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[用于研究针刺效应的皮肤切片模型]

[Skin slice used as a model for investigating acupuncture effects].

作者信息

Wang Li-Na, Schwarz Wolfgang, Gu Quan-Bao, Ding Guang-Hong

机构信息

Shanghai Research Center of Acupuncture and Meridians, Department of Mechanics and Engineering Science, Fudan University, Shanghai 200433, China.

出版信息

Zhen Ci Yan Jiu. 2009 Oct;34(5):291-6.

Abstract

OBJECTIVE

To establish an acupoint-connective tissue model for studying the mechanism of acupuncture by using in vitro patch clamp technique in the rat skin slices.

METHODS

The local connective tissue under the corium of "Housanli" (ST 36) area from SD rat was acutely and bluntly separated and fixed in a chamber filled with artificial incubation solution. Mast cells in the prepared connective tissue slice were labeled by toluidine blue (TB) or neutral red (NR). The whole-cell current of mast cells responding to pressure stimulation applied through a patch pipette was recorded in rat slices derived from acupoint ST 36 area by using in vitro patch-clamp technique.

RESULTS

  1. After staining with TB and NR, the labeled mast cells were found to distribute in the extracellular matrix of the connective tissue samples, and their degranulation phenomenon could be seen clearly. 2) The whole-cell current of mast cells in response to mechanical stress stimulation was successfully recorded in the connective tissue slices of the rat acupoint ST 36 area. The cellular membrane currents increased evidently when pressure gradients of -30, -60 or -90 cmH2O were applied to the recorded mast cells.

CONCLUSION

The connective tissue slice from the rat ST 36 area may be used as a model for investigating the peripheral mechanism of acupuncture by combining the microtechniques and electrophysiological techniques. The results obtained in this model prove for the first time by electrophysiology that the mast cells in the connective tissue are probably involved in the transduction process of the mechanical signal from acupuncture stimulation. This new model provides a base for investigating the characters of the cells, collagen fibers, proteoglycans, etc. and their interactions in the acupoint connective tissue in the future.

摘要

目的

利用体外膜片钳技术,在大鼠皮肤切片上建立穴位-结缔组织模型,以研究针刺机制。

方法

将SD大鼠“足三里”(ST 36)区域真皮层下的局部结缔组织进行急性钝性分离,并固定于充满人工孵育液的槽中。用甲苯胺蓝(TB)或中性红(NR)对制备的结缔组织切片中的肥大细胞进行标记。采用体外膜片钳技术,在源自穴位ST 36区域的大鼠切片中记录肥大细胞对通过膜片移液管施加的压力刺激的全细胞电流。

结果

1)经TB和NR染色后,发现标记的肥大细胞分布于结缔组织样本的细胞外基质中,且其脱颗粒现象清晰可见。2)在大鼠穴位ST 36区域的结缔组织切片中成功记录到肥大细胞对机械应力刺激的全细胞电流。当对记录的肥大细胞施加-30、-60或-90 cmH2O的压力梯度时,细胞膜电流明显增加。

结论

大鼠ST 36区域的结缔组织切片可作为结合显微技术和电生理技术研究针刺外周机制的模型。该模型所获结果首次通过电生理学证明,结缔组织中的肥大细胞可能参与针刺刺激机械信号的转导过程。这一新模型为今后研究穴位结缔组织中细胞、胶原纤维、蛋白聚糖等的特性及其相互作用提供了基础。

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