| 张一,潘晓艳,王秀苇,陈丽霞,谢烨婷,陈坚翱,俞蕾敏.炒白术对肠易激综合征小鼠血管内皮功能及血清Slit2表达的影响[J].浙江中西医结合杂志,2024,34(9): |
| 炒白术对肠易激综合征小鼠血管内皮功能及血清Slit2表达的影响 |
| Effects of fried Atractylodes macrocephala on vascular endothelial function and serum Slit2 expression in mice with irritable bowel syndrome |
| 投稿时间：2023-12-04 修订日期：2024-08-04 |
| DOI： |
| 中文关键词: 炒白术 肠易激综合征 血管内皮功能 肠黏膜屏障 Slit2 |
| 英文关键词:Stir-fried Atractylodes irritable bowel syndrome endothelial function intestinal mucosal barrier Slit2 |
| 基金项目:浙江省中医药科技计划项目(No.2023ZL101) |
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| 中文摘要: |
| 目的 探讨炒白术通过神经生长导向因子2（Slit2）表达改善肠易激综合征（IBS）小鼠肠道血管内皮功能的作用机制。方法 共使用50只正常雌性C57BL/6小鼠，随机选取10只作为对照组，其余采用束缚应激建立IBS小鼠模型，并随机分为模型组、低剂量白术组（0.05 g/mL)、中剂量白术组(0.1 g/mL)、高剂量白术组(0.5 g/mL)，各10只。造模结束10天后，低、中、高剂量白术组予不同剂量炒白术煎剂灌胃治疗，模型组等量生理盐水灌胃7 d，造模结束和治疗结束后利分别用结直肠扩张-腹壁撤回反射（CRD-AWR）法评估小鼠内脏敏感性。酶联免疫吸附试验(ELISA)检测小鼠血清Slit2、白细胞介素1β（IL-1β）、肿瘤坏死因子α（TNF-α）、诱导型一氧化氮合成酶（iNOS）、内皮素1（ET-1）含量，硝酸还原酶法检测小鼠血清一氧化氮（NO）含量，蛋白质印记（Western blot）检测小鼠肠道封闭蛋白1（claudin1）、紧密连接蛋白（occludin）含量。结果 与对照组比较，模型组小鼠腹部撤回反射（AWR）评分在直肠扩张压力为40 mmHg[(2.31±0.49)分比(1.54±0.44)分]和60 mmHg[(3.43±0.55)分比(1.54±0.44)分]时显著升高（P＜0.05或P＜0.01），血清Slit2含量下降[(666.40±152.20)pg/mL比（923.10±146.20）pg/mL，P＜0.01]，IL-1β[（80.31±6.87）pg/mL比（41.95±7.64）pg/mL，P＜0.01]、TNF-α[（81.17±13.62）pg/mL比（34.33±8.31）pg/mL，P＜0.01]、iNOS[(17.70±1.85)ng/mL比(9.22±2.07)ng/mL，P＜0.01]、ET-1[(132.10±9.25)pg/mL比（89.52±6.85）pg/mL，P＜0.01]、NO含量 [(61.69±8.30)pg/mL比(31.02±6.24)pg/mL，P＜0.01]均显著升高，肠道occludin[（0.89±0.09）比（1.08±0.10），P＜0.05]、claudin1含量 [(0.95±0.12)比(1.23±0.10)，P＜0.01]明显下降。与模型组比较，高剂量白术组AWR评分在直肠扩张压力为40 mmHg[(1.64±0.41)分比(2.31±0.49)分]和60 mmHg[(2.76±0.50)分比(3.43±0.55)分]时均明显下降(P＜0.05)，血清Slit2含量显著升高[(943.80±182.20)pg/mL比(666.40±152.20)pg/mL，P＜0.01]，IL-1β[(64.81±8.53)pg/mL比(80.31±6.87)pg/mL，P＜0.01]、TNF-α[(53.36±10.40)pg/mL比(81.17±13.62)pg/mL，P＜0.01]、iNOS[（11.59±1.81）ng/mL比(17.70±1.85)ng/mL，P＜0.01]、 ET-1[(114.20±7.33)pg/mL比(132.10±9.25)pg/mL，P＜0.01]和NO含量[(114.20±7.34)pg/mL比(132.10±9.25)pg/mL，P＜0.01]均显著下降，肠道claudin1蛋白[(1.18±0.13)比(0.95±0.12)，P＜0.05]和occludin蛋白含量 [(1.04±0.10）比（0.89±0.09)，P＜0.05]均显著升高。结论 炒白术可能通过Slit2蛋白改善IBS小鼠内脏高敏感及肠屏障功能。 |
| 英文摘要: |
| Abstract Objective To investigate whether fried Atractylodes can improve intestinal vascular endothelial function in mice with irritable bowel syndrome by affecting Slit2 expression and its possible mechanism. Methods 50 normal female C57BL/6 mice, 10 were randomly selected as control group, and the other mice were treated with binding combined with Restraint stress to establish IBS mouse model, and were randomly divided into model group, low-dose Atractylodes macrocephala group (0.05 g/mL), medium-dose Atractylodes macrocephala group (0.1 g/mL), and high-dose Atractylodes macrocephala group (0.5 g/mL) with 10 mice each. After 10 days of modeling, the treatment group was given fried Atractylodes decoction by intragastric administration, and the model group was given the same amount of normal saline intragastric administration for 7 days. After modeling andthe treatment, colorectal dilatation-abdominal wall retraction reflex (CRD-AWR) method was used to evaluate the visceral sensitivity of mice. The contents of serum Slit2 and IL-1β were detected by ELISA, and the contents of intestinal claudin1 and occludin were detected by Western blot. Result Compared with the control group, the abdominal withdrawal reflex (AWR) scores of the mice in the model group were significantly increased at 40 mmHg (2.31±0.49 vs. 1.54±0.44 points) and 60 mmHg (3.43±0.55 vs. 1.54±0.44 points) (P＜0.05 or P＜0.01), the serum Slit2 content was decreased (666.40±152.20 vs. 923.10±146.20 pg/mL, P＜0.01), and IL-1β (80.31±6.87 vs. 41.95±7.64 pg/mL, P＜0.01), TNF-α (81.17±13.62 vs. 34.33±8.31 pg/mL, P＜0.01), iNOS (17.70±1.85 vs. 9.22±2.07 ng/mL, P＜0.01), ET-1 (132.10±9.25 vs. 89.52±6.85 pg/mL, P＜0.01), and NO content (61.69±8.30 vs. 31.02±6.24 pg/mL，P＜0.01) were significantly increased. The intestinal occludin (0.89±0.09 vs. 1.08±0.10, P＜0.05) and claudin1 (0.95±0.12 vs. 1.23±0.10, P＜0.01) levels were significantly decreased. Compared with the model group, the AWR scores of the high-dose Atractylodes macrocephala group were significantly higher at 40 mmHg (1.64±0.41 vs. 2.31±0.49 points) and 60 mmHg (2.76±0.50 vs. 3.43±0.55 points)(P＜0.05). The levels of serum Slit2 were significantly increased (943.80±182.20 vs. 666.40±152.20 pg/mL, P<0.01), and IL-1β(64.81±8.53 vs. 80.31±6.87 pg/mL, P<0.01) , TNF-α (53.36±10.40 vs. 81.17±13.62 pg/mL, P<0.01), iNOS (11.59±1.81 vs. 17.70±1.85 ng/mL, P<0.01), ET-1 (114.20±7.33 vs. 132.10±9.25 pg/mL, P＜0.01) and NO content (114.20±7.34 vs. 132.10±9.25 pg/mL, P＜0.01) were significantly decreased, and the intestinal claudin1 protein (1.18±0.13 vs. 0.95±0.12, P＜0.05) and intestinal occludin protein (1.04±0.10 vs. 0.89±0.09, P＜0.05) content were significantly increased. Conclusion Stir-fried Atractylodes may improve visceral hypersensitivity and intestinal barrier function of IBS mice through Slit2 protein. |
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