| 引用本文: |
戚椿林, 廖陈敏, 王文凤, 施敏, 朱哲琴, 魏堰翀, 刘富林, 夏旭婷.枳术丸调节肥大细胞活化及PAR-2/CGRP通路治疗慢传输型便秘脾虚证小鼠的机制研究[J].湖南中医药大学学报,2025,45(8):1413-1419[点击复制] |
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| 枳术丸调节肥大细胞活化及PAR-2/CGRP通路治疗慢传输型便秘脾虚证小鼠的机制研究 |
| 戚椿林,廖陈敏,王文凤,施敏,朱哲琴,魏堰翀,刘富林,夏旭婷 |
| (湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 探讨枳术丸调节肥大细胞活化及蛋白酶激活受体-2(PAR-2)/降钙素基因相关肽（CGRP）通路治疗慢传输型便秘（STC）脾虚证小鼠的机制。方法 30只小鼠以番泻叶灌胃+限制饮食饮水+低纤维饮食方法建立STC脾虚证小鼠模型，并随机均分为模型组（无菌水10 mL/kg灌胃）、莫沙必利组（莫沙必利溶液2.5 mg/kg灌胃）和枳术丸组（枳术丸水煎液9 g/kg灌胃）；另取10只作为正常组（无菌水10 mL/kg灌胃）。干预7 d后，测量小鼠体质量、肠道推进率，HE染色观察结肠黏膜病理，透射电镜观察肥大细胞（MC）形态改变，采用RT-PCR法检测小鼠结肠黏膜类胰蛋白酶（Tryptase）、PAR-2、CGRP m RNA表达，免疫组化检测Tryptase、PAR-2表达，Western blot检测Tryptase、PAR-2蛋白表达，ELISA检测血清、脑组织、结肠黏膜中CGRP含量。结果 与正常组比较，模型组小鼠结肠黏膜水肿、炎症细胞浸润程度、MC形态改变相对加重，炎症病理评分上升（P<0.05），体质量及肠道推进率下降（P<0.01），结肠黏膜Tryptase、PAR-2蛋白表达水平升高（P<0.05,P<0.01）,Tryptase、CGRP、PAR-2 mRNA表达量升高（P<0.01），血清、脑组织、结肠黏膜中CGRP含量升高（P<0.01）。与模型组比较，莫沙必利组与枳术丸组组织水肿、炎症细胞浸润程度、MC形态改变相对减轻，炎症病理评分下降（P<0.05），小鼠体质量和肠道推进率增加（P<0.01）,Tryptase、PAR-2蛋白表达量及Tryptase、CGRP、PAR-2mRNA表达量降低（P<0.05,P<0.01），血清、脑组织、结肠黏膜中CGRP含量降低（P<0.01）。与莫沙必利组比较，枳术丸组小鼠体质量增加（P<0.05）,Tryptase、CGRP、PAR-2 mRNA表达量降低（P<0.05,P<0.01）。结论 枳术丸能改善STC脾虚证小鼠便秘症状，其机制可能与减少MC活化，抑制Tryptase、PAR-2表达，下调CGRP分泌，促进肠道蠕动有关。 |
| 关键词: 慢传输型便秘 枳术丸 肥大细胞 蛋白酶激活受体-2 降钙素基因相关肽 |
| DOI：10.3969/j.issn.1674-070X.2025.08.003 |
| 投稿时间：2025-05-14 |
| 基金项目:湖南省自然科学基金项目(2024JJ7359);湖南省教育厅科学研究项目(23C0164);湖南省教育厅重点项目(23A0292);湖南省中医药科研项目(B2023151,C2022033);中药粉体与创新药物省部共建国家重点实验室培育基地开放基金资助重点项目(24PTKF1001) |
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| Mechanism of Zhizhu Pill on mice with slow transit constipation and spleen deficiency pattern by regulating mast cell activation and PAR-2/CGRP pathway |
| QI Chunlin, LIAO Chenmin, WANG Wenfeng, SHI Min, ZHU Zheqin, WEI Yanchong, LIU Fulin, XIA Xuting |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To investigate the mechanism of Zhizhu Pill on mice with slow transit constipation（STC） and spleen deficiency pattern by regulating mast cell（MC） activation and protease-activated receptor-2（PAR-2）/calcitonin gene-related peptide（CGRP） pathway. Methods Thirty mice were used to establish an STC model with spleen deficiency pattern using a combination of senna leaf gavage, restricted diet and water intake, and a low-fiber diet. These mice were randomly divided into model group（10 mL/kg sterile water gavage, n=10）, mosapride group（2.5 mg/kg mosapride solution gavage, n=10）, and Zhizhu Pill group（9 g/kg Zhizhu Pill decoction gavage, n=10）. Ten additional mice served as the normal group（10 m L/kg sterile water gavage）. After seven consecutive days of intervention, body weight and intestinal propulsion rate of mice were measured. Colonic mucosal pathology was observed via HE staining. MC morphological changes were observed via transmission electron microscopy. The mRNA expressions of tryptase,PAR-2, and CGRP in the colonic mucosa of mice were determined via RT-PCR. Expressions of tryptase and PAR-2 were checked by Immunohistochemistry（IHC）. Protein expressions of tryptase and PAR-2 were measured by Western blot. CGRP content of serum, brain tissue, and colonic mucosa was measured by ELISA. Results Compared with the normal group, mice in the model group demonstrated aggravated colonic mucosal edema, inflammatory cell infiltration, and MC morphological changes accompanied by significantly increased inflammatory pathological points（P＜0.05）, decreased body weight and intestinal propulsion rate（P＜0.01）,elevated protein expressions of tryptase and PAR-2 in colonic mucosa（P＜0.05, P＜0.01）, increased mRNA expressions of tryptase,CGRP, and PAR-2（P＜0.01）, and increased CGRP content in serum, brain tissue, and colonic mucosa（P＜0.01）. Compared with the model group, both mosapride and Zhizhu Pill groups exhibited ameliorated colonic mucosal edema, inflammatory cell infiltration, and MC morphological changes with decreased inflammatory pathological points（P＜0.05）, along with increased body weight and intestinal propulsion rate（P＜0.01）, reduced protein expressions of tryptase and PAR-2, decreased mRNA expressions of tryptase, CGRP, and PAR-2（P＜0.05, P＜0.01）, and reduced CGRP content of serum, brain tissue, and colonic mucosa（P＜0.01）. Compared with the mosapride group, mice in the Zhizhu Pill group showed increased body weight（P＜0.05） and decreased mRNA expressions of tryptase, CGRP,and PAR-2（P＜0.05, P＜0.01）. Conclusion Zhizhu Pill can alleviate constipation symptoms in STC with spleen deficiency pattern. Its mechanism may be related to the suppression of MC activation, inhibition of tryptase and PAR-2 expression, downregulation of CGRP secretion, and promotion of intestinal motility. |
| Key words: slow transit constipation Zhizhu Pill mast cell protease-activated receptor-2 calcitonin gene-related peptide |
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