基于NGF/TrKA/TRPV1通路探讨电针改善功能性消化不良大鼠胃高敏感性

doi:10.3969/j.issn.1006-5725.2023.22.012

摘要/Abstract

摘要：

目的探讨电针对功能性消化不良（FD）大鼠胃高敏感性的作用效果及机制。方法将30只大鼠随机平均分为空白组、模型组、电针组、抑制剂组、电针+抑制剂组。空白组不予以特殊处理，其余各组采用多因素刺激法进行造模。造模完成后，模型组常规饲养，电针组和电针+抑制剂组予以每天电针足三里，抑制剂和电针+抑制剂组予以每天腹腔注射神经生长因子（NGF）抑制剂。采用胃内球囊置入检测胃高敏感性和胃顺应性，采用甲胺蓝染色观察胃肥大细胞活化程度，采用免疫组化法检测胃瞬时受体电位香草酸亚型1（TRPV1）定位表达，采用免疫印迹检测胃NGF/原肌球蛋白受体激酶（TrkA）/TRPV1蛋白表达，采用酶联免疫吸附检测胃降钙素基因相关肽（CGRP）含量。结果相比于空白组，模型组大鼠体重增长缓慢，胃敏感性增高、顺应性降低，胃肥大细胞数和脱颗粒率显著增加，NGF、TrKA、TRPV1的蛋白表达和CGRP含量显著升高；与模型组比较，电针组、抑制剂组和电针+抑制剂组大鼠体重增加，胃高敏感性和顺应性显著改善，肥大细胞数目和活化程度显著降低，胃NGF/TrKA/TRPV1蛋白表达和CGRP含量显著降低；且相比于电针组和抑制剂组，电针+抑制剂组胃NGF/TrKA/TRPV1蛋白表达和CGRP含量进一步降低，差异有统计学意义。结论电针可能是通过调控肥大细胞介导的NGF/TrKA/TRPV1通路改善FD大鼠胃高敏感性。

关键词: 功能性消化不良, 胃高敏感性, 肥大细胞, 神经生长因子/原肌球蛋白受体激酶, 瞬时受体电位香草酸亚型1

Abstract:

Objective To explore the effect and mechanism of electroacupuncture （EA） on gastric hypersensitivity in rats with functional dyspepsia （FD）. Methods Rats were randomly divided into blank group， model group， EA group， inhibitor group and EA+inhibitor group. The blank group was not subjected to special treatment， and the model group， EA group， inhibitor group， and EA+inhibitor group were modeled using a multi factor stimulation method. After the modeling， the model group was given routine feeding， the EA group and the EA+inhibitor group were treated with daily electroacupuncture at Zusanli， and the inhibitor group and the EA+ inhibitor group were treated with daily intraperitoneal injection of anti-nerve growth factor（NGF）. Gastric hypersensitivity and gastric compliance were detected by intragastric balloon implantation， the degree of mast cell activation were observed by methylamine blue staining， the localization expression of gastric transient receptor potential vanillin subtype 1（TRPV1） was detected by immunohistochemistry， the expression of gastric NGF/tropomyosin receptor kinase （TrkA）/TRPV1 protein was detected by Western blot， and the content of gastric calcitonin gene-related peptide （CGRP） was detected by enzyme-linked immunosorbent assay. Results Compared with the blank group， the weight of the model group rats increased slowly， gastric sensitivity increased， compliance decreased， the number of gastric mast cell and degranulation rate increased significantly， and the protein expression and CGRP content of NGF， TrKA， TRPV1 increased significantly. Compared with the model group， the rats in the EA group， the inhibitor group and the EA+inhibitor group gained weight， improved gastric hypersensitivity and compliance significantly， reduced the number and activation of mast cell significantly， and reduced gastric NGF/TrKA/TRPV1 protein expression and CGRP content significantly. Compared with the EA group and the inhibitor group， the expression of NGF/TrKA/TRPV1 protein and CGRP content in the stomach of the EA+inhibitor group were further reduced， and the difference was statistically significant. Conclusion EA may relieve the gastric hypersensitivity of FD rats by regulating the NGF/TrKA/TRPV1 pathway activated by mast cells.

Key words: functional dyspepsia, gastric hypersensitivity, mast cells, nerve growth factor/tropomyosin receptor kinase, transient receptor potential vanilloid type 1

中图分类号:

R245.3

金舒文,刘伟,刘嘉宝,范建超,韩永丽,周丽,徐派的,张红星. 基于NGF/TrKA/TRPV1通路探讨电针改善功能性消化不良大鼠胃高敏感性[J]. 实用医学杂志, 2023, 39(22): 2928-2933.

Shuwen JIN,Wei LIU,Jiabao LIU,Jianchao FAN,Yongli HAN,Li ZHOU,Paidi XU,Hongxing. ZHANG. Mechanism of electroacupuncture for gastric hypersensitivity in functional dyspepsia rats rats through NGF/TrKA/TRPV1 pathway[J]. The Journal of Practical Medicine, 2023, 39(22): 2928-2933.

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参考文献 23

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组别	例数	基线期	造模后	干预后
空白组	6	16.36 ± 0.69	263.65 ± 7.50	381.12 ± 21.00
模型组	6	17.20 ± 0.91	179.07 ± 13.62^*	260.20 ± 8.97^*
电针组	6	16.83 ± 0.74	169.05 ± 11.00^*	304.00 ± 13.14^*#
抑制剂组	6	17.17 ± 0.52	172.58 ± 12.20^*	291.58 ± 12.21^*#
电针+抑制剂组	6	16.65 ± 0.68	171.28 ± 9.82^*	306.38 ± 14.33^*#
F值		1.20	68.27	43.11
P值		0.334	< 0.001	< 0.001

组别	例数	20 mmHg	40 mmHg	60 mmHg	80 mmHg
空白组	6	0.16 ± 0.01	0.20 ± 0.02	0.25 ± 0.02	0.27 ± 0.01
模型组	6	0.14 ± 0.02	0.17 ± 0.01^*	0.19 ± 0.01^*	0.18 ± 0.01^*
电针组	6	0.15 ± 0.01	0.19 ± 0.01^#	0.23 ± 0.02^#	0.22 ± 0.01^#
抑制剂组	6	0.15 ± 0.02	0.19 ± 0.01^#	0.22 ± 0.01^#	0.21 ± 0.02^#
电针+抑制剂组	6	0.15 ± 0.01	0.19 ± 0.01^#	0.23 ± 0.01^#	0.23 ± 0.01^#
F值		1.294	6.209	14.08	32.84
P值		0.2993	< 0.01	< 0.001	< 0.001

组别	例数	20 mmHg	40 mmHg	60 mmHg	80 mmHg
空白组	6	0.33 ± 0.47	1.33 ± 0.47	2.00 ± 0.82	2.67 ± 0.47
模型组	6	0.67 ± 0.47	2.83 ± 0.69^*	3.50 ± 0.50^*	3.83 ± 0.37^*
电针组	6	0.5 ± 0.5	2.33 ± 0.47	2.50 ± 0.50^#	3.16 ± 0.37^#
抑制剂组	6	0.67 ± 0.47	2.67 ± 0.75	2.67 ± 0.47^#	3.16 ± 0.37^#
电针+抑制剂组	6	0.83 ± 0.37	2.17 ± 0.69	2.33 ± 0.47^#	3.00 ± 0.58^#
F值		0.86	4.383	12.92	4.643
P值		0.50	< 0.01	< 0.001	< 0.01

组别	例数	肥大细胞记数（个）	肥大细胞脱颗粒率（%）
空白组	6	4.83 ± 2.67	16.67 ± 13.05
模型组	6	16.50 ± 3.99^*	67.64 ± 15.11^*
电针组	6	9.00 ± 4.00^#	31.84 ± 16.21^#
抑制剂组	6	10.50 ± 4.37^#	38.73 ± 11.00^#
电针+抑制剂组	6	7.83 ± 4.37^#	25.97 ± 14.50^#
F值		6.094	9.449
P值		< 0.01	< 0.001

组别	例数	TRPV1平均光密度值
空白组	6	0.013 ± 0.001
模型组	6	0.063 ± 0.008^*
电针组	6	0.037 ± 0.005^#△
抑制剂组	6	0.039 ± 0.005^#△
电针+抑制剂组	6	0.025 ± 0.004^#
F值		64.71
P值		< 0.001