湖南中医药大学针灸推拿与康复学院,湖南省长沙市岳麓区学士路300号,410208
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收稿:2024-02-29,
修回:2024-04-13,
纸质出版:2024-07-17
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周竞颖,尹鸿智,刘倩等.电针“内关”“中脘”穴组对功能性消化不良模型大鼠胃动力以及胃窦组织代谢物的影响[J].中医杂志,2024,65(14):1478-1487.
ZHOU Jingying,YIN Hongzhi,LIU Qian,et al.Effects of Electroacupuncture at "Neiguan" (PC6) - "Zhongwan" (RN12) for Rats with Functional Dyspepsia on Gastric Motility and Metabolites of Antral Tissue[J].Journal of Traditional Chinese Medicine,2024,65(14):1478-1487.
周竞颖,尹鸿智,刘倩等.电针“内关”“中脘”穴组对功能性消化不良模型大鼠胃动力以及胃窦组织代谢物的影响[J].中医杂志,2024,65(14):1478-1487. DOI: 10.13288/j.11-2166/r.2024.14.011.
ZHOU Jingying,YIN Hongzhi,LIU Qian,et al.Effects of Electroacupuncture at "Neiguan" (PC6) - "Zhongwan" (RN12) for Rats with Functional Dyspepsia on Gastric Motility and Metabolites of Antral Tissue[J].Journal of Traditional Chinese Medicine,2024,65(14):1478-1487. DOI: 10.13288/j.11-2166/r.2024.14.011.
目的
2
探讨电针“内关”“中脘”穴组治疗功能性消化不良(FD)的可能作用机制。
方法
2
SPF级雄性SD大鼠随机分为模型组、内关组、中脘组、内关-中脘组,每组8只。除正常组外,其余各组大鼠采用0.1%蔗糖碘乙酰胺溶液灌胃联合小平台站立训练建立FD大鼠模型。造模成功后,正常组、模型组大鼠予以捆绑,每天30 min,持续7天;内关组、中脘组、内关-中脘组分别予以电针干预“内关”“中脘”以及“内关”“中脘”穴组,采用连续波,频率2 Hz,电流强度1 mA,每天30 min,持续干预7天。观察各组大鼠一般情况,干预结束后测量各组大鼠体质量、进食量,计算胃排空率;取胃窦组织行HE染色,观察大鼠胃窦组织病理形态学改变;Western Blot法检测胃窦组织中降钙素基因相关肽(CGRP)、胃饥饿素(Ghrelin)蛋白表达;液相色谱-质谱联用(LC-MS)检测胃窦组织代谢物含量,通过相关分析筛选出差异代谢物,利用Metabo Analyst 5. 0和KEGG数据库进行代谢通路分析。
结果
2
光镜下可见各组胃窦组织结构完整,腺体丰富,胃黏膜未见明显炎症、水肿等损伤,均为正常组织。与正常组比较,模型组大鼠体质量、进食量以及胃排空率降低,胃窦组织Ghrelin蛋白表达降低、CGRP蛋白表达升高(
P
<0.01)。与模型组比较,内关组、中脘组、内关-中脘组大鼠体质量、进食量、胃排空率均上升,内关组CGRP蛋白表达降低,中脘组、内关-中脘组Ghrelin蛋白表达上升、CGRP蛋白表达降低(
P
<0.01或
P
<0.05)。与内关-中脘组比较,中脘组、内关组Ghrelin蛋白表达显著降低,CGRP蛋白表达显著升高(
P
<0.05或
P
<0.01)。代谢组学结果显示,与正常组比较,模型组大鼠胃窦组织中代谢物二磷酸腺苷核糖、一磷酸腺苷、二磷酸腺苷等含量下降;与模型组比较,内关组二磷酸腺苷核糖、二磷酸腺苷、胞磷胆碱含量上升,中脘组烟碱腺嘌呤二核苷酸、胞苷二磷酸乙醇胺、胞磷胆碱等含量升高,内关-中脘组二磷酸腺苷核糖、胞苷二磷酸乙醇胺、胞磷胆碱等含量上升(
P
<0.05或
P
<0.01)。与模型组比较,内关-中脘组大鼠差异性代谢产物的主要代谢通路分别为甘油磷脂代谢、牛磺酸和次牛磺酸代谢、嘌呤代谢、淀粉和蔗糖代谢。
结论
2
电针“内关”“中脘”穴组能有效调节FD大鼠胃肠动力,改善FD症状,作用优于单用“内关”“中脘”穴,其作用机制可能与相关差异代谢物影响能量代谢、糖代谢、核苷酸代谢,从而调节胃肠动力激素有关。
Objective
2
To investigate the possible mechanism of electroacupuncture at "Neiguan" (PC6) and "Zhongwan" (RN12) in the treatment of functional dyspepsia (FD).
Methods
2
SPF male SD rats were randomly divided into model group, Neiguan (PC6) group, Zhongwan (RN12) group, and Neiguan-Zhongwan (PC6-RN12) group, with 8 rats in each group. Except for the normal group, the other groups of rats were cogavaged with 0.1% sucrose iodoacetamide solution combined with small platform standing training to establish FD rat models. After successful modeling, the rats in the normal group and model group were tied up for 30 min/d for 7 days; the Neiguan group, Zhongwan group, and Neiguan-Zhongwan group were treated with electroacupuncture intervention at "Neiguan" (PC6), "Zhongwan" (RN12), and "Neiguan" - "Zhongwan" (PC6-RN12) acupoints, respectively, using continuous wave, frequency of 2 Hz, current intensity of 1 mA, 30 min/d, and continuous intervention for 7 days. The general condition of rats in each group was observed. After treatment, the body weight and food intake of rats were measured, and the gastric emptying rate was calculated; HE staining was performed on the gastric antrum tissue of rats to observe the histopathologic changes; the expressions of calcitonin gene-related peptide (CGRP) and Ghrelin protein in gastric antrum were detected by Western Blot; the metabolites in gastric antrum tissues were detected by liquid chromatography-mass spectrometry (LC-MS), and differential metabolites were screened by correlation analysis, then Metabo Analyst 5.0 and KEGG databases were used for metabolic pathway analysis.
Results
2
Under light microscope, the gastric antrum structure was complete and the glands were abundant. No obvious inflammation and edema were found in gastric mucosa. Compared with the normal group, the body weight, food intake, and gastric emptie rate of rats in model group decreased, the expression of Ghrelin protein decreased and the expression of CGRP protein increased in gastric antrum tissue (
P
<0.01). Compared with model group, the body weight, food intake, and gastric emptance rate of rats in Neiguan group, Zhongwan group and Neiguan-Zhongwan group all increased, CGRP protein expression decreased in Neiguan group, and Ghrelin protein expression increased and CGRP protein expression decreased in Zhongwan group and Neiguan-Zhongwan group (
P
<0.01 or
P
<0.05). Compared with Neiguan-Zhongwan group, Ghrelin protein expression decreased and CGRP protein expression increased in Neiguan group and Zhongwan group (
P
<0.05 or
P
<0.01). Metabolomics results showed that compared with normal group, the content of metabolites adenosine diphosphate ribose, adenosine monophosphate and adenosine diphosphate in gastric antrum tissue of model group decreased; compared with model group, the contents of adenosine diphosphate, adenosine diphosphate and citicoline in Neiguan group increased, the contents of nicotine adenine dinucleotide, cytidine diphosphate ethanolamine and citicoline in Zhongwan group increased, and the contents of adenosine diphosphate, cytidine diphosphate and citicoline in Neiguan-Zhongwan group increased (
P
<0.05 or
P
<0.01). C
ompared with model group, the main metabolic pathways of different metabolites in PC6-RN12 group were glycerophospholipid metabolism, taurine and hypotaurine metabolism, purine metabolism, starch and sucrose metabolism.
Conclusion
2
Electroacupuncture at “Neiguan” and “Zhongwan” acupoints can effectively regulate gastrointestinal motility and improve FD symptoms in FD rats, and the effect is better than that of "Neiguan" or "Zhongwan" acupoints alone. The mechanism may be related to the influence of related metabolites on energy metabolism, glucose metabolism and nucleotide metabolism, thereby regulating gastrointestinal motility hormones.
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