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J Acupunct Res > Volume 32(3); 2015 > Article
Lee, Rho, and Kim: Effects of Acupuncture at SP6 on Reflux Esophagitis in Rats※
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Abstract

Objectives:

The purpose of this study was to evaluate whether acupuncture at SP6 attenuates esophageal inflammation on refluxed-induced esophagitis.

Methods:

Acupuncture at SP6 was stimulated by acupuncture torsion technique for 30 seconds four times every hour after an operation induced reflux esophagitis(RE), and its effects were assessed in comparison with RE rats without acupuncture, and normal rats.

Results:

SP6 acupuncture stimulation markedly ameliorated mucosal damage in the histological evaluation. Reflux-induced esophagitis rats exhibited the down-regulation of antioxidant-related protein expression levels such as heme oxygenase-1(HO-1) in the esophagitis; however, the associated levels with SP6 acupuncture stimulation were significantly higher than those in RE rats without acupuncture stimulation. Moreover, SP6 acupuncture stimulation significantly reduced the expression of inflammatory proteins through mitogen-activated protein kinase(MAPK)- related signaling pathways. The increased protein expressions of inflammatory mediators, cyclooxygenase-2(COX-2) and inducible nitric oxide synthase(iNOS), by nuclear factor-kappa B(NF-kB) activation were significantly suppressed through SP6 acupuncture stimulation.

Conclusions:

Our findings support the therapeutic evidence for SP6 acupuncture stimulation alleviating the development of esophagitis via regulating inflammation through the activation of the antioxidant pathway.

Fig. 1
Gross evaluation of the esophageal mucosal damage
A: Representative microphotographs of the esophagus. Esophageal lesion observed in RE rats was ameliorated by the acupuncture stimulation at SP6.
B: Gross mucosal injury ratio at the end of experiment. The gross mucosal injury was increased in RE rats compared with normal rats, but SP6 acupuncture stimulation led to a significant decrease.
N: normal rats.
Veh: RE rats without acupuncture stimulation.
SP6: RE rats and SP6 acupuncture stimulation. Values are the means ± SEM.
*: p<0.01, **: p<0.001.
v. RE rats without acupuncture stimulation values. n=6 in each group.
acupunct-32-3-83f1.gif
Fig. 2
Histophasological changes in esophagus tissue
RE control group examined the removed mucosa, edematous submucosa by inflammation(yellow arrow), and infiltration of inflammatory cells in muscularis externa(red arrow). Whereas, SP6 group, still occurred submucosa edema(black arrow), protected the esophageal mucosa damage and had little the infiltration of inflammatory cells in ME.
A·B: esophagus tissue stained with H&E on normal rat.
C·D: esophagus tissue stained with H&E on reflux esophagitis rats.
E·F: esophagus tissue stained with H&E on reflux esophagitis rats treated with SP6.
A·C and D are captured images with isolution software with 200 magnification on Olympus microscope.
B·D and F are images captured with isolution software with 400 magnification.
acupunct-32-3-83f2.gif
Fig. 3
Effect of gastric contents pH and gastric volume
Gastric pH and volume analyses were performed as described in Materials and Methods.
N: normal rats.
Veh: RE rats without acupuncture stimulation.
SP6: RE rats with SP6 acupuncture stimulation.
Values are the means ± SEM.
*: p<0.01. **: p<0.001.
vs. normal rat values.
n=6 in each group.
acupunct-32-3-83f3.gif
Fig. 4
Esophageal HO-1 protein expression
Immunoblotting analyses were performed as described in materials and methods.
N: normal rats.
Veh: RE rats without acupuncture stimulation.
SP6: RE rats with SP6 acupuncture stimulation.
Values are the means ± SEM.
*: p<0.01.
vs. RE rat without acupuncture stimulation values.
n=6 in each group.
acupunct-32-3-83f4.gif
Fig. 5
Esophageal p-p38(A) and p-ERK(B) protein expressions.
p-p38: phosphor-p38.
p-ERK: phosphorylated extracellular signal-regulated kinase.
Immunoblotting analyses were performed as described in materials and methods.
N: normal rats.
Veh: RE rats without acupuncture stimulation.
SP6: RE rats with SP6 acupuncture stimulation.
Values are the means ± SEM.
*: p<0.05. **: p<0.01. ***: p<0.001.
vs. RE rat without acupuncture stimulation values.
n=6 in each group.
acupunct-32-3-83f5.gif
Fig. 6
Esophageal NF-kBp65(A), COX-2(B), and iNOS(C) protein expressions.
Immunoblotting analyses were performed as described in materials and methods.
N: normal rats. Veh: RE rats without acupuncture stimulation. SP6: RE rats with SP6 acupuncture stimulation.
Values are the means ± SEM.
*: p<0.01. **: p<0.001.
vs. RE rat without acupuncture stimulation values.
n=6 in each group.
acupunct-32-3-83f6.gif
Table 1
Effect of SP6 Acupuncture Treatment on Esophageal Histomorphometry
Group Protecting percentage of mucosa(%) Thickness o of mucosa(μm)
N 99.6 ± 0.5*** 259.2 ± 31.2***
RE Veh 25.9 ± 7.1 22.2 ± 19.0
SP6 53.0 ± 14.2* 151.5 ± 38.2**

N: normal rats.

Veh: RE rats without acupuncture stimulation.

SP6: RE rats with SP6 acupuncture stimulation.

Values are the means ± SEM.

* p<0.05,

** p<0.01,

*** p<0.001.

vs. RE rat without acupuncture stimulation values, n=6 in each group.

VI.
VI.

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