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1、 NEURAL REGENERATION RESEARCH Volume 7, Issue 30, October 2012 doi:10.3969/j. issn. 1673-5374.2012.30.007 http:/www.crter.org/nrr-2012-qkquanwen.html Peng L, Liu M, Chang XR, Yang Z, Yi SX, YanJ, Peng Y. Effects of pre-moxibustion at Zusanli (ST36) on heat shock protein 70 expression in rats with ga

2、stric mucosal lesions after neurotomy. Neural Regen Res. 2012;7(30):2370-2376. Effects of pre-moxibustion at Zusanli (ST36) on heat shock protein 70 expression in rats with gastric mucosal lesions after neurotomy* Liang Peng, Mi Liu, Xiaorong Chang, Zhou Yang, Shouxiang Yi, Jie Yan, Yan Peng College

3、 of Acupuncture and Tuina, Hunan University of Chinese Medicine, Changsha 410007, Hunan Province, China Abstract Studies have shown that pre-moxibustion protects the gastric mucosa by up-regulating the expres- sion of heat shock protein 70. However, the signaling pathway underlying this effect remai

4、ns unclear. Rats were intragastrically administered absolute alcohol, causing obvious lesion of the gastric mu- cosa. Following pre-moxibustion at Zusanli (ST36) for 8 days, the ulcer index decreased to different degrees. The results of an enzyme linked immunosorbent assay and western blotting showe

5、d sig- nificant upregulation of heat shock protein 70 expression in the gastric mucosa and serum. None out of transection of the spinal cord, damage to the nucleus of the solitary tract, neurotomy of the vagal nerve and neurotomy of the common peroneal nerve affected the decrease in ulcer index or t

6、he increase in heat shock protein 70 expression in serum after pre-moxibustion at Zusanli, and heat shock protein 70 expression was obviously decreased in the gastric mucosa. These findings suggest that pre-moxibustion at Zusanli can protect the gastric mucosa against lesioning, and that the mechani

7、sm underlying this effect involves its induction of heat shock protein 70 expression. Neural pathways participate in the regulatory effects of moxibustion on heat shock protein 70 ex- pression in the gastric mucosa. Liang Peng, Ph D., Lecturer, College of Acupuncture and Tuina, Hunan University of C

8、hinese Medicine, Changsha 410007, Hunan Province, China Corresponding author: Xiaorong Chang, Professor, Doctoral supervisor, College of Acupuncture and Tuina, Hunan University of Chinese Medicine, Changsha 410007, Hunan Province, China xrchang 1956 Received: 2012-03-15 Accepted: 2012-06-24 (N20120

9、117001/YJ) Key Words pre-moxibustion; transection of neural pathway; Zusan/i(ST36) gastric mucosal lesion; heat shock proteins-70; ulcer index; traditional Chinese medicine; neural regeneration Research Highlights (1) Moxibustion stimulation transformed into bioinformation related to outcomes activa

10、ted the protective potential of the body through various pathways, and protected the gastric mucosa against lesion. (2) This study explored the correlation between endogenous protective information and neural pathways by studying the influence of the afferent pathways of damaged nerves on the protec

11、tive effects of moxibustion. INTRODUCTION Chemical and physical factors, microorganism infection, bacterial toxins, and oral drugs can cause acute inflammatory reaction of the gastric mucosa, lead to gastric mucosal lesions. After formation of gastric mucosal lesions, cell proliferation occurs at mu

12、ltiple areas around the bottom and cervical parts of the gastric gland13. Among members of the heat shock protein family, heat shock protein 70 is involved in the stress response, and is important for protection of the gastric mucosa46. Nisitani ef a/m first showed 2370 Peng L, et a!. /NeuralRegener

13、ation Research. 2012;7(30):2370-2376. that cultured gastric mucosal cells can resist excoriation and acute necrosis, caused by ethanol of lethal concentration, after heat shock treatment, which induces heat shock protein expression. Heat shock protein 70 is strongly expressed in post-transplanted al

14、vine mucosal epithelium of the small intestine and inflammatory cells; therefore, heat shock protein 70 is considered to play an important role in the regeneration of the mucous membrane8. The inducement of heat shock protein in rat stomach occurs in proportion to the protection of the gastric mucos

15、a9. Tsukimi ef a/10 reported that heat shock protein 70 expression in the ulcer margin was gradually increased during ulcer healing. Overexpression of heat shock protein 70 in the stomach seems to protect against gastric ulcers through its cytoprotective effects on the gastric mucosa by increasing m

16、ucosal blood flow11. Accumulation of heat shock protein 70 can accelerate the restoration of the gastric mucosa through regulating apoptosis and rebuilding cytoarchitecture12. Therefore, heat shock protein 70 is considered to play an important role in the recovery from gastric mucosal lesions1316. P

17、re-moxibustion is named uNi-jiun in Chinese. The Chinese term means using the method of pre-moxibustion to guard against disease. It is a traditional Chinese medicine treatment that aims to excite the Q/in channels, to promote immunity and irritability, and to prevent sickness and alleviate the inju

18、ry caused by illness using moxibustion when disease has not occurred or the illness is minor. Modern traditional Chinese medicine experts and practitioners have studied the effects of and mechanisms underlying pre-moxibustion warming and invigorating of the spleen and stomach from various angles. Pr

19、e-moxibustion to prevent gastric mucosal lesions is an effective naturopathy, which has been used from ancient to modern times. The modern theory of stress considers that pre-moxibustion can arouse body potentials, and that it can also arouse and regulate intrinsic resistance against diseases. In th

20、ese processes, the irritated responsiveness can mobilize an intrinsic prevention mechanism, part of the defense reflex17. The study of the effects of pre-moxibustion on gastric mucosal lesions and the underlying mechanisms has become a research hotspot in the field of research into the mechanisms by

21、 which moxibustion prevents sickness1820. The nervus peroneus communis, spinal cord, nucleus of the solitary tract and vagus nerve are all involved in transducing signals from the splanchnic nerves2123. In this study, neurotomy of these parts of the rats central and peripheral nervous systems was pe

22、rformed; then, we carried out pre-moxibustion at acupoint Zusan/i(ST36). After acute gastric mucosal lesion models were made, endogenous heat shock protein 70 in the gastric mucosa and serum was examined. Using this method, the relationship between the effect of pre-moxibustion on gastric mucosal le

23、sions and neural regulation was explored. RESULTS Quantitative analysis of experimental animals A total of 70 Sprague-Dawley rats were randomly divided into seven groups: control, model, moxibustion, moxibustion + transection of spinal cord, moxibustion + damaged nucleus of the solitary tract, moxib

24、ustion + vagotomy and moxibustion + neurotomy of the common peroneal nerve. Neurotomy of the spinal cord, nucleus of solitary tract, vagus, and common peroneal nerve were performed in rats from the last four groups before pre-moxibustion at acupoint Zusan/iuas carried out for 8 days. Acute gastric m

25、ucosal lesion models were made in all groups except the control group. All 70 rats were involved in the final analysis. Changes in gastric mucosal lesions following transection of various neural pathways and moxibustion pretreatment As shown in Table 1, considerable damage appeared in the gastric mu

26、cosa of animals in the model group. Damage in the gastric mucosa of animals in the moxibustion, moxibustion + transection of spinal cord, moxibustion + damaged nucleus of solitary tract, moxibustion + vagotomy and moxibustion + neurotomy of common peroneal nerve groups was decreased compared with th

27、at in the model group. The amounts of damage in the gastric mucosa in the moxibustion + transection of spinal cord, moxibustion + damaged nucleus of the solitary tract, moxibustion + vagotomy, and moxibustion + neurotomy of the common peroneal nerve groups were greater than that in the moxibustion g

28、roup. Heat shock protein 70 expression in the serum and gastric mucosa following transection of various neural pathways There was no difference in the level of expression of heat shock protein 70 in the gastric mucosa between the model and control groups. The level of expression of heat shock protei

29、n 70 in the serum and gastric mucosa of animals in the moxibustion, moxibustion + transection of the spinal cord, moxibustion + damaged nucleus of the solitary tract, moxibustion + vagotomy and moxibustion + neurotomy of the common peroneal nerve groups was significantly greater than the levels in t

30、he model group. The level of heat shock protein 2371 Peng L, et a!. /NeuralRegeneration Research. 2012;7(30):2370-2376. 70 expression in the serum of operation groups (mox- ibustion + transection of the spinal cord, moxibustion + damaged nucleus of the solitary tract, moxibustion + vagotomy, and mox

31、ibustion + neurotomy of the common peroneal nerve group) was not significantly different from that in the moxibustion group, but the level of heat shock protein 70 expression in the gastric mucosa of operation groups was lower than that in the moxibustion group (Table 1, Figure 1). Table 1 Ulcer ind

32、ex and heat shock protein 70 (HSP70) expression i 门 serum and the gastric mucosa Ulcer index Group (mm) HSP70 in serum (MQ/L) HSP70 in the gastric mucosa (relative value of absorbance/(3-actin) Control 0 176.2216. 95 0.520.05 Model 66.915.1 177.6819.14 0.540.03 Moxibustion 16.18.1b 222.3449.40a 0.74

33、0.06a Moxibustion + 46.614.0bc tr3 门 section of spinal cord 201,4025.78a 0.620.06bc Moxibustion + 51.217.2ac damaged 门 ucleus of solitary tract 217.8230.64a 0.640.07bd Moxibustion + 42.821.5bc vagotomy 236.4532.96b 0.580.04ac Moxibustion + 32.914.4bc neurotomy of common peroneal nerve 219.3444.25a 0

34、.630.04bc The gastric mucosal impairment site length was calculated as the ulcer index and expressed in millimeters. Ulcer index was doubled if the width of injury was more than 2 mm. The data are analyzed using one-way analysis of variance, and are expressed as mean SD. For assessment of the ulcer

35、index and HSP70 level in serum, we used 10 rats from each group; for assessment of HSP70 expression in the gastric mucosa, we used six rats from each group. aP 0.05, bP 0.01, vs. model group; CP 0.01, dP 0.05, vs. moxibustion group. DISCUSSION A previous study showed that moxibustion at Zusan/i and

36、Liangmen (ST2 has a protective effect on the gastromucosa in stressed rats with gastric ulcers, and that this effect is strongly associated with its actions in promoting the synthesis of tumor growth factor-a and proliferation of gastromucosal cells, suppressing apoptosis of gastric mucosa cells and

37、 upregulating heat shock protein 70 mRNA expression1-241. Moxibustion at Zusanli and Hangmen can protect the gastric mucosa through inducing high expression of heat shock protein 70 protein and mRNA, with relative specificity of the acupoints20. Figure 1 Heat shock protein 70 (HSP70) expression in t

38、he gastric mucosa (Western blot: HSP70, 70 kDa; p-actin, 43 kDa). Moxibustion can protect the gastric mucosa from oxidative injury by inducing heat shock protein 70 expression with relative specificity of acupoints25. Moxibustion may up-regulate heat shock protein 70 expression in the gastric mucosa

39、, and then act on targets associated with the mitochondrial signal transduction pathway to inhibit gastric mucosal cell apoptosis and protect the gastric mucosa. This protective effect is acupoint specific to some extent26. While heat shock protein 70 expression in the gastric mucosa was up-regulate

40、d by treatment with pre-moxibustion, cell apoptosis was suppressed in rats with acute gastric mucosal injuries. However, after heat shock protein 70 expression was blocked by quercetin, the effect of pre-moxibustion on apoptosis was canceled. This study confirmed that pre-moxibustion has a protectiv

41、e effect on the gastric mucosa, and that the effect is correlated with up-regulation of heat shock protein 70 expression18. Furthermore, moxibustion at Zusan/iar6 Hangmen has been shown to increase heat shock protein 70 expression in organs and heat shock protein 70 mRNA levels in the rat gastric mu

42、cosa, which may be one of the mechanisms by which it promotes health27. In summary, some studies have suggested that the mechanism underlying the effect of pre-moxibustion on gastric mucosal lesions may occur through neural regulation, but the precise pathways involved remain unclear. In this study,

43、 neurotomies on selected nerve tracts in rats were carried out, and the preventive effect of pre-moxibustion at acupoint Zusanli on formation of gastric mucosal lesions as well as the expression of an endogenous protective protein in serum and the gastric mucosa were observed. We aimed to explore th

44、e mechanism underlying the effect of pre-moxibustion at acupoint Zusanlim preventing gastric mucosal lesions, 2372 Peng L, et a!. /NeuralRegeneration Research. 2012;7(30):2370-2376. and determine if the pathways involved are correlated to neural regulation or humoral coordination. Based on the chang

45、es in the ulcer index, pre-moxibustion at acupoint Zusan/i has an obvious preventive effect on gastric mucosal lesions, and this effect can be interfered by neurotomy. Pre-moxibustion at acupoint Zusanlican promote heat shock protein 70 expression in serum and the gastric mucosa. The key point is th

46、at heat shock protein 70 expression in the gastric mucosa was interfered by neurotomy, but that in serum was not. This result shows that pre-moxibustion at acupoint Zusanli has good healing efficacy on gastric mucosal lesions, and that the underlying mechanism involves induction of heat shock protei

47、n 70 expression. Nerve tracts are clearly involved in the mechanism of action, so neural regulation is related to the effect of moxibustion. MATERIALS AND METHODS Design A randomized controlled animal experiment. Time and setting This experiment was conducted in the Key Laboratory of Biological Info

48、rmation Analysis for Acupuncture and Moxibustion, Hunan University of Chinese Medicine, China in 2011. Materials A total of 70 specific pathogen-free, healthy, Sprague Dawley rats aged 3-4 months and weighing 220-250 g (35 females and 35 males) were provided by the Animal Scientific Center of Hunan

49、University of Chinese Medicine. Rats were kept under the following conditions: 20-25C, humidity of 60-70%, 12-hour light/dark cycles. Experimentss were performed in direct accordance with the Guidance Suggestions for the Care and Use of Laboratory Animals, issued by the Ministry of Science and Technology of China28. Methods Transection of spinal cord After a 24-hour fast, rats were given intraperitoneal anesthesia (20% urethane, 0.6 mL/100 g), and then secured on an animal operating ta