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J. Acupunct. Tuina. Sci. 2010, 8 (1): 5-11 DOI: 10.

1007/s11726-010-0005-z

Special Topic for 973 Program

Effect of Moxibustion on Expression of HSP70 and Apoptosisrelated Factors in Rats with Acute Gastric Mucosal Damage
YI Shou-xiang ()1, YU Jie ()1, CHANG Xiao-rong ()1, PENG Yan ()1, WU Huan-gan ()2, YAN Jie ()1, YANG Ren-da ()1 1 College of Acupuncture and Massage, Hunan University of Traditional Chinese Medicine, Changsha 410007, P. R. China 2 Shanghai Research Institute of Acupuncture and Meridian, Shanghai 200030, P. R. China

HSP70 32 Wister 4 8 d Western-blotC (Cyt-c) HSP70, , Apaf-1, Caspase-9, Caspase-3, Bcl-2 Bax HSP70 , , Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 Bax (P<0.05 P<0.01) HSP70 Bcl-2 (P<0.01), Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax (P<0.01) HSP70 Bcl-2 (P<0.01), , Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax(P<0.05 P<0.01)HSP70 Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 Bax ; 70 AbstractObjective: To observe the effect of preventively used moxibustion on the expression of apoptosis-related factors in rats with acute gastric mucosal damage (GMD), and analyze the relationship between those factors and heat shock protein 70 (HSP70), for discovering the mechanism of moxibustion for protecting gastric mucosa from the aspect of mitochondria signal transduction of apoptosis. Methods: Thirty-two Wistar rats were randomly divided into binding, model, acupoint-moxibustion and non-acupoint-moxibustion groups, 8 in each group. Moxibustion was applied preventively to acupoint-moxibustion and non-acupointmoxibustion groups respectively for 8 d. Rats in binding and model groups were served with restraint without moxibustion. The GMD models were induced in groups except for A group, and the expression of cytochrome c (Cyt-c) in gastric mucosa was measured by Western-blot analysis, the expression of HSP70, cell apoptosis index (AI), apoptotic protease activating factor 1 (Apaf-1), Caspase-9, Caspase-3, Bcl-2 and Bax in gastric mucosal cells were studied by immunohistochemical detection. Results: Compared with binding group, the expressions of HSP70, AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax of gastric mucosa in model group were significantly higher (P<0.05 or P<0.01). Compared with model group, the expressions of HSP70 and Bcl-2 were significantly higher (P<0.01)while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly lower (P<0.01) in acupoint-moxibustion group.
Fund Items: National Basic Research Program of China (973 Program, 2009CB522904); National Natural Science Funds (30772707) Author: YI Shou-xiang (1946- ), female, professor Shanghai Research Institute of Acupuncture and Meridian and Springer-Verlag Berlin Heidelberg 2010 5

J. Acupunct. Tuina. Sci. 2010, 8 (1): 5-11

Compared with the acupoint-moxibustion group, the expressions of HSP70 and Bcl-2 were significantly lower (P<0.01), while AI, Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bax were significantly higher (P<0.05 or P<0.01) in non-acupoint-moxibustion group. Conclusion: Moxibustion inhibits the apoptosis of rats gastric mucosal cells and protect them from damage, through up-regulating the expression of HSP70, and modulating the expressions of Cyt-c, Apaf-1, Caspase-9, Caspase-3, Bcl-2 and Bax which were related to the mitochondria signal transduction of apoptosis. Key WordsMoxibustion; Gastric Mucosa; Apoptosis; Caspases; Mitochondria; HSP70 Heat-Shock Proteins; Rats CLC NumberR2-03 Document CodeA Our previous studies have proved that preventively used moxibustion could induce high expression of HSP70 in gastric mucosa in rats with stress gastric ulcer, to inhibit cell apoptosis and protect gastric mucosa [1, 2]. Our present study aimed to observe the effect of moxibustion on the expression of Cyt-c, Apaf-1, and the apoptosisrelated factors in the gastric mucosal cells in rats with acute gastric mucosal damage (GMD), and to discover the relationship between the aforementioned factors with HSP70, for revealing the mechanism of moxibustion for inhibiting cell apoptosis and protecting gastric mucosa from the aspect of mitochondria signal transduction of apoptosis. 1.2 Animal experiment Thirty-two healthy Wistar rats of either sex, weighing 200-250 g, aged 3-4 month, were obtained from Experimental Animal Center (Henan, China; Animal Certification Number: scxk (Yu) 20050001). The rats were randomly divided into binding group, model group, acupoint-moxibustion group C, and non-acupoint-moxibustion group, 8 in each group. Moxibustion was applied to unilateral Zusanli (ST 36) and Liangmen (ST 21)[3] or non-acupoint about 1 cm away from th location of Zusanli (ST 36) and Liangmen (ST 21) for about 30 min once everyday and continuously for 8 d. Moxa was fixed in a homemade stand and 0.5 cm apart from the acupoints or non-acupoints vertically. After fasted for 24 h, the rats in the model group, acupoint-moxibustion group and non-acupointmoxibustion group were infused with anhydrous ethanol (0.6 mL/100 g) into gastric cavity for 2 h to establish GMD model[4], while the rats in the binding group with equivalent distilled water. At the end of the experiments, gastric mucosa was sampled from randomly selected 4 rats in each group. A small piece of autral mucosa (about 5 mm10 mm) were cut from the sample and preserved in liquid nitrogen for detecting Cyt-c by Western blot method, the rest sample was preserved in paraformaldehyde (40 g/L) for detecting the expression of HSP70, Apaf-1, Caspase-9, Caspase-3, Bcl-2, Bax and cell apoptosis respectively. 1.3 Immunohistochemical detection Paraformaldehyde-fixed and paraffin-embedded tissue blocks were sectioned at 4 m, dewaxed in xylene and rehydrated through graded ethanol. Sections were treated with H2O2 30 mL/L to inhibit endogenous peroxidase activity. The expressions of

1 Material and Methods


1.1 Material Moxa stick (454 mg, 18 mm 200 mm) were purchased from Nanyang Wolong Hanyi Moxa Factory (Henan, China). HSP70 polyclonal antibody, Caspase-9 antibody, Caspase-3 antibody, Apaf-1 antibody, Cyt-c antibody were obtained from NeoMarkers Co. Ltd. Apoptosis package, 0.1mol/L PBS, TBS buffer were purchased from Wuhan Boster Biological Technology Co. Ltd (Hubei, China). Rabbit SP kit, mouse SP kit, brown DAB color-developing agent, Poly-l-lysin were purchased from Zhongshan Golden Bridge Biotechnology Co. Ltd (Beijing, China). Cell and tissue protein extraction reagent KC-415, BCA protein assay kit KC-430, biotinylated protein ladder detection pack KC-410, and KCTM chemiluminescent pack KC-420 were obtained from KangChen Bio-Tech (Shanghai, China). All the suction heads and containers are sterilized by high temperature and high pressure.

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J. Acupunct. Tuina. Sci. 2010, 8 (1): 5-11

Apaf-1, Caspase-9, Caspase-3, Bcl-2, Bax and HSP70 were detected using SP immunohistochemistry (SP kit, Zhongshan Golden Bridge Biotechnology Co. Ltd, Beijing, China). Cell apoptosis was examined using TUNEL (terminal

Differences were considered to be significant when P value was less than 0.05.

2 Results
2.1 HSP70, Apaf-1, Caspase-9, Caspase-3, Bcl-2, Bax and AI expressions in gastric mucosal cells The expressions of HSP70, Apaf-1, Caspase-9, Caspase-3, Bcl-2, Bax and AI were higher in the model group than in the binding group (P<0.05 or P<0.01). The expressions of HSP70 and Bcl-2 were higher in the acupoint-moxibustion than in the model group (P<0.01), while the expressions of Apaf-1, Caspase-9, Caspase-3, Bax and AI were significantly lower in the acupoint-moxibustion group (P<0.01). The expressions of HSP70 and Bcl-2 were lower in the non-acupoint-moxibustion group than in the acupoint-moxibustion group (P<0.01), while the expressions of Apaf-1, Caspase-9, Caspase-3, Bax and AI were significantly higher in the non-acupointmoxibustion group (P<0.05 or P<0.01). It indicated that moxibustion used as preventive treatment at Zusanli (ST 36) and Liangmen (ST 21) could increase the expressions of HSP70 and Bcl-2 and inhibit the expressions of Apaf-1, Caspase-9, Caspase-3, Bax and AI (table 1 and Fig.1).

deoxynucleotidyl transferase dUTP nick end labeling) staining (apoptosis package, Wuhan
Boster Biological Technology Co. Ltd, Hubei, China). Sections were stained with brown DAB color-developing agent. Cells bearing a brownyellow stain in the nuclei or cytoplasm were considered positive for HSP70. PCNA and cell apoptosis were considered positive when cells had a brown-yellow staining in the nuclei. Medical Image Analyzing System (MIAS) was used for object count. Control method: Negative control slides were processed with PBS liquid instead of the primary antibody but included all other steps of the procedure. 1.4 Determination of Cyt-c Expression of Cyt-c was detected by Western blot analysis. 1.5 Statistical analysis All results were expressed by mean standard deviation ( x s). Statistical significance of differences was performed by employing the ANOVA with SPSS software (version 11.5).

Table 1. Effect of moxibustion on the expressions of HSP70, Apaf-1, Caspase-9, Caspase-3, bcl-2, Bax and AI in rats with GMD 10-4 /m2, x s Groups Binding Model Acupoint-moxibustion n HSP70 AI 2.250.53 3.520.772) 3.060.81
1)6)

Apaf-1 3.440.86 4.320.671) 3.160.66


4)5)

Caspase-9 3.760.82

Caspase-3 2.350.71

Bcl-2 1.440.31

Bax 1.590.40 4.161.002) 2.520.771)4) 3.400.812)5)

8 2.510.22 8 2.930.291) 8 3.930.362)4)


2) 3) 6)

4.051.012) 3.550.862) 2.250.451) 3.330.76


5)

1.530.451)4) 2.240.492)4) 2.430.734) 1.970.614) 4.071.022)4) 2.980.86


5)

Non-acupoint-moxibustion 8 3.350.34

3.070.67

2)3)6)

Note: Compared with the binding group, 1) P<0.052) P<0.01; compared with the model group, 3)P<0.054)P<0.01; compared with the acupoint-moxibustion group, 5)P<0.056) P<0.01

Binding group

Model group

Acupoint-moxibustion group

Non-acupoint-moxibustion group

Fig.1. Effect of moxibustion on expression of HSP70 Shanghai Research Institute of Acupuncture and Meridian and Springer-Verlag Berlin Heidelberg 2010 7

J. Acupunct. Tuina. Sci. 2010, 8 (1): 5-11

2.2 Expression of Cyt-c in gastric mucosa The expression of Cyt-c was sogmofocantly higher in the model group than in the binding group (P<0.01). It was significantly lower in the acupointmoxibustion group than in the model group (P<0.01), and also significantly lower than in the non-acupoint-moxibustion group (P<0.05). It implied that infusing anhydrous ethanol into gastric cavity could increase the expression of Cyt-c in cytoplasm of gastric mucosal cells, while moxibustion performed preventively at Zusanli
1 20KD 2 3 4 1 2 3 4

(ST 36) and Liangmen (ST 21) could suppress it. (table 2 and Fig.2).
Table 2. Effect of moxibustion on the expression of Cyt-c in rats with GMDCyt-c/B-actin, x s Groups Binding Model Acupoint-moxibustion Non-acupoint-moxibustion n 4 4 4 4 Cyt-c 0.750.23 1.630.361) 0.970.262) 1.450.293)

Note: Compared with the binding group, 1) P<0.01; compared with the model group, 2) P<0.01; compared with the acupoint- moxibustion group, 3) P<0.05 1 2 3 4 1 2 3 4

Cyt-c
10KD Binding group Model group Acupoint-moxibustion group Non-acupoint-moxibustion group

Fig.2. Protein electrophoretogram of Cyt-c

3 Discussion
In recent years, a great deal of demestic researches had discovered the effects of acupuncture and moxibustion on anti-cell-apoptosis from the perspective of cerebral ischemia[5, 6], spinal cord injury[7], myocardial ischemia reperfusion[8, 9], GMD[10], ulcerative colitis[11], thymocyte and T cells in RA rats with traumatic stress[12], and proved that acupuncture and moxibustion could up-regulate the expression of HSP[1, 2]. However, which signal transduction pathway is related to the molecular mechanism of anti-apoptosis effect of HSP70 induced by acupuncture and moxibustion has not been reported up to now. Studies about the main signal pathway of cell apoptosis showed that in most cases, extracellular stimulating factors acted on cells and transformed into apoptotic signal to induce cell apoptosis by various intracellular signal transduction pathways. At present, it is considered that there are two primary signal transduction pathways: death receptor signal transduction pathway and mitochondrion signal transduction pathway[13-15]. Mitochondrion was proposed for the first time in the

late 1990s[16-18]. Nowadays, it has been proved that some apoptotic stimulating factors, such as ray, chemotherapy and medicine, by some unknown pathways, could open the permeability transition pore (PTP) and promote mitochondrion to release initiation factors (Cyt-c, AIF, Apaf-1) and Procaspase-3. The above-mentioned factors could cause cell apoptosis through the following pathways: 1) With the presence of dATP, Cyt-c could combine with Apaf-1 to show up the CARD caspase activation and recruitment domainmotif of Apaf-1 and combine with Procaspase-9 to form apoptosome. Then activated Procaspase-9 could activate Caspase-3, 6, etc. to induce cell apoptosis; 2) Bax could induce the release of Cyt-c and activate Caspase-9 to start apoptosis cascade reaction, while Bcl-2 could inhibit mitochondrial pro-apoptotic proteins, such as Cyt-c and AIF, and could also combine with Apaf-1 and Caspase 9, to inhibit caspase cascade reaction to suppress cell apoptosis; 3) AIF could induce the mitochondrion to release Cyt-c which could enhance apoptosis signal, and quickly activate endonuclease; 4) Smac/Diablo might regulate cell apoptosis by blocking the effect of inhibitors of apoptosis protein (IAPs).

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Recent studies had proved HSPs relating to mitochondrion pathway[19-22]. Over expression of HSP70 could prevent decreased transition of mitochondrion permeate and mitochondria swelling to inhibit apoptosis. HSP70 could regulate downstream of apoptosis pathway released by Cyt-c, inhibit release of Cyt-c induced by heat stress from mitochondrion, combined with recruitment domain of Apaf-1, inhibit oligomeric of Apaf-1 and activation of Caspase-9, inhibit release of Cyt-c induced by hydrogen peroxide, and then inhibit activation of Caspase-3 and subsequent myocardial apoptosis. The second kind of caspases activator found in 2000 was a kind of promoting apoptosis factors released by mitochondria. It could combine with inhibitor of apoptosis proteins (IAPs) and remove its inhibiting effect on caspases, so to enhance the activity of Caspases-3, 9 and induce apoptosis. JIANG et al[23] found that heat shock pretreatment could induce high expression of HSP70 and inhibit Smac releasing from mitochondria to cytoplasm and activation of caspase. Thus cell apoptosis was inhibited. Rokutan et al[24] summarized the action sites of HSP70 for inhibiting cell apoptosis in gastric mucosal cells in the mitochondrial signal transduction pathway as follows: 1) Up-regulating the expression of HSP70 could renature and depolymerize the degenerated protein to inhibit apoptosis; (2) HSP70 could interfere the conduction of activating signal of c-jun n-terminal kinase to inhibit apoptosis; (3) increased expression of HSP70 could prevent the reduction of mitochondrial resting membrane potential and spillover of Cyt-c and AIF, which was the dominant way to inhibit apoptosis; (4) HSP70 could combine with spillovered Cyt-c and AIF to block their combination with Caspase-9 and the conduction of apoptosis signal; (5) some of HSP70s could act with asparaginase as pseudosubstrate to prevent it from starting apoptosis procedure. The latter three points were related to mitochondria pathway. The previous studies showed that acupuncture and moxibustion had protective effect on gastric mucosa[25, 26]. Our research team had studied for a long period about the protective effect of acupunture and moxibustion performed at Foot Yangming Meridians on gastric mucosa. The mechanism had

been explored from the following aspects[27-32]: the effect of acupuncture and moxibustion on gastric mucosal blood flow and endogenous protective factors (PGE2, NO, etc), synthesis and release of brain-gut peptide (somatostatin, epidermal growth factor, transforming growth factor, substance P, etc), regulating effect on cell apoptosis of gastric mucosa and intestinal trefoil factor. Moxibustion at Zusanli (ST 36) could induce high expression of HSP70, stimulate proliferation of gastric mucosal cell and inhibit cell apoptosis to protect gastric mucosa from stress ulcer[11,33, 34]. But there were few reports about the signal transduction pathway and its regulating factors related to gastric mucosal cell apoptosis inhibited by HSP70 induced by moxibustion. Based on the above studies, it could be conferred that moxibustion could up-regulate the expression of HSP70 and interfere different targets on the apoptosis mitochondrial signal transduction pathway to protect gastric mucosa. So the present study tried to observe the effect of moxibustion used preventively on HSP70 and cell apoptosis of gastric mucosa suffered from acute damage, and explore its molecular mechanism, to reveal the interaction among moxibustion, HSP, mitochondrial signal transduction pathway and cell apoptosis. The results of this study indicated that infusing anhydrous ethanol into gastric cavity could cause the apoptosis of gastric mucosal cell. AI was higher in the model group than in the binding group, and the expressions of Bcl-2, Bax, Cyt-c, Apaf-1, Caspase-9 and Caspase-3 were also higher. it suggested that in model rats, cell apoptosis of gastric mucosa was activated by the signal pathway mediated by mitochondria and then the irreversible apoptosis process started up by the cascade reaction chain (Cyt-c Apaf-1 Caspase-9 Caspase-3). After moxibustion at acupoints, the expressions of HSP70 and Bcl-2 in gastric mucosa increased while the expressions of Bax and AI decreased. It implied that moxibustion at Zusanli (ST 36) and Liangmen (ST 21) could up-regulate the expressions of Bcl-2 and HSP70, and down-regulate the expression of Bax in gastric mucosa cells and inhibit the expressions of Cyt-c, Apaf-1, Caspase-9 and Caspase-3 which were closely related to mitochondrial signal pathway of apoptosis. Thus cell apoptosis induced by the cascade reaction chain

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(Cyt-c Apaf-1 Caspase-9 Caspase-3) decreased. Compared with the model group, the expressions of HSP70 and Bcl-2 in non-acupoint-moxibustion group were higher while the expressions of Bax and AI were lower. The expressions of Cyt-c, Apaf-1, Caspase-9 and Caspase-3 in non-acupointmoxibustion group were lower than that in the model group, but higher than that in the acupoint-moxibustion group. It suggested that, compared with performing moxibustion at nonacupoint, the regulating effect of moxibustion at acupoints on the factors related to apoptosis mitochondrial signal conduction pathway was more obvious. The results demonstrated that the effect of moxibustion on the expression of HSP70 and apoptosis was related to the comprehensive effect of moxibustion and acupoints[35]. Generally speaking, moxibustion preventively used at Zusanli (ST 36) and Liangmen (ST 21) could up-regulate the expression of HSP70 in gastric mucosa of rats with GMD, inhibit the expression of Cyt-c, Apaf-1, Caspase-9 and Caspase-3 which were related to mitochondrial signal conduction pathway to inhibit the cell apoptosis. Moxibustion might up-regulate the expression of Bcl-2 and HSP70, down-regulate the expression of Bax, so as to interfere various targets of mitochondrial signal conduction pathway to inhibit cell apoptosis and protect gastric mucosa, which was related to the comprehensive effect of moxibustion and acupoints.

References
[1] YI Shou-xiang, PENG Yan, CHANG Xiao-rong, et al. Effects of Moxibustion on Proliferation and Apoptosis of Gastric Mucosal Cells and Their Relationship with Heat Shock Protein Expression in Stress Gastric Ulcer Rats. Acupuncture Research, 2006, 31(5): 259-271. [2] Yi SX, Peng Y, Chang XR, et al. Effect of Premoxibustion on Apoptosis and Proliferation of Gastric Mucosa Cells. World J Gastroenterol, 2007, 13(15): 2174-2178. [3] LI Zhong-ren. Experimental Acupuncture Science. Beijing: China Press of Traditional Chinese Medicine, 2007: 314-329. [4] Takano H, Satoh M, Shimada A, et al. Cytoprotection by Metallothionein against Gastroduodenal Mucosal Injury Caused by Ethanol in Mice. Lab Invest 2000, 80(3): 371-377. [5] JIANG Guo-hua, TANG Wei, SUN Zhong-ren. Effect of Acupuncture to Neur Apoptosis of Mice after Full Cerebral

Ischemia. Acta Chinese Medicine and Pharmacology, 2003, 31(6): 31-32. [6] ZHOU Li, ZHENG Guo-qing. Advances of Study on Protective Effects of Acupuncture on Cerebral Ischemic Injury at Molecular Level. Chinese Acupuncture & Moxibustion, 2003, 23(5): 303-305. [7] LIANG Wen-ming, WU Ren-chang, LI Gang,etc. Effects of Electroacupuncture on Expressions of Fas and Caspase-3 in Rats with Acute Spinal Cord Injury. Chinese Journal of Clinical Rehabilitation, 2006, 10(23): 111-113. [8] TIAN Yue-feng, WU Fu-dong, QIAO Hai-fa, et al. Effect of Acupuncture of Points of the Hand-Jueyin Channel on Apoptosis in Myocardial Ischemia Reperfusion Injury. Chinese Acupuncture & Moxibustion, 2003, 23(11): 685-687. [9] ZHANG Hong-xing, ZHOU Li, HUANG Guo-fu, et al. Effect of Electroacupuncture against Apoptosis of Myocardial Ischemia Reperfusion Injury in Rabbits. Chinese Journal of Rehabilitation, 2006, 21(4): 222-224. [10] ZHOU Guo-ping, YAN Jie, LI Jiang-shan, et al. The Influence of Electro - acupuncture to Zusanli Point on Apoptosis and Regulation of Protein Experession of Bcl-2 and Fas genes in Rats with Damage of Gastric Mucosa. New Journal of Traditional Chinese Medicine, 2005, 37(4): 92-94. [11] WU Huan-gan, HUANG Zhen, LIU Hui-rong, et al. Experimental Study on Influence of Acupuncture and Moxibustion Therapy on Apoptosis of Colonic Epithelial Cells in Rats of Ulcerative Colitis. Chinese Acupuncture & Moxibustion, 2005, 25(2): 119-122. [12] WANG Jun, SUN Jing, WANG Yan-qing, et al. Effect of Electroacupuncture on Thymocyte Proliferation and Apoptosis in Surgical Trauma stress Rats. Acupuncture Research, 2006, 31(1): 12-14, 26. [13] Hengartner MO. The Biochemistry of Apoptosis. Nature, 2000, 407(6805): 770-776. [14] WANG De-cheng, GAO Hong. The Advancement on the Signal Transduction Pathways and Manipulation of Apoptosis. Progress in Veterinary Medicine, 2003, 24(6): 4-7. [15] YANG Shao-jie, MENG Jin-ping, QU Wei, et al. The Progress on the Signal Transduction Pathways of Apoptosis. Chinese Journal of Comparative Medicine. 2007, 17(5): 297-301. [16] Cui J, Chen C, Lu H, et al. Modelling of the mitochondrial Apoptosis Network. Int J Bioinform Res Appl, 2008, 4(2): 172-187. [17] Feng Y, Lu Y, Lin X, et al. Endomorphins and Morphine Limit Anoxia-reoxygenation-induced Brain Mitochondrial Dysfunction in the Mouse. Life Sci, 2008, 82(13-14): 752-763. [18] Hasenjger A, Gillissen B, Mller A, et al. Smac Induces Cytochrome c Release and Apoptosis Independently from Bax/Bcl-x(L) in a Strictly Caspase-3-dependent Manner in Human Carcinoma Cells. Oncogene, 2004, 23(26): 45234535. [19] Li CY, Lee JS, Ko YG, et al. Heat Shock Protein 70 Inhibits Apoptosis Downstream of Cytochrome C Release and Upstream of Caspase-3 Activation. J Biol Chem, 2000, 275(33): 25665-25671. [20] Saleh A, Srinivasula SM, Balkir L, et al. Negative

10 Shanghai Research Institute of Acupuncture and Meridian and Springer-Verlag Berlin Heidelberg 2010

J. Acupunct. Tuina. Sci. 2010, 8 (1): 5-11 Regulation of the Apaf-1 Apoptosome by Hsp70. Nat Cell Biol, 2000, 2(8): 476-483. [21] Ravagnan L, Gurbuxani S, Susin SA, et al. Heat-shock Protein 70 Antagonizes Apoptosis-inducing Factor. Nat Cell Biol, 2001, 3(9): 839-843. [22] XIAO Wei-min, JIANG Bi-mei, SHI Yong-zhong, et al. Mechanism of Heat Shock Protein Inhibiting C2C12 Cell Apoptosis Induced by Hydrogen Peroxide. Journal of Central South University (Medical sciences). 2004, 29(1): 6-10. [23] Jiang X, Wang X. Cytochrome c Promotes Caspase-9 Activation by Inducing Nucleutide Binding to Apaf-1. J Biol Chem, 2000, 275(40): 31199-31203. [24] Rokutan K. Role of heat shock proteins in gastric mucosal protection. J Gastroenterol Hepatol. 2000, 15(Suppl): D12-9. [25] LI Xi-ping, YAN Jie. Progress in the Research on the Protective Effect of Acupuncture and Moxibustion for Gastric Mucosal Lesion. Acupuncture Research, 2005, 30(1): 60-63. [26] YU Tian-yuan. Summarization of Studies on Protective Action of Acupuncture and Moxibustion on Gastric Mucosa. China Journal of Traditional Chinese Medicine and Pharmacy, 2003, 18(10): 626-627. [27] YAN Jie, CHANG Xiao-rong, DENG Yuan-jiang, et al. Effect of Electroacupuncture of Acupoints of Stomach Meridian on Gastric Mucosal Tissues in Alcohol-induced Gastromucosal Lesion Rabbits. Acupuncture Research, 2001, 26(4): 264-269. [28] CHANG Xiao-rong, YAN Jie, LIN Ya-jie, et al. Observation on the Cytoprotection Effect of the Gastric Mucosal Damage of the Rabbit by Electroacupuncture at Zusanli (ST36) Point. Chinese Journal of Information on Traditional Chinese Medicine, 2002, 9(7): 26-27. [29] YANG Ren-da, YI Shou-xiang, YAN Jie, et al. Study on the effect of acupuncture at Foot-Yangming Meridian acupoints on gastric mucosal damage and its mechanism. Chinese Medical Research & Clinical. 2004, 2(16): 8-11. [30] YAN Jie, LI Xi-ping, YI Shou-xiang, et al. Effect of Electroacupuncture on Gastric Mucosal Intestinal Trefoil Factor Gene Expression in the Rat of Gastric Mucosal Injury Induced by Stress. Chinese Acupuncture & Moxibustion. 2006, 26(1): 66-68. [31] YANG Zong-bao, YAN Jie. Progress of Study on the Relationship of EGFR and its Signal Transduction Ways and Recovery of Gaster Mucosa Wound. Guiding Journal of Traditional Chinese Medicine, 2006, 12(3): 66-68. [32] ZHOU Guo-ping, YAN Jie, LI Jiang-shan, et al. Effect of Electroacupuncture of Zusanli (ST 36) on Expression of Bcl-2 and Fas Proteins in Alcohol-induced Gastromucosal Lesion Rats. Acupuncture Research, 2004, 29(1): 31-34. [33] CHANG Xiao-rong, PENG Na, YI Shou-xiang, et al. Moxibustion at Zusanli and Liangmen Prevents Gastric Mucosa from Oxidation Injury through Inducing High Expression of Heat Shock Protein 70. World Chinese Journal of Digestology, 2006, 14(35): 3405-3408. [34] CHANG Xiao-rong, PENG Na, YI Shou-xiang, et al. Effects of Moxibustion on Expression of HSP70 Protein and m RNA in Stress-induced Gastric Ulcer of Rats. World Chinese Journal of Digestology, 2006, 14(13): 1252-1257. [35] YI Shou-xiang, LI Ya-ping, WU Fang, et al. Comparing Study the Protective Effect on Gastric Mucosal Lesion between Medicine Separated Moxibustion, Pure Moxibustion and Warm-up by Light at Shenque Point. Chinese Journal of Information on Traditional Chinese Medicine, 2005, 12(10): 30-32. Received Date: January 4, 2010

Introduction to Shanghai Journal of Acupuncture and Moxibustion Shanghai Journal of Acupuncture and Moxibustion (Monthly, CN 31-1317/R, ISSN 1005-0957, 64 pages) is a modern professional and academic publication on Chinese traditional acupuncturemoxibustion science, initiated in 1982, sponsored by Shanghai Academy of TCM and Shanghai Society of Acupuncture and Moxibustion, and undertaken by Shanghai Research Institute of Acupuncture and Meridian. It won excellent scientific and technologic publication prizes awarded by the Ministry of Science and Technology of the Peoples Republic of China, Science and Technology Commission of Shanghai Municipality, and Shanghai Association of Science and Technology respectively, and was selected as double-hundred periodical of Chinese Journal Phalanx in 2001. All its articles are recorded in the Chinese Journal Database, the Chinese Biomedical Journal and Literature Database. Address: No. 650, South Wanping Road, Shanghai 200030, P. R. China Telephone (Fax): 0086-021-64382181 E-mail: shzj@chinajournal.net.cn

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