Clinical Medical College, Chengdu University of Traditional Chinese Medicine, Chengdu, 610075, China.
Department of Traditional Chinese Medicine, the Affiliated Hospital of Southwest Medical University, Luzhou, Sichuang Province, 646000, China.
Chin J Integr Med. 2023 Sep;29(9):809-817. doi: 10.1007/s11655-022-3539-2. Epub 2022 Aug 31.
To explore the possible effects and mechanism of Zhizhu Decoction (ZZD) on the pathophysiology of slow transit constipation (STC).
A total of 54 C57BL/6 mice was randomly divided into the following 6 groups by a random number table, including control, STC model (model), positive control, and low-, medium- and high-doses ZZD treatment groups (5, 10, 20 g/kg, namely L, M-, and H-ZZD, respectively), 9 mice in each group. Following 2-week treatment, intestinal transport rate (ITR) and fecal water content were determined, and blood and colon tissue samples were collected. Hematoxylin-eosin and periodic acid-Schiff staining were performed to evaluate the morphology of colon tissues and calculate the number of goblet cells. To determine intestinal permeability, serum levels of lipopolysaccharide (LPS), low-density lipoprotein (LDL) and mannose were measured using enzyme-linked immunosorbent assay (ELISA). Western blot analysis was carried out to detect the expression levels of intestinal tight junction proteins zona-occludens-1 (ZO-1), claudin-1, occludin and recombinant mucin 2 (MUC2). The mRNA expression levels of inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, IL-4, IL-10 and IL-22 were determined using reverse transcription-quantitative reverse transcription reaction. Colon indexes of oxidative stress were measured by ELISA, and protein expression levels of colon silent information regulator 1/forkhead box O transcription factor 1 (SIRT1/FoxO1) antioxidant signaling pathway were detected by Western blot.
Compared with the model group, ITR and fecal moisture were significantly enhanced in STC mice in the M-ZZD and H-ZZD groups (P<0.01). Additionally, ZZD treatment notably increased the thickness of mucosal and muscular tissue, elevated the number of goblet cells in the colon of STC mice, reduced the secretion levels of LPS, LDL and mannose, and upregulated ZO-1, claudin-1, occludin and MUC2 expressions in the colon in a dose-dependent manner, compared with the model group (P<0.05 or P<0.01). In addition, ZZD significantly attenuated intestinal inflammation and oxidative stress and activated the SIRT1/FoxO1 signaling pathway (P<0.05 or P<0.01).
ZZD exhibited beneficial effects on the intestinal system of STC mice and alleviated intestinal inflammation and oxidative stress via activating SIRT1/FoxO1 antioxidant signaling pathway in the colon.
探讨枳术汤（ZZD）对慢传输型便秘（STC）病理生理学的可能作用及机制。
54 只 C57BL/6 小鼠采用随机数字表法分为 6 组，分别为对照组、STC 模型组（模型组）、阳性对照组及低、中、高剂量 ZZD 治疗组（5、10、20 g/kg，分别为 L、M-和 H-ZZD），每组 9 只。经过 2 周的治疗后，测定肠道传输率（ITR）和粪便含水量，并采集血液和结肠组织样本。采用苏木精-伊红和过碘酸希夫染色评估结肠组织形态，并计算杯状细胞数量。通过酶联免疫吸附试验（ELISA）测定血清中脂多糖（LPS）、低密度脂蛋白（LDL）和甘露糖的水平来评估肠道通透性。采用 Western blot 分析检测肠道紧密连接蛋白 ZO-1、claudin-1、occludin 和重组黏蛋白 2（MUC2）的表达水平。采用逆转录定量聚合酶链反应测定炎症细胞因子肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1β、IL-6、IL-4、IL-10 和 IL-22 的 mRNA 表达水平。通过 ELISA 测定结肠氧化应激指标，通过 Western blot 检测结肠沉默信息调节因子 1/叉头框 O 转录因子 1（SIRT1/FoxO1）抗氧化信号通路的蛋白表达水平。
与模型组相比，M-ZZD 和 H-ZZD 组的 ITR 和粪便水分均显著增加（P<0.01）。此外，与模型组相比，ZZD 治疗可显著增加 STC 小鼠结肠黏膜和肌层的厚度，增加结肠杯状细胞数量，降低 LPS、LDL 和甘露糖的分泌水平，并呈剂量依赖性地上调结肠中 ZO-1、claudin-1、occludin 和 MUC2 的表达（P<0.05 或 P<0.01）。此外，ZZD 可显著减轻肠道炎症和氧化应激，并激活 SIRT1/FoxO1 信号通路（P<0.05 或 P<0.01）。
ZZD 对 STC 小鼠的肠道系统具有有益作用，通过激活结肠中的 SIRT1/FoxO1 抗氧化信号通路减轻肠道炎症和氧化应激。
