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基于核磁共振波谱法探讨艾灸对胃热证和胃寒证大鼠损伤胃黏膜组织影响的内源性代谢产物

[Endogenous metabolites involving effects of moxibustion on injured gastric mucosal tissue in rats with stomach heat or cold syndrome based on nuclear magnetic resonance spectroscopy].

作者信息

Yang Jin-Lan, Cai Run-Tian, Yu Yun-Jin, Wei Gu-Hang, Xie Yu-Feng, Feng Jun, Yang Zong-Bao

机构信息

Shenzhen Hospital of Guangzhou University of Chinese Medicine (Futian), Shenzhen 518048, Guangdong Province, China.

The First Clinical Medical College, Guangzhou University of Chinese Medicine, Guangzhou 510405.

出版信息

Zhen Ci Yan Jiu. 2020 Jun 25;45(6):446-53. doi: 10.13702/j.1000-0607.190480.

Abstract

OBJECTIVE

To investigate the profile of metabolites of gastric mucosa involving the effectiveness of moxibustion in the treatment of syndromes of stomach heat (SH) and stomach cold (SC) by H-nuclear magnetic resonance (H-NMR) spectroscopy in rats, so as to reveal its mechanisms underlying improvement of gastric disorders.

METHODS

Male SD rats were randomly divided into control, SH-model, SC-model, SH-moxibustion and SC-moxibustion groups (=6 rats/group). The SH-model and SC-model were established by gavage of pepper liquid plus ethanol, and ice water plus NaOH, respectively. Moxibustion was applied to bilateral "Zusanli" (ST36) and "Liangmen"(ST21) for 20 min, once daily for 7days. Histopathological changes of the gastric tissue were observed by H.E. staining. Differential metabolites in the gastric mucosal tissue were detected and the relevant metabolic pathways analyzed by using H-NMR, pattern recognition method,and online MetPA (http: //www.metaboanalyst.ca).

RESULTS

Compared with the control group, the body mass was decreased significantly from the 4 to 14 day after modeling (<0.05,<0.01). After the treatment, the body mass was obviously increased from the 10 day on in both SH-EA and SC-EA groups relevant to the SH and SC model group, respectively (<0.05,<0.01). H.E. staining showed severe damage of the columnar epithelial structure of the gastric mucosa and inflammatory cell infiltration in the SH group, and inflammatory cell infiltration in the SC model group, which were relatively milder in both moxibustion groups. H-NMR analysis displayed a total of 16 potential biomarkers in the injured gastric mucosa of SH syndrome and 14 biomarkers for the SC syndrome after mode-ling, and 13 metabolites related to SH moxibustion and 8 metabolites related to SC moxibustion after moxibustion interventions, respectively. After moxibustion, among the 13 differential metabolites of the SH syndrome, the effectively up-regulated candidates were isoleucine, creatinine, choline and lactate (<0.05), and the down-regulated ones were choline phosphate, glycine, alanine, urine pyrimidine, tyrosine, phenylalanine, hypoxanthine, adenosine and nicotinamide (<0.05). Among the 8 metabolites related to the SC syndrome, creatinine, ethanolamine, choline, adenosine and nicotinamide were markedly increased (<0.05), and glycine, creatine phosphate and tyrosine remarkably decreased in their levels after moxibustion (<0.05). MetPA showed that moxibustion could regulate 10 metabolic pathways for SH syndrome and 7 metabolic pathways for SC syndrome. Metabolites and metabolic pathways are mainly involved in functions of amino acid metabolism, energy metabolism and inflammatory response.

CONCLUSION

The idea of "moxibustion could be used for heat syndrome" has metabolic substance basis, and its efficacy in repairing the injured gastric mucosa involves regulation of amino acid metabolism, energy balance and inflammation response, and moxibustion for SH and SC syndromes has both generality and specificity in regulating metabolic activities.

摘要

目的

采用氢核磁共振(H-NMR)波谱技术研究胃黏膜代谢产物谱,探讨艾灸对大鼠胃热证和胃寒证的治疗作用机制,揭示艾灸改善胃功能紊乱的潜在机制。

方法

将雄性SD大鼠随机分为对照组、胃热模型组、胃寒模型组、胃热艾灸组和胃寒艾灸组(每组6只)。胃热模型组和胃寒模型组分别通过灌胃辣椒液加乙醇、冰水加氢氧化钠建立。艾灸双侧“足三里”(ST36)和“梁门”(ST21)20分钟,每日1次,共7天。采用苏木精-伊红(H.E.)染色观察胃组织病理变化。利用H-NMR、模式识别方法及在线MetPA(http://www.metaboanalyst.ca)检测胃黏膜组织中的差异代谢产物并分析相关代谢途径。

结果

与对照组相比,造模后4至14天体重显著下降(P<0.05,P<0.01)。治疗后,胃热艾灸组和胃寒艾灸组分别从第10天起体重较胃热模型组和胃寒模型组明显增加(P<0.05,P<0.01)。H.E.染色显示,胃热组胃黏膜柱状上皮结构严重损伤,有炎性细胞浸润;胃寒模型组有炎性细胞浸润,艾灸组上述改变相对较轻。H-NMR分析显示,造模后胃热证损伤胃黏膜中有16种潜在生物标志物,胃寒证有14种生物标志物;艾灸干预后,胃热艾灸组有13种代谢产物,胃寒艾灸组有8种代谢产物。艾灸后,胃热证的13种差异代谢产物中,异亮氨酸、肌酐、胆碱和乳酸有效上调(P<0.05),磷酸胆碱、甘氨酸、丙氨酸、尿嘧啶、酪氨酸、苯丙氨酸、次黄嘌呤、腺苷和烟酰胺下调(P<0.05)。胃寒证的8种代谢产物中,肌酐、乙醇胺、胆碱、腺苷和烟酰胺显著增加(P<0.05),甘氨酸、磷酸肌酸和酪氨酸水平明显降低(P<0.05)。MetPA显示,艾灸可调节胃热证10条代谢途径,胃寒证7条代谢途径。代谢产物和代谢途径主要涉及氨基酸代谢、能量代谢及炎症反应功能。

结论

“热证可灸”有代谢物质基础,艾灸修复损伤胃黏膜的作用机制涉及调节氨基酸代谢、能量平衡及炎症反应,艾灸对胃热证和胃寒证在调节代谢活动方面既有共性又有特性。

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