Jiang Jian-Zhen, Li Wan-Qiu, Kuang Kun-Lin, Jiang Yu-Qing, He Zhao-Xuan, Zhang Li-Juan, Cao Jing-Ya, Wang Dan, Zhang Xin-Yue, Tian Zi-Lei, Zhu Jun, Peng De-Zhong
College of Acupuncture and Tuina, Chengdu University of Traditional Chinese Medicine, Chengdu, China.
Front Endocrinol (Lausanne). 2025 Jun 25;16:1536567. doi: 10.3389/fendo.2025.1536567. eCollection 2025.
BACKGROUND: Diabetic gastroparesis (DGP) is a common complication in the later stage of diabetes mellitus (DM). The aim of this study was to investigate the protective effect of long-term tuina on gastrointestinal (GI) function and the occurrence of DGP in diabetic rats. METHODS: Twenty healthy male SD rats were randomly divided into four groups: NC, DM, DM + GT, and DM + TT. DM was induced with streptozotocin and a high-fat, high-sugar diet for 6 weeks. The DM + TT group received tuina therapy (20 min/session, 5 times/week) for 6 weeks. Weekly random blood glucose, gastric emptying rate, and small intestinal propulsion rate were measured. Hematoxylin and eosin (HE) staining assessed gastric antrum and ileum pathology. Ca-Mg-ATPase and CS activities, and neuronal nitric oxide synthase (nNOS), calmodulin (CaM), and myosin light chain kinase (MLCK) mRNA and protein expressions were evaluated by PCR and Western blot. 5-HT content was measured by ELISA. Piezo2 and 5-HT4R expressions were analyzed by immunofluorescence staining to observe tuina's protective effect on GI function in DM rats. RESULTS: Random blood glucose measurements showed normal levels in the NC group, while other groups remained above 16.7 mmol/L. The DM group exhibited reduced gastric emptying and small intestinal propulsion rates, along with gastric antrum and ileum damage. The DM + TT group showed significant improvements in gastric emptying and intestinal propulsion rates, and reduced tissue damage compared to the DM group. In the DM + TT group, mRNA and protein expressions of CaM and MLCK in antrum tissue, and nNOS, CaM, and MLCK in ileum tissue, were significantly increased. Activities of Ca-Mg-ATPase and CS enzymes in ileum tissue were also elevated, indicating enhanced GI function. Further analysis showed increased mRNA expressions of Piezo2 and 5-HT4R, and protein expressions of Piezo2, 5-HT, and 5-HT4R in the ileum tissues of the DM + TT group. Immunofluorescence intensity of Piezo2 and 5-HT in the ileum was also heightened. These results suggest that tuina's protective effect on GI function is related to the expression of Piezo2 ion channels. CONCLUSIONS: Long-term tuina protects the GI function of DM rats and inhibits the occurrence of DGP, which might be related to the Piezo2/5-HT pathway.
背景：糖尿病胃轻瘫（DGP）是糖尿病（DM）后期的常见并发症。本研究旨在探讨长期推拿对糖尿病大鼠胃肠（GI）功能及DGP发生的保护作用。 方法：将20只健康雄性SD大鼠随机分为四组：正常对照组（NC）、糖尿病组（DM）、糖尿病+普通推拿组（DM + GT）和糖尿病+传统推拿组（DM + TT）。采用链脲佐菌素联合高脂高糖饮食诱导糖尿病6周。DM + TT组接受推拿治疗（每次20分钟，每周5次），共6周。每周测量随机血糖、胃排空率和小肠推进率。苏木精-伊红（HE）染色评估胃窦和回肠病理。通过PCR和Western blot评估钙镁ATP酶（Ca-Mg-ATPase）和胆碱酯酶（CS）活性，以及神经元型一氧化氮合酶（nNOS）、钙调蛋白（CaM）和肌球蛋白轻链激酶（MLCK）的mRNA和蛋白表达。采用ELISA法测定5-羟色胺（5-HT）含量。通过免疫荧光染色分析Piezo2和5-HT4R的表达，以观察推拿对糖尿病大鼠胃肠功能的保护作用。 结果：随机血糖测量显示，NC组血糖水平正常，而其他组血糖水平均高于16.7 mmol/L。DM组胃排空率和小肠推进率降低，同时伴有胃窦和回肠损伤。与DM组相比，DM + TT组胃排空率和小肠推进率显著改善，组织损伤减轻。在DM + TT组中，胃窦组织中CaM和MLCK的mRNA和蛋白表达，以及回肠组织中nNOS、CaM和MLCK的mRNA和蛋白表达均显著增加。回肠组织中Ca-Mg-ATPase和CS酶的活性也升高，表明胃肠功能增强。进一步分析显示，DM + TT组回肠组织中Piezo2和5-HT4R的mRNA表达增加，Piezo2、5-HT和5-HT4R的蛋白表达增加。回肠中Piezo2和5-HT的免疫荧光强度也增强。这些结果表明，推拿对胃肠功能的保护作用与Piezo2离子通道的表达有关。 结论：长期推拿可保护糖尿病大鼠的胃肠功能，抑制DGP的发生，这可能与Piezo2/5-HT途径有关。
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