Abstract：
BACKGROUND: Portulacae Herba and Granati Pericarpium pair (PGP) is a traditional Chinese herbal medicine treatment for colitis, clinically demonstrating a relatively favorable effect on relieving diarrhea and abnormal stools. However, the underlying mechanism remain uncertain.
OBJECTIVE: The present study intends to evaluate the efficacy of PGP in treating colitis in mice and investigate its underlying mechanism.
METHODS: The protective effect of PGP against colitis was determined by monitoring body weight, colon length, colon weight, and survival rate in mice. Colonic inflammation was assessed by serum cytokine levels, colonic H&E staining, and local neutrophil infiltration. The reversal of intestinal epithelial barrier damage by PGP was subsequently analyzed with Western blot and histological staining. Furthermore, RNA-seq analysis and molecular docking were performed to identify potential pathways recruited by PGP. Following the hints of the transcriptomic results, the role of PGP through the IL-6/STAT3/SOCS3 pathway in DSS-induced colitis mice was verified by Western blot.
RESULTS: DSS-induced colitis in mice was significantly curbed by PGP treatment. PGP treatment significantly mitigated DSS-induced colitis in mice, as evidenced by improvements in body weight, DAI severity, survival rate, and inflammatory cytokines levels in serum and colon. Moreover, PGP treatment up-regulated the level of Slc26a3, thereby increasing the expressions of the tight junction/adherens junction proteins ZO-1, occludin and E-cadherin in the colon. RNA-seq analysis revealed that PGP inhibits the IL-6/STAT3/SOCS3 pathway at the transcriptional level. Molecular docking indicated that the major components of PGP could bind tightly to the proteins of IL-6 and SOCS3. Meanwhile, the result of Western blot revealed that the IL-6/STAT3/SOCS3 pathway was inhibited at the protein level after PGP administration.
CONCLUSIONS: PGP could alleviate colonic inflammation and reverse damage to the intestinal epithelial barrier in DSS-induced colitis mice. The underlying mechanism involves the inhibition of the IL-6/STAT3/SOCS3 pathway.
OBJECTIVE: The present study intends to evaluate the efficacy of PGP in treating colitis in mice and investigate its underlying mechanism.
METHODS: The protective effect of PGP against colitis was determined by monitoring body weight, colon length, colon weight, and survival rate in mice. Colonic inflammation was assessed by serum cytokine levels, colonic H&E staining, and local neutrophil infiltration. The reversal of intestinal epithelial barrier damage by PGP was subsequently analyzed with Western blot and histological staining. Furthermore, RNA-seq analysis and molecular docking were performed to identify potential pathways recruited by PGP. Following the hints of the transcriptomic results, the role of PGP through the IL-6/STAT3/SOCS3 pathway in DSS-induced colitis mice was verified by Western blot.
RESULTS: DSS-induced colitis in mice was significantly curbed by PGP treatment. PGP treatment significantly mitigated DSS-induced colitis in mice, as evidenced by improvements in body weight, DAI severity, survival rate, and inflammatory cytokines levels in serum and colon. Moreover, PGP treatment up-regulated the level of Slc26a3, thereby increasing the expressions of the tight junction/adherens junction proteins ZO-1, occludin and E-cadherin in the colon. RNA-seq analysis revealed that PGP inhibits the IL-6/STAT3/SOCS3 pathway at the transcriptional level. Molecular docking indicated that the major components of PGP could bind tightly to the proteins of IL-6 and SOCS3. Meanwhile, the result of Western blot revealed that the IL-6/STAT3/SOCS3 pathway was inhibited at the protein level after PGP administration.
CONCLUSIONS: PGP could alleviate colonic inflammation and reverse damage to the intestinal epithelial barrier in DSS-induced colitis mice. The underlying mechanism involves the inhibition of the IL-6/STAT3/SOCS3 pathway.
摘要：
背景：马齿轮轴和葛根对（PGP）是治疗结肠炎的传统中草药，在临床上证明对缓解腹泻和大便异常有相对有利的效果。然而,潜在的机制仍然不确定。
目的：本研究旨在评价PGP治疗小鼠结肠炎的疗效及其机制。
方法：通过监测体重来确定PGP对结肠炎的保护作用，结肠长度,结肠重量,和小鼠的存活率。通过血清细胞因子水平评估结肠炎症，结肠H&E染色,局部中性粒细胞浸润。随后用Western印迹和组织学染色分析PGP对肠上皮屏障损伤的逆转。此外，进行RNA-seq分析和分子对接以鉴定PGP募集的潜在途径。根据转录组结果的提示，通过Westernblot验证了PGP通过IL-6/STAT3/SOCS3途径在DSS诱导的结肠炎小鼠中的作用。
结果：PGP治疗可显著抑制DSS诱导的小鼠结肠炎。PGP治疗可显着减轻DSS诱导的小鼠结肠炎，体重的改善证明了这一点，DAI严重性,存活率,血清和结肠中的炎性细胞因子水平。此外，PGP处理上调了Slc26a3的水平，从而增加了结肠中紧密连接/粘附性连接蛋白ZO-1，闭塞蛋白和E-钙黏着蛋白的表达。RNA-seq分析显示PGP在转录水平上抑制IL-6/STAT3/SOCS3途径。分子对接表明PGP的主要成分可以与IL-6和SOCS3蛋白紧密结合。同时,Westernblot结果显示，PGP给药后，IL-6/STAT3/SOCS3途径在蛋白水平上受到抑制。
结论：PGP能减轻DSS诱导的结肠炎小鼠结肠炎症，逆转肠上皮屏障的损伤。潜在的机制涉及IL-6/STAT3/SOCS3途径的抑制。
目的：本研究旨在评价PGP治疗小鼠结肠炎的疗效及其机制。
方法：通过监测体重来确定PGP对结肠炎的保护作用，结肠长度,结肠重量,和小鼠的存活率。通过血清细胞因子水平评估结肠炎症，结肠H&E染色,局部中性粒细胞浸润。随后用Western印迹和组织学染色分析PGP对肠上皮屏障损伤的逆转。此外，进行RNA-seq分析和分子对接以鉴定PGP募集的潜在途径。根据转录组结果的提示，通过Westernblot验证了PGP通过IL-6/STAT3/SOCS3途径在DSS诱导的结肠炎小鼠中的作用。
结果：PGP治疗可显著抑制DSS诱导的小鼠结肠炎。PGP治疗可显着减轻DSS诱导的小鼠结肠炎，体重的改善证明了这一点，DAI严重性,存活率,血清和结肠中的炎性细胞因子水平。此外，PGP处理上调了Slc26a3的水平，从而增加了结肠中紧密连接/粘附性连接蛋白ZO-1，闭塞蛋白和E-钙黏着蛋白的表达。RNA-seq分析显示PGP在转录水平上抑制IL-6/STAT3/SOCS3途径。分子对接表明PGP的主要成分可以与IL-6和SOCS3蛋白紧密结合。同时,Westernblot结果显示，PGP给药后，IL-6/STAT3/SOCS3途径在蛋白水平上受到抑制。
结论：PGP能减轻DSS诱导的结肠炎小鼠结肠炎症，逆转肠上皮屏障的损伤。潜在的机制涉及IL-6/STAT3/SOCS3途径的抑制。
