<0.01). After WS-12 intervention, the expression of ZO-1 and Occludin in the intervention group was significantly higher than that in the model group (0.88±0.11 vs. 0.58±0.18, t=6.674; 0.82±0.12 vs. 0.64±0.19, t=3.314, P<0.01). (4) The intestinal sensitivity could be improved after the intervention of WS-12: AWR scores in the model group were significantly higher than those in the control group under different pressures (20, 40, 60, 80 mmHg) rectal dilation (0.8±0.4 vs. 0.1±0.3, t=4.200; 1.8±0.4 vs. 0.5±0.5, t=6.091; 2.7±0.5 vs. 1.7±0.4, t=4.629; 3.8±0.4 vs. 2.8±0.6, t=4.160, P<0.01), the intestinal sensitivity of mice was significantly decreased after WS-12 intervention, and AWR score was higher than that of the control group, but it was lower than that of the model group under different pressures (0.4±0.3 vs. 0.8±0.4, t=1.897; 1.1±0.3 vs. 1.8±0.4, t=4.200; 1.9±0.3 vs. 2.7±0.5, t=4.382; 2.9±0.3 vs. 3.8±0.4; t=5.400, P<0.01). Conclusion:There may be a balancing mechanism between TRPV1 and TRPM8 to maintain normal intestinal function. WS-12 can be used to treat acute colitis in mice by activating TRPM8 channel.

目的 探讨血清补体 C1q在炎症性肠病(inflammatory bowel disease,IBD)中的诊断与预后作用.方法 采用回顾性随机直接抽样法,选取 2021 年 1 月至 2022 年 7 月武汉大学人民医院住院患者 274 例,包括 IBD患者 180 例,其中克罗恩病(Crohn's dis-ease,CD)患者 95 例(CD组),溃疡性结肠炎(ulcerative colitis,UC)患者 85 例(UC组),分别采用 CDAI和改良 Mayo评分评估 CD和UC患者的疾病活动度;结直肠癌患者 94 例(结直肠癌组).另选取同期健康体检者 79 名作为正常对照组.比较各组之间的 C1q水平,分析 C1q水平与其他指标的相关性以及 C1q水平与 IBD患者营养状况、器官损伤、电解质水平及凝血功能的关系,用 ROC 曲线评估 C1q预测价值.结果 UC组和 CD组患者血清 C1q 低于正常对照组,结直肠癌组患者血清 C1q 低于 UC 组和 CD 组(P＜0.05);IBD组内,疾病活动度重度组患者血清 C1q 低于缓解期及轻度组;RBC、DBIL、TBIL、白蛋白、肌酐、肾小球滤过率、血钙、血沉、PT、APTT等指标与 C1q明显相关;此外,C1q水平可以评价 IBD 患者营养状况、器官损伤、电解质水平及凝血功能;C1q 对 IBD患者预后有较高的诊断价值.结论 血清 C1q水平对 IBD的病情和预后有潜在的临床价值.

消化道良性狭窄是多种原因引起的消化道管腔缩小的疾病,它发生在食管、贲门、幽门、结直肠以及胆胰管等部位[1-3],引起患者吞咽困难、胸痛、腹痛、腹胀、呕吐、腹部包块、大便异常以及消化道梗阻等症状[4-5],显著降低患者生活质量[6].

Interleukin-1 receptor-associated kinase 1/4 (IRAK1/4) is the main kinase of the Toll-like receptor (TLR)-mediated pathway, considered a new target for treating inflammatory diseases. Studies showed a significant correlation between TLRs and inflammatory responses in ulcerative colitis (UC). Therefore, in this study, after inducing experimental colitis in mice with 3% dextran sulfate sodium (DSS), different concentrations of IRAK1/4 inhibitors were administered intraperitoneally. Then, the disease activity index was assessed, including the degree of pathological damage, by HE staining. Subsequently, while western blotting detected the TLR4/NF-κB pathway and intestinal barrier protein expression (Zonula-1, Occludin, Claudin-1, JAM-A), real-time polymerase chain reaction (RT-PCR) detected the mRNA expression levels of IRAK1/4 and mucin1/2. Furthermore, the expression levels of Zonula-1 and occludin were detected by immunofluorescence, including the plasma FITC-dextran 4000 concentration, to evaluate intestinal barrier permeability. However, ELISA measured the expression of inflammatory factors to reflect intestinal inflammation in mice. Investigations showed that the IRAK 1/4 inhibitor significantly reduced clinical symptoms and pathological DSS-induced colitis damage in mice and then inhibited the cytoplasmic and nuclear translocation of NF-κB p65, including the phosphorylation of IκBα and reduction in downstream inflammatory factor production. Therefore, we established that the IRAK1/4 inhibitor effectively improves colitis induced by DSS, partly by inhibiting the TLR4/NF-κB pathway, reducing inflammation, and maintaining the integrity of the colonic barrier.