Association of high expression in rat gastric mucosal heat shock protein 70 induced by moxibustion pretreatment with protection against stress injury.

AIM: To study the effect of moxibustion on Zusanli or Liangmeng point on gastric mucosa injury in stress-induced ulcer rats and its correlation with the expression of heat shock protein 70 (HSP70). METHODS: Sixty healthy SD rats (30 males, 30 females) were divided into control group, injury model group, Zushanli point group, Liangmeng point group. Stress gastric ulcer model was induced by binding cold stress method. Gastric mucosa ulcer injury (UI) index was calculated by Guth method. Gastric mucosa blood flow (GMBF) was recorded with a biological signal analyzer. Protein content and gene expression in gastric mucosal HSP70 were detected by immunohistochemistry (IHC) and reverse transcription polymerase chain reaction (RT-PCR). Thiobarbital method was used to determine malondialdehyde (MDA) content. Gastric mucosal endothelin (ET) and prostaglandin E(2) (PGE(2)) were analyzed by radioimmunoassay. RESULTS: High gastric mucosal UI index, high HSP70 expression, low GMBF and PGF(2), elevated MDA and ET were observed in gastric mucosa of rats subjected to cold stress. Moxibustion on Zusanli or Liangmeng point decreased rat gastric mucosal UI index, MDA and ET. Conversely, the expression of HSP70, GMBF, and PGE(2) was elevated in gastric mucosa after pretreatment with moxibustion on Zusanli or Liangmeng point. The observed parameters were significantly different between Zusanli and Liangmeng points. CONCLUSION: Pretreatment with moxibustion on Zusanli or Liangmeng point protects gastric mucosa against stress injury. This protection is associated with the higher expression of HSP70 mRNA and protein, leading to release of PGE(2) and inhibition of MDA and ET, impairment of gastric mucosal index.

Effect of acupuncture at Foot-Yangming Meridian on gastric mucosal blood flow, gastric motility and brain-gut peptide.

AIM: To observe the effect of acupuncture at Foot-Yangming Meridian on gastric mucosal blood flow (GMBF), gastric motility and brain-gut peptide. METHODS: Sixty SD rats were randomly divided into 6 groups: normal control group, model group (group with gastric mucosal damage, GMD), Sibai group (with acupuncture at Sibai point + GMD), Tianshu group (with acupuncture at Tianshu point + GMD), Zusanli group (with acupuncture at Zusanli point + GMD) and non-acupoint group (with acupuncture at non-acupoint + GMD). The GMD model group was induced by infusing pure alcohol into gastric cavity. H(2) Gas Clearance Test (HGCT) was used to measure GMBF, the frequency and amplitude of gastric motility were measured by the method of aerocyst, the content of brain-gut peptide in sinus ventriculi and bulbus medullae were detected by radioimmunoassay. RESULTS: Inhibitory effect of the frequency and amplitude of gastric motility were shown in model group, and the rates of frequency and amplitude changes were remarkably different from the normal control group (-19.41 +/- 17.21 vs -4.71 +/- 10.32, P < 0.05; -51.61 +/- 29.02 vs 1.81 +/- 14.12, P < 0.01). In comparison with control group, the GMBF was 0.52 +/- 0.161 mL vs 1.03 +/- 0.255 mL per 100g tissue/min, P < 0.01, the content of motilin in sinus ventriculi and bulbus medullae was 63.04 +/- 7.77 pg/mL vs 72.91 +/- 8.42 pg/mL, P < 0.05 and 50.96 +/- 8.77 pg/mL vs 60.76 +/- 8.05 pg/mL, P < 0.05, but the content of somatostatin in sinus ventriculi and bulbus medullae was 179.85 +/- 43.13 ng/g vs 90.54 +/- 40.42 ng/g, P < 0.01 and 532.86 +/- 122.58 ng/g vs 370.91 +/- 76.29 ng/g, P < 0.05,respectively. In comparison with model group, the amplitude of gastric motility was 1.52 +/- 20.13, -6.52 +/- 23.31, 6.92 +/- 25.21 vs -51.61 +/- 29.02, P < 0.01 and GMBF was 0.694 +/- 0.160 mL vs 0.893 +/- 0.210 mL, 1.038 +/- 0.301 mL vs 0.52 +/- 0.161 mL per 100g tissue/min, P < 0.01, respectively in Tianshu, Sibai and Zusanli groups, the content of motilin in sinus ventriculi and bulbus medullae was 71.64 +/- 9.35 pg/mL vs 63.04 +/- 7.77 pg/mL, P < 0.05; 58.96 +/- 6.69 pg/mL vs 50.96 +/- 8.77 pg/mL, P < 0.05 in Zusanli group and 72.84 +/- 8.62 pg/mL vs 63.04 +/- 7.77 pg/mL, P < 0.05 in Sibai group, while the content of somatostatin in bulbus medullae in Tianshu, Sibai and Zusanli groups was 480.62 +/- 151.69 ng/g, 388.53 +/- 155.04 ng/g, 365.76 +/- 143.93 ng/g vs 532.86 +/- 122.58 ng/g, P < 0.05, respectively. CONCLUSION: Electro-acupuncture at acupoints of Foot-Yangming Meridian could enhance the gastric motility, improve GMBF, and regulate the content of motilin and somatostatin in sinus ventriculi and bulbus. The effects of acupuncture on GMBF and gastric motility may be related to the content of brain-gut peptide.

Effect of pre-moxibustion on apoptosis and proliferation of gastric mucosa cells.

AIM: To observe the effects of pre-moxibustion on apoptosis and proliferation of gastric mucosal cell in rats with stress-induced ulcer, and to analyze the relationship between those effects and the expression of heat-shock protein 70 (HSP70). METHODS: Sixty healthy Sprague Dawley rats were randomly assigned into four groups, namely group A, B, C and D. The animal model of stress ulcer was established by water immersion and restraint stress. The rats in group A, B, and D served as the restraint, model, and non-acupoint controls, respectively, while those in group C received moxibustion at Zusanli and Liangmen points. Immunohistochemical methodology was used to detect the expression of HSP70, apoptosis index (AI, multiply 10(-6)/microm(2)) and proliferation index (PCNA-LI, multiply 10(-6)/microm(2)). The mucosal expression of transforming growth factor alpha(TGF-alpha) was detected by radioimmunoassay. RESULTS: Moxibustion at Zusanli and Liangmen points significantly decreased the gastric injury and the apoptosis of gastric mucosal cells, while markedly increased the mucosal expression of TGF-alpha and HSP70 as well as the proliferation of gastric mucosal cells. Compared with group A, ulcer index (UI) (26.8 +/- 9.8 vs 12.0 +/- 5.9, P < 0.01), AI (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) and expression of HSP70 (9.6 +/- 4.2 vs 4.4 +/- 2.6, P < 0.05) were significantly increased, but the content of TGF-alpha (104.7 +/- 51.2 pg/mL vs 254.0 +/- 86.9 pg/mL, P < 0.01) and PCNA-LI (6.9 +/- 4.7 vs 14.9 +/- 4.6, P < 0.05) were significantly decreased in group B. However, ulcer index values (UI) and AI were obviously lower in group C compared to groups B and D (14.1 +/- 5.4 vs 26.8 +/- 9.8 and 26.2 +/- 7.7, P < 0.01; 3.0 +/- 1.6 vs 9.6 +/- 4.2 and 8.2 +/- 5.2, P < 0.05, respectively), but content of TGF-alpha (237.0 +/- 72.6 pg/mL vs 104.7 +/- 51.2 pg/mL and 154.1 +/- 61.3 pg/mL, P < 0.01) and expression of HSP70 (0.13 +/- 0.03 vs 0.08 +/- 0.06 and 0.06 +/- 0.04, P < 0.05) were higher in group C. Furthermore, the PCNA-LI was significantly higher in group C than in group B (21.6 +/- 4.1 vs 6.9 +/- 4.7, P < 0.01). CONCLUSION: Moxibustion at Zusanli and Liangmen points has a protective effect on rats gastric mucosa in stress-induced gastric ulcer, which is closely related to its actions in promoting synthesis of TGF-alpha and proliferation of gastric mucosal cells, suppressing gastric mucosal cell apoptosis, and up-regulating HSP70 expression.

[Expressions of epidermal growth factor receptor signaling substances in gastric mucosal cells influenced by serum derived from rats treated with electroacupuncture at stomach meridian acupoints].

OBJECTIVE: To study the effects of serum derived from rats treated with electroacupuncture at stomach meridian acupoints on the expressions of epidermal growth factor receptor (EGFR) signaling substances phospholipase C gamma-1 (PLC gamma-1), protein kinase C (PKC) and c-myc in gastric mucosal cells. METHODS: Sixty rats were randomly divided into normal group, stomach meridian group, gallbladder meridian group, stomach meridian plus PD153035 group and gallbladder meridian plus PD153035 group. Water-immersion and restrained stress methods were adopted for inducing gastric mucosal injury in the rats. Gastric mucosal cells were separated by using pronase digestion method, and incubated by PD153035, a EGFR inhibitor, and 100 ml/L serum. The expression of PLC gamma-1 in the gastric mucosal cells was tested by enzyme linked-immunosorbent assay (ELISA), while the expression of PKC by isotope incorporate assay and the expression of c-myc by reverse transcription polymerase chain reaction assay (RT-PCR). RESULTS: In gastric mucosal cells, weak expressions of PLC gamma-1, PKC and c-myc were seen in the normal group, and relatively strong expressions of PLC gamma-1, PKC and c-myc were seen in the stomach meridian group and the gallbladder meridian group, among which, the expressions of PLC gamma-1, PKC and c-myc in the stomach meridian group were the strongest, and there was a significant difference between the stomach meridian group and the gallbladder meridian group (P<0.01). Relative weak expressions of PLC gamma-1, PKC and c-myc were seen in the stomach meridian plus PD153035 group and the gallbladder meridian plus PD153035 group, and there was a significant difference between the stomach meridian group and the stomach meridian plus PD153035 group (P<0.01). CONCLUSIONS: The serum derived from the rats treated with electroacupuncture at stomach meridian acupoints can activate the EGFR singling pathway, and this provides an evidence for the theory of "relative particularity between meridians and viscera" in traditional Chinese medicine.

Effect of electro-acupuncture at Foot-Yangming Meridian on somatostatin and expression of somatostatin receptor genes in rabbits with gastric ulcer.

AIM: To discuss the protective effect of electroacupuncture at the Foot-Yangming Meridian on gastric mucosal lesion, somatostatin (SS) and the expression of SS receptor genes (SSR(1)mRNA ) in rabbits with gastric ulcer and to further explore the relative specificity of meridians and viscera at gene expression level. METHODS: Forty rabbits were randomly divided into control group (A), gastric ulcer model group (B), Foot-Yangming Meridian group (C), Foot-Shaoyang Meridian group (D) and Foot-Taiyang Meridian group (E). The gastric ulcer model was prepared by infusing alcohol into stomach. Groups C-E were treated with electro-acupuncture at points along the above meridians using meridian stimulating instruments for 7 d respectively. By the end of treatment, the index of gastric ulcer was determined, the amount of epidermal growth factor(EGF) and somatostatin was measured by radioimmunoassay (RIA). SS-R(1)mRNA expression in gastric mucosa was determined by RT-PCR. RESULTS: The value of EGF in model group was obviously lower (73.6+/-14.8 vs 91.3+/-14.9 pg/mL, P<0.01) than that in control group. The index of gastric ulcer, content of SS and expression of SSR1mRNA in gastric mucosa were significantly higher than those in control group (24.88+/-6.29 vs 8.50+/-2.98 scores, P<0.01; 2978.6+/-587.6 vs 1852.4+/-361.7 mIU/mL, P<0.01; 2.56+/-0.25 vs 1.04+/-0.36, P<0.01). The value of EGF in Foot-Yangming Meridian group was higher than that in model group (92.2+/-6.7 vs 73.6+/-14.8 pg/mL, P<0.01). The index of gastric ulcer, content of SS and expression of SS-R(1)mRNA in gastric mucosa were significantly lower than those in control group (10.88+/-3.23 vs 24.88+/-6.29 scores, P<0.01; 1800.2+/-488 vs 2978.6+/-587.6 mIU/mL, P<0.01; 1.07+/-0.08 vs 2.56+/-0.25 mIU/mL, P<0.01). Compared to the model group, the content of SS and expression of SSR1mRNA in gastric mucosa in Foot-Shaoyang Meridian group decreased (2441.0+/-488.vs 2978.6+/-587.6 mIU/mL, P<0.05;1.73+/-0.16 vs 2.56+/-0.25 mIU/mL, P<0.01). But the above parameters in Foot-Taiyang Meridian group did not improve and were significantly different from those in Foot-Yangming Meridian group (P<0.05). CONCLUSION: Electro-acupuncture at Foot-Yangming Meridian can protect gastric mucosa against injury. The mechanism may be related to the regulation of brain-gut peptides and the expression of SSR(1)mRNA.

Enhanced expression of epidermal growth factor receptor gene in gastric mucosal cells by the serum derived from rats treated with electroacupuncture at stomach meridian acupoints.

AIM: To investigate the effect of serum derived from rats treated with electroacupuncture at stomach meridian acupoints on the expression of epidermal growth factor receptor (EGFR) gene in gastric mucosal cells. METHODS: The stress-induced gastric mucosal injury in rat model was established by water-immersion and restrained stress methods. 52 rats were randomly divided into: normal group (n = 8), model group (n = 8), model serum group (n = 12), stomach serum group (n = 12), and gallbladder serum group (n = 12). The gastric mucosal cells were separated by pronase-EDTA digestion method and incubated with serum. The EGFR gene expression in gastric mucosal cells was detected by reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Compared with normal group (0.6860 +/- 0.0594), the serum derived from rats of the stomach group (1.2272 +/- 0.0813, P = 0.00 < 0.01) and gallbladder group (0.9640 +/- 0.0387, P = 0.00 < 0.01) had a tendency to enhance the EGFR gene expression in gastric mucosal cells. Such tendency existed in the model group (0.7104 +/- 0.0457) but with no significant difference (P = 0.495 > 0.05) and in model serum group (0.8516 +/- 0.0409) with an extremely obvious difference (P = 0.001 < 0.01). Furthermore, the EGFR gene expression in stomach serum group was significantly higher than that in gallbladder serum group (P = 0.00 < 0.01). CONCLUSION: The present study shows that serum derived from rats treated with electroacupuncture at stomach meridian acupoints can distinctly increase the EGFR gene expression of gastric mucosal cells. Therefore, there is certain meridian specificity in the serum, which could provide a proof for the TCM theory "particular relation between meridian and internal organ".

Effect of electroacupunture on gastric mucosal intestinal trefoil factor gene expression of stress-induced gastric mucosal injury in rats.

AIM: To investigate electroacupuncture(EA) at the acupoints of Stomach Meridian of Foot-Yangming (SMFY), Gallbladder Meridian of Foot-Yangming (SMFY) on gastric mucosal intestinal trefoil factor (ITF) gene expression detection in stress-induced rats with gastric mucosal lesion, and to explore the regulatory mechanism and significance of EA-related gastric mucosal protective effect. METHODS: Forty rats were randomly divided into 4 groups: Blank group, Model group, Model group+EA at acupoints of SMFY group ("SMFY group"), and Model group+EA at acupoints of GMFY group(GMFY group). All rats (except blank group) were made model by water immersion and restraint stress (WRS). Then the gastric mucosa tissue in each rat was taken off after assessment of gastric mucosal lesion index(GUI), and the expression of ITF mRNA of the tissues was detected by reverse transcription-polymerase chain reaction (RT-PCR) method. RESULTS: Compared with Model group(54.3+/- 1.34), the GUI value in SMFY group (31+/- 2.21) decreased significantly(P< 0.01), so did that in GMFY group (39.8+/- 1.62, P< 0.05), meanwhile GUI value in SMFY group was significantly lower than in GMFY group(P< 0.01). Compared with Model group (0.65+/- 0.01), EA had a tendency to improve the expression of gastric mucosal ITFmRNA gene: such tendency existed in GMFY group (0.66+/- 0.01) but with no significant difference(P>0.05), in SMFY group(0.76+/- 0.01) with an extremely obvious difference (P< 0.01), furthermore the expression in SMFY group was significantly higher than in GMFY group (P< 0.01). CONCLUSION: The gastric mucosal protective effect by EA at the acupoints of SMFY and GMFY was related to the expression variance of ITF, indicating certain meridian specificity exists. It could be one proof for the TCM theory "Relative particularity between SMFY and stomach".

The effect of EA on the gastric mucosal histology and ITF mRNA expression in stress-induced rats.

This study focuses on the effect of electroacupuncture (EA) on the gastric mucosal histology and ITF (intestinal trefoil factor) mRNA in stress-related rat, and the relationship between the gastric protective mechanism of EA at acupoints of Stomach Meridian of Foot-Yangming (SMFY) group and Gallbladder Meridian of Foot-Shaoyang (GMFS) group. Forty rats were randomly divided into 4 groups: blank control group (BCG), model control group (MCG), SMFY group (EA at acupoints of SMFY for 7 days before model inducing), and GMFS group (EA at acupoints of GMFS for 7 days before model inducing). All rats (except normal group) were made model by water immersion and restriction (WRS) on day 7, then the gastric mucosal lesion index (GUI) was accessed, ITF mRNA expression of the tissue was detected by reverse- transcriptase-polymerase chain reaction (RT-PCR) method, and the histological change under light microscope was observed. As a result, the GUI value in SMFY/GMFS groups decreased significantly (p < 0.05 or 0.01). The level of ITF mRNA expression in SMFY group was significantly higher than that in MCG (p < 0.01), while that in GMFS group was higher than MCG but there was no statistical difference (p < 0.05). This result may be due to the intrinsic mechanism of EA's gastric mucosal protection by the upregulation of ITF mRNA expression in gastric mucosal tissue, and the expression variance indicated the classical traditional Chinese medicine (TCM) theory "Relative Particularity between SMFY and Stomach.".

Effect of acupuncture at different meridian acupoints on changes of related factors for rabbit gastric mucosal injury.

AIM: To explore the regularity of multi-meridians controlling a same viscus (MMCSV). METHODS: The rabbit gastric ulcer model was established by ethanol intragastric instillation. Fifty-six rabbits were randomly divided into normal group, model group (MG), model plus acupuncture at Foot Yangming Meridian group (YMG), model plus acupuncture at Foot Taiyin Meridian group (TYG), model plus acupuncture at Foot Shaoyang Meridian group (SYG), model plus acupuncture at Foot Jueyin Meridian group (JYG), model plus acupuncture at Foot Taiyang Meridian group (TYMG), with eight rabbits in each group. Gastric mucosal nitric oxide (NO) and nitric oxide synthase (NOS) were assayed by the nitric acid reductase method, and prostaglandin E2 (PGE2) and epidermal growth factor (EGF) were measured by radioimmunoassay. The comprehensive effects were analyzed by weighing method. RESULTS: Compared to MG, SYG, JYG and TYMG, the rabbits gastric mucosal injury index (GMII) reduced very significantly in YMG (P<0.01). Compared to MG, the GMII also reduced significantly in TYG (P<0.05). NO, NOS, PGE2 and EGF increased very significantly in YMG (P<0.01). The EGF in YMG also increased significantly than that in TYG compared to those in MG, SYG, JYG and TYMG (P<0.05). The PGE2 and EGF also increased very significantly in TYG than those in MG, JYG and TYMG (P<0.01). While compared to SYG, the NOS increased significantly in TYG (P<0.05). NOS was the highest in YMG (P<0.01), and was higher in TYG than in MG (P<0.01). CONCLUSION: MMCSV is common. The Foot Yangming Meridian is most closely related to the stomach, followed by Foot Taiyin Meridian, Foot Shaoyang Meridian and Foot Jueyin Meridian. Foot Taiyang Meridian has no correlation with the stomach.

Effects of electroacupuncture on gastric myoelectric activity and substance P in the dorsal vagal complex of rats.

Acupuncture has been widely applied to treat gastrointestinal diseases in the clinic. However, its precise mechanism remains unknown. In the present study, effects of electroacupuncture (EA) at Zusanli acupoints (ST36) on gastric myoelectric activity and substance P (SP) in the dorsal vagal complex (DVC) were observed. EA at ST36 significantly increased the frequency of slow waves and the frequency of the spike potentials superimposed on the slow waves, which persisted for 90 min after cessation of EA and was completely abolished after bilateral vagotomy. Meanwhile, the mean optical density of SP immunoreactive fibers in the DVC increased following EA. The results suggested that EA at ST36 promoted the gastric myoelectric activity, which was regulated by the vagus, and SP in the DVC may be involved in the excitatory effects. Furthermore, EA had a relative long-term post-effect. We presumed that DVC may be a supraspinal center in the effect of EA on gastric function.

[Serumimmunological study of moxibustion on helicobacter pylori gastritis in rats].

OBJECTIVE: To explore the immune mechanism of moxibustion on protecting gastric mucosa injury. METHODS: Forty healthy SD rats were randomly divided into four groups: a blank group, a model group, a moxibustion acupoint group and a moxibustion non-acupoint group, 10 rats in each one. Eight days before model establishment, moxibustion at "Zusanli" (ST 36), "Zhongwan" (CV 12), "Guanyuan" (CV 4), "Pishu" (BL 20) and "Weishu" (BL 21) was applied in the moxibustion acupoint group while these acupoints' controlled points were selected in the moxibustion non-acupoint group, and no treatment was given in the model group, once a day in three groups for continuous 16 days. The helicobacter pylori (Hp) model was established by intragastric administration of Hp. HE staining microscopic examination was used to observe inflammation severity in gastric mucosa, and enzyme-linked immunosorbent assay (ELISA) was adapted to measure content of heat shock protein (HSP) 72, TNF-alpha and IL-1beta, and real-time quantitative PCR was used to measure the expression of TLR2 mRNA, TLR4 mRNA, CD14 mRNA and MyD88 mRNA in peripheral blood mononuclear cells, and western blot method was used to measure content of NFkappaB and IkappaBalpha in peripheral blood mononuclear cells. RESULTS: Compared with the blank group, the expression of HP could be seen in the smear of gastric mucosa by Gram's staining in the model group; the inflammation severity score was obviously increased as well as content of serum HSP 72 and TNF-alpha and IL-1beta in gastric tissue; and expression of TLR2, 4 mRNA, CD14 mRNA, MyD88 mRNA, NFkappaB was increased (P < 0.01), but the expression of IkappaBalpha was reduced (P < 0.05). After the moxibustion, the inflammation severity score was reduced in the moxibustion acupoint group, and the content of serum HSP 72 was increased, and the expression of TNF-alpha and IL-1beta in gastric tissue and expression of TLR2 mRNA, TLR4 mRNA, CD14 mRNA, MyD88 mRNA and NFkappaB were reduced (P < 0.01), but the expression of IkappaBalpha was increased (P < 0.05). The differences between the moxibustion non-acupoint group and the model group were not significant (P > 0.05). CONCLUSION: The pretreatment of moxibustion at acupoints could induce the over expression of serum HSP 72. By combining TLR 2 and 4 receptors to trigger receptor signal transduction pathways, the releases of downstream signal substances are regulated; as a result, the releases of related immune substances are regulated to relieve the gastric mucosa injury of rats with HP gastritis.

[Effect of manual acupuncture stimulation of "Zusanli" (ST 36) on gastric motility, and SP and motilin activities in gastric antrum and nucleus raphe magnus in gastric hyperactivity and hypoactivity rats].

OBJECTIVE: To observe the changes of gastric motility and levels of substance P (SP) and motilin (MTL) in the gastric antrum and Nucleus Raphe Magnus (NRM) after manual acupuncture stimulation of "Zusanli" (ST 36) in gastric hyperactivity and hypoactivity rats, so as to analyze the role of NRM in acupuncture mediated adjustment of gastric motility. METHODS: Fifty SD rats were randomly and equally divided into control, gastric hyperactivity (G-Hypera) model, gastric hypoactivity (G-Hypoa) model, acupuncture + G-Hypera and acupuncture + G-Hypoa groups (10 rats/group). G-Hypera model was established by intravenous (tail vein) injection of Maxolon (0.5 mL/200 g) and G-Hypoa model established by intravenous injection of Atropin (0.5 mL/200 g), respectively. After insertion of acupuncture needles into bilateral "Zusanli" (ST 36), the needles were repeatedly manipulated at a frequency of about 2 Hz for 5 min. The intragastric pressure was recorded and analyzed using a physiological signal analysis system. The SP and MTL contents of gastric antrum were measured by ELISA, and SP and MTL immunoactivity of NRM was determined by immunohistochemistry. RESULTS: In gastric hyperactivity rats, compared with the control group, the intragastric pressure (not systolic frequency), SP and MTL contents in the gastric antrum and MTL immunoactivity of NRM were significantly increased (P < 0.05, P < 0.01), while SP immunoactivity of NRM was apparently decreased in the G-Hypera model group (P < 0.01). After acupuncture stimulation, the intragastric pressure, gastric SP and MTL contents, and MTL immunoactivity of NRM were all significantly down-regulated (P < 0.05, P < 0.01), and SP immunoactivity of NRM was markedly up-regulated (P < 0.01) in comparison with the model group. In gastric hypoactivity rats, compared with the control group, the intragastric pressure and gastric systolic frequency, gastric SP and MTL contents, and MTL immunoactivity of NRM were significantly decreased (P < 0.05, PF < 0.01), while SP immunoactivity of NRM was remarkably up-regulated in the G-Hypoa model group (P < 0.01). After acupuncture stimulation, the above-mentioned 6 indexes were reversed significantly (P < 0.05, P < 0.01) in comparison with those of the model group. CONCLUSION: Acupuncture stimulation of "Zusanli" (ST 36) has an obvious bi-directional adjustment effect on gastric motility, which is closely associated with its functions in regulating gastric SP and MTL level and the expression of MTL and SP in the NRM of brainstem.

[Clinical observation on therapeutic effects of mild moxibustion in superficial gastritis patients with spleen-stomach deficiency-cold pattern].

OBJECTIVE: To observe the curative effect of different duration of mild moxibustion in superficial gastritis patients with deficiency cold pattern of spleen and stomach. METHODS: A total of 84 superficial gastritis patients were randomly divided into moxibustion-20 min group, moxibustion-40 min group and medication (Ranitidine) group, with 28 cases in each group. Mild moxibustion was applied to bilateral Zusanli (ST 36) and Zhongwan (CV 12, odd days), Pishu (BL 20) and Weishu (BL 21, even-numbered days) for 20 min and 40 min respectively, once daily for 20 days. The symptom integrative score of superficial gastritis patients and the therapeutic effects were assessed according to "The Scoring Table for Symptoms and Signs of Gastrointestinal Disorders of Chinese Medicine", etc. RESULTS: Following 20 days' treatment, of the three 28 gastrititis patients in the medication, moxibustion-20 min and moxibustion-40 min groups, 0, 1, and 0 cases were cured, 5, 7 and 12 experienced marked improvement in their symptoms, 15, 16 and 12 had an improvement, and 8, 4 and 4 were invalid, with the effective rates being 71.4%, 85.7% and 85.7%, respectively. After one month's follow up, of the three 28 cases in the medication, moxibustion-20 min and moxibustion-40 min groups, 0, 4 and 0 cases were cured, 4, 8, and 13 experienced marked improvement, 15, 16 and 13 had an improvement, 9, 0 and 2 were ineffective, with the effective rates being 67.9%, 100% and 92.9%, respectively. Self-comparison of each group showed that the integrative scores of symptoms were all decreased significantly after the treatment and one-month's follow up in the three groups (P < 0.01). The effective rates of both moxibustion-20 min and -40 min were significantly higher than that of the medication group after one month's follow up (P < 0.01, P < 0.05). The therapeutic effect of moxibustion-20 min group was significantly superior to that of the medication group in lowering the integrative score of symptoms after one month's follow up (P < 0.01). No significant differences were found among the three groups in the therapeutic effects and in reducing integrative scores of symptoms after 20 days' treatment (P > 0.05). CONCLUSION: Mild moxibustion is significantly superior to medication in long-term therapeutic effects in the treatment of superficial gastritis patients with spleen-stomach deficiency-cold pattern.

[Effect of moxibustion on motility, absorption and content of ATP in small intestine of spleen-deficiency rats].

OBJECTIVE: To explore the mechanism of reinforcing function of moxibustion to spleen-stomach. METHODS: Forty healthy Sprague Dawley rats were randomly divided into 4 groups: group A (blank group), group B (model group), group C (moxibustion group) and group D (herbs group). The rat model of spleen-deficiency was established by intragastric administration with 200% Dahuang (Rhubarb) infusion. The rats in group A and B, and D served as the blank control, model, and Sijunzi decoction group respectively, while those in group C received moxibustion at "Zusanli" (ST 36), "Zhongwan" (CV 12), "Guanyuan" (CV 4), "Pishu" (BL 20) and "Weishu" (BL 21), etc. The common symptoms and intestinal propulsive rate were observed. The content of I-xylose in serum was detected by phloroglucinol method. Colorimetry method was used to detected content of ATP in jejunum tissues. RESULTS: Compared with group A, the symptom score in group B was increased significantly (both P < 0.01), while the intestinal propulsive rates, the content of D-xylose in serum and ATP in jejunum tissues were decreased significantly (P < 0.05, P < 0.01). Compared with group B, the symptom score in group C and D was decreased significantly (both P < 0.01), while the intestinal propulsive rates, the content of D-xylose in serum and ATP in jejunum tissues were increased significantly (P < 0.05, P < 0.01). There were no significant difference between group C and D (P > 0.05). CONCLUSION: Moxibustion at "Zusanli" (ST 36) etc. could relieve symptoms of spleen-deficiency, enhance motility and absorption functions of small intestine and improve metabolism of small intestine. The efficacy is equal to administration of Sijunzi decoction.

[Effect of plasma of healthy subjects undergoing moxibustion on ethanol-injured human gastric epithelial GES-1 cells in vitro and the involved mitochondrial apoptosis pathway].

OBJECTIVE: To observe the effect of plasma derived from healthy volunteers undergoing moxibustion (moxibustion plasma) on alchol-injured human gastric epithelial GES-1 cells in vitro, and expression of heat shock protein 70 (HSP 70, cell apoptosis inhibitory protein), apoptosis inducing factor (AIF), Smac (a mitochondrial protein), and Caspase 3 and Caspase 9 (the latter 3 proteins are also involved in cell apoptosis) in order to study its mechanisms underlying protecting gastric mucous membrane. METHODS: Twenty-four healthy volunteer subjects (half men and half women) were randomized into acupoint-moximustion (A-M) [Zhongwan(CV 12), Guanyuan (CV 4) and Zusanli (ST 36)] group and non-acupoint-moxibustion (NA-M, 3 cun right to CV 12 and CV 4.1 cun medial to ST 36 ) group (n = 12/group). Moxibustion was applied to the above-mentioned 3 acupoints and non-acupoints for 30 min, once daily for 10 days. Venous blood of the subjects was collected before and after moxibustion. The cultured GES-1 cells were divided into: control group. ethanol-injury group (model), A-M plasma group (A-M-P, plasma got from volunteers undergoing A-M), and NA-M plasma group (NA-M-P,plasma got from volunteers accepting NA-M). The GES-1 cells of the latter 3 groups were treated with 8% ethanol for duplicating cell injury model. Apoptosis was detected by flowcytometry. Expression of HSP 70, second mitochondria-derived activator of Caspase (Smac) and AIF proteins of GES-1 cells were assayed by western blotting, and the immunoactivity of cysteinyl aspirate-specific proteinase-3 and 9 (Caspase-3, 9) was detected by immunocytochemistry. RESULTS: In comparison with the control group, the apoptosis rate, the expression of HSP 70, Smac and AIF proteins, and the immunoactivity of Caspase-3 and Caspase-9 of the model group were increased significantly (P < 0.01). Compared with the model group, the apoptosis rate of GES-1 cells, the expression of Smac and AIF proteins, and the immunoactivity of Caspase-3 and Caspase-9 in the A-M-P group, the apoptosis rate, the expression of Smac and the immunoactivity of Caspase-3 and Caspase-9 in the NA-M-P group were all down-regulated considerably (P < 0.05, P < 0.01). In comparison with the model group, HSP 70 expression of the A-M-P group was up-regulated significantly (P < 0.01). The apoptosis rate of GES-1 cells, the expression levels of Smac, AIF, Caspase-3 and Caspase-9 were significantly lower in the A-M-P group than in the NA-M-P group (P < 0.05, P < 0.01), while the expression of HSP 70 was apparently higher in the A-M-P group than in the NA-M-P group (P < 0.01). CONCLUSION: Plasma derived from the subjects undergoing moxibustion of Zusanli (ST 36), Zhongwan (CV 12) and Guanyuan (CV 4) can inhibit apoptosis of GES-1 cells in vitro, which is closely related to its effects in up-regulating intracellular HSP 70 expression and down-regulating mitochondrial apoptosis protein expression of AIF. Smac, Caspase-3 and Caspase-9.

[Effects of moxibustion on gastric mucosal EGF and TGF-alpha contents and epidermal growth factor receptor expression in rats with gastric mucosal lesion].

OBJECTIVE: To observe the effects of moxibustion on contents of epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) and epidermal growth factor receptor (EGFR) expression in the gastric mucosa tissue in rats with gastric mucosal lesion. METHODS: Thirty-six SD rats were randomly and equally divided into control, model and moxibustion groups. Gastric mucosal lesion model was duplicated by restraint and cool water immersion stress. Pre-moxibustion was applied to "Zusanli" (ST 36) and "Zhongwan" (CV 12), "Pishu" (BL 20) and "Weishu" (BL 21) alternately, once everyday for 8 days before modeling. The contents of EGF and TGF-alpha in gastric mucosa were detected by enzyme-linked immunosorbent assay (ELISA) and the expression of EGFR determined by immunohistochemistry. RESULTS: Compared with the control group, only TGF-alpha content in the gastric mucosa in the model group was increased significantly (P < 0.05). Compared with the model group, the EGF and TGF-alpha contents and EGFR immunoactivity in the gastric mucosa were increased significantly in the moxibustion group (P < 0.05, P < 0.01). CONCLUSION: Pre-moxibustion at ST 36, CV 12, BL 20 and BL 21 can up-regulate gastric mucosal EGF and TGF-alpha contents and EGFR protein expression in gastric mucosa lesion rats, which may contribute to its effect in relieving stress-induced gastric mucosal injury.

[Progress of studies on acu-moxibustion stimulation-induced cellular transmembrane signal transduction of the target-organs].

Abundant research results have shown that multiple levels and links of cellular transmembrane signal transduction pathways in the target organs were involved in the efficacy of acupuncture. For instance, 1) various extra-cellular growth factors for initiating signal transduction by activating tyrosine protein kinase and non-receptor tyrosine kinase, 2) G protein-coupled protein kinase-second signal messengers, 3) ligands acting on intra-nuclear receptors to activate transduction pathway of nuclear transcription factors of the target genes, have been demonstrated in the favorable regulating process of acupuncture and moxibustion in different pathological animal models. In the present paper, the authors review the progress of studies on the abovementioned mechanism of acu-moxibustion underlying improving some disorders as 1) pain, cerebral ischemia, and senile dementia, 2) inflammation and tumor, and 3) myocardial ischemia. Moreover, the authors also analyze the extant problems and make a prospect on the future studies about the cellular transmembrane signal transduction pathways involving the effects of acupuncture and moxibustion.

[Effect of different frequency electroacupuncture on the expression of substance P and beta-endorphin in the hypothalamus in rats with gastric distension-induced pain].

OBJECTIVE: To observe the effect of different frequency electroacupuncture (EA) on hippocampal substance P (SP) and beta-endorphin (beta-EP) in rats with gastralgia so as to explore its underlying mechanism in preventing and relieving visceral pain. METHOD: Fifty rats were randomly divided into control, sham operation, model, low- frequency-EA and high frequency-EA groups. Gastralgia model was duplicated by gastric distension (a pressure of 40 mmHg) with a latex balloon. Low (2 Hz) and high (100 Hz) frequency EA was applied to bilateral "Zusanli (ST 36)" for 30 min. Pain score was given by using Al-Chaer's method (abdominal withdrawal reflex) and the expression of hypothalamic beta-EP and SP was detected by immunohistochemistry. RESULTS: Both low- and high-frequency EA of ST 36 could significantly lower the pain score (P < 0.01), and the effect of high-frequency EA was markedly superior to that of low-frequency (P < 0.01). Compared with control and sham-operation groups, both SP immuno-reaction (IR) and beta-EP IR positive cells in the hypothalamus in model group increased significantly in the number (P < 0.01), suggesting an upregulation of SP and beta-EP expression after occurrence of gastralgia. In comparison with model group, the number of SP IR and (beta-EP IR positive cells in the hypothalamus in low frequency and high frequency EA groups increased further (P < 0.01) particularly in low frequency EA group. CONCLUSION: Both low-frequency and high-frequency EA of "Zusanli (ST 36)" can effectively relieve gastric distension-induced gastralgia in the rat, and the effect of high-frequency EA is better, which may be closely related to its actions in promoting the release and synthesis of hypothalamic beta-EP and SP.

[Observation on therapeutic effect of opposing needling for treatment of poststroke shoulder-hand syndrome].

OBJECTIVE: To compare the therapeutic effects of opposing needling and routine acupuncture for treatment of poststroke shoulder-hand syndrome. METHODS: Sixty cases were randomly divided into an opposing needling group and a routine acupuncture group, 30 cases in each group. In the two groups, Jianyu (LI 15), Jianliao (TE 14), Quchi (LI 11), Hegu (LI 4), etc. were selected, with on the healthy side selected for the opposing needling group and on the affected side selected for the routine acupuncture group. The motor function of the affected limb was evaluated by Fugl-Meyer scale, activity of daily living by ADL scale, and pain by VAS, and the edema degree was investigated before and after treatment. RESULTS: There were significant differences before treatment and after 2 therapeutic courses in the scores of Fugl-Meyer for the upper limb motor function and the ADL score for activity of daily living (both P<0.05), with the opposing needling group being significantly better than the routine acupuncture group (P<0.05); after treatment, both the edema degree and the pain scores significantly decreased (both P<0.05), with the opposing needling group in improvement of the edema degree being better the routine acupuncture group (P<0.05), and with no significant difference between the two groups in improvement of pain (P>0.05); the total effective rate was 93.3% in the opposing needling group and 90.0% in the routine acupuncture group with a significant difference between the two groups (P<0.05). CONCLUSION: The clinical therapeutic effect of the opposing needling is better than that of routine acupuncture therapy for treatment of poststroke shoulder hand syndrome.

[Observation on changes of the intragastric protein phosphorylation level involving electroacupuncture-induced improvement of the injured gastric mucosa in the rat].

OBJECTIVE: To study the effect of electroacupuncture (EA) of Stomach Meridian acupoints on changes o of the related protein phosphorylation in the gastric mucosa involving the repair of the injured gastric mucosa in the rat. METHODS: A total of 20 SD rats were divided into control, model, acupoint and non-acupoint groups, with 5 cases in each. Gastric mucosa injury model was established by intragastric perfusion of dehydrated alcohol (0.8 ml/100 g). EA (4 Hz/50 Hz, 1-3 mA) was applied to "Zusanli" (ST 36), "Liangmen" (ST 21) and "Sibai" (ST 2), and the corresponding non-acupoints, once daily for 7 days. Gastric mucosal ulcer index (UI) was measured, and the profiling of the protein phosphorylation of interest in gastric mucosa cells was detected by antibody microarrays. RESULTS: In comparison with control group, the gastric mucosal UI of model group was significantly higher than that of control group (P < 0.01), while the UI of EA group was significant lower than those of model group and non-acupoint group (P < 0.01), suggesting a recovery of the injured mucous membrane. Findings of protein array analysis indicated that a total of 720 kinds of phosphorylated proteins (> or = 1.5 folds) were found in the injured gastric mucosa. Compared to model group, the phosphorylation levels of 100 and 20 proteins were up-regulated, and 16 and 9 proteins down-regulated in acupoint and non-acupoint groups, respectively. The up-regulated proteins involve cell cycle mediation, cell proliferation, anti-apoptosis, and the regulation of the cell adhesion, cytoskeleton protein, inflammation, immune activities, etc. while the down-regulated proteins involve the down-regulation of the phosphorylation levels of the apoptosis proteins, adhesion proteins and cytoskeleton proteins, etc. In comparison to non-acupoint group, the phosphorylation levels of 100 proteins were up-regulated and 16 proteins down-regulated in acupoint group. CONCLUSION: EA of acupoints of the Stomach Meridian can promote repair of the injured gastric mucosa, which may be related to its effects in regulating the levels of phosphorylation of many signaling proteins.