Effects of moxibustion pretreatment on extracellular signal-regulated kinase signaling transduction pathway in the gastric tissues of rats with gastric mucosal damage

Objective： To observe the effects of moxibustion pretreatment on the protein expressions of epidermal growth factor receptor （EGFR）, phosphorylation extracellular signal-regulated kinase I/2 （p-ERKI/2） and activated protein-1 （AP-2）, the key factors of extracellular signal-regulated kinase signaling transduction pathway in gastric tissue of rats with stress-induced gastric mucosal damage, and to discuss the mechanisms of moxibustion therapy in promoting the restoration of damaged gastric mucosa. Methods： Thirty Sprague-Dawley （SD） rats were randomly divided into a normal group, a model group, and a moxibustion group using the random digits table, 10 in each group. Except the rats in the normal group, rats in the other two groups were used to make stress-induced gastric mucosal damage model using restraint and cold stress. Before modeling, rats in the moxibustion group were alternately treated with moxibustion a/t Zusanli （ST 36） and Zhongwan （CV 12）, or Pishu （BL 20） and Weishu （BL 22）, once a day, for a total of 8 d. Histolo^cal changes of gastric mucosa were observed under the light microscopy, the expression of gastric tissue p-ERKI/2 was detected by immunohistochemistry assay, and the protein levels of EGFR and AP-I were measured by Western blots. Results： Compared with rats in the normal group, gastric mucosal damage was more serious, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the model group （P〈0.01, P〈O.05, P〈0.05）. Compared with rats in the model group, gastric mucosal damage was milder, and protein expressions of gastric tissue EGFR, p-ERK1/2 and AP-1 increased in the moxibustion group （all P〈0.01）. Conclusion： Moxibustion at Zusanli （ST 36）, Zhongwan （CV 12）, Pishu （BL 20） and Weishu （BL 21） could increase EGFR, p-ERK1/2 and AP-1 expression levels in gastric tissue of stress-induced gastric mucosal damage rats, maintain the information transfer function of ERK signaling transduction pathway, and promote restoration of gastric mucosal damage.

Organic carbon monoxide prodrug, BW-CO-111, in protection against chemically-induced gastric mucosal damage

Metal-based carbon monoxide(CO)-releasing molecules have been shown to exert antiinflammatory and anti-oxidative properties maintaining gastric mucosal integrity.We are interested in further development of metal-free CO-based therapeutics for oral administration.Thus,we examine the protective effect of representative CO prodrug,BW-CO-111.in rat models of gastric damage induced by necrotic ethanol or aspirin,a representative non-steroidal anti-inflammatory drug.Treatment effectiveness was assessed by measuring the microscopic/macroscopic gastric damage area and gastric blood flow by laser flowmetry.Gastric mucosal mRNA and/or protein expressions of HMOX1,HMOX2,nuclear factor erythroid 2-related factor 2,COX1,COX2,iNos,Anxa1 and serum contents of TGFB1,TGFB2,IL1 B,IL2,IL4,IL5,IL6,IL10,IL12,tumor necrosis factorα,interferonγ,and GM-CSF were determined.CO content in gastric mucosa was assessed by gas chromatography.Pretreatment with BW-CO-111(0.1 mg/kg,i.g.)increased gastric mucosal content of CO and reduced gastric lesions area in both models followed by increased GBF.These protective effects of the CO prodrug were supported by changes in expressions of molecular biomarkers.However,because the pathomechanisms of gastric damage differ between topical administration of ethanol and aspirin,the possible protective and anti-inflammatory mechanisms of BW-CO-111 may be somewhat different in these models.

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

AIM： To investigate electroacupunture（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 alter assessment of gastric mucosal lesion index（GUI）, and the expression of ITF mRNA of the tissues was detected by reverse transcdption-polymerase chain reaction（RT-PCR） method. RESULTS： Compared with Model group（S4.3± 1.34）, the GUI value in SMFY group （31±2.211 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 signficant 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 pardcularity between SMFY and stomach＂.

Ghrelin attenuates gastrointestinal epithelial damage induced by doxorubicin

AIM:To examine the influence of ghrelin on the regenerative potential of gastrointestinal(GI)epithelium.METHODS:Damage to GI epithelium was induced in mice by two intravenous injections of doxorubicin(10 and 6 mg/kg).Some of the doxorubicin-treated mice received a continuous subcutaneous infusion of ghrelin(1.25μg/h)for 10 d via implanted mini-osmotic pumps.To label dividing stem cells in the S-phase of the cell cycle,all mice received a single intraperitoneal injection of 5'-bromo-2'-deoxyuridine(BrdU)one hour before sacrifice.The stomach along with the duodenum were then removed and processed for histological examination and immunohistochemistry using anti-BrdU antibody.RESULTS:The results showed dramatic damage to the GI epithelium 3 d after administration of chemotherapy which began to recover by day 10.In ghrelintreated mice,attenuation of GI mucosal damage was evident in the tissues examined postchemotherapy.Immunohistochemical analysis showed an increase in the number of BrdUlabeled cells and an alteration in their distribution along the epithelial lining in response to damage by doxorubicin.In mice treated with both doxorubicin and ghrelin,the number of BrdUlabeled cells was reduced when compared with mice treated with doxorubicin alone.CONCLUSION:The present study suggests that ghrelin enhances the regenerative potential of the GI epithelium in doxorubicintreated mice,at least in part,by modulating cell proliferation.

Effect of lysozyme chloride on betel quid chewing aggravated gastric oxidative stress and hemorrhagic ulcer in diabetic rats

AIM： To evaluate the protective effect of lysozyme chloride on betel quid chewing （BQC） aggravated gastric oxidative stress and hemorrhagic ulcer in rats with diabetes mellitus （DM）. METHODS： Male Wistar rats were challenged intravenously with streptozotocin （65 mg/kg） to induce DM. Rats were fed with regular pellet food or BQC-containing diets. Afcer 90 d, rats were deprived of food for 24 h. Rat stomachs were irrigated for 3 h with normal saline or simulated gastric juice. Rats were killed and gastric specimens were harvested. RESULTS： An enhancement of various gastric ulcerogenic parameters, including acid back-diffusion, mucosal lipid peroxide generation, as well as decreased glutathione levels and mucus content, were observed in DM rats. Afcer feeding DM rats with BQC, an exacerbation of these ulcerogenic parameters was achieved. Gastric juice caused a further aggravation of these ulcerogenic parameters. Daily intragastric lysozyme chloride dose-dependently inhibited exacerbation of various ulcerogenic parameters in those BQC-fed DM rats. CONCLUSION： （1） Gastric juice could aggravate both DM and BQC-fed DM rat hemorrhagic ulcer; （2） BQC exacerbated gastric hemorrhagic ulcer in DM rats via enhancing oxidative stress and reducing defensive factors; （3） lysozyme chloride effectively protected BQC aggravated gastric damage in DM rats.

Effect of ecoimmunonutrition supports on maintenance of integrity of intestinal mucosal barrier in severe acute pancreatitis in dogs

Background One of the major causes of death in severe acute pancreatitis （SAP） is severe infection owing to bacterial translocation. Some clinical studies suggested that ecoimmunonutrition （EIN） as a new strategy had better treatment effect on SAP patients. But the experiment studies on the precise mechanism of the effect of EIN were less reported. In this study, we mainly investigated the effects of EIN on bacterial translocation in SAP model of dogs. Methods SAP was induced by retrograde infusion of 5% sodium taurocholate into the pancreatic duct in healthy hybrid dogs. The SAP dogs were supported with either parenteral nutrition （PN） or elemental enteral nutrition （EEN） or EIN. The levels of serum amylase, serum aminotransferase and plasma endotoxin were detected before and after pancreatitis induction. On the 7th day after nutrition supports, peritoneal fluid, mesenteric lymph nodes （MLN）, liver, and pancreas were collected for bacterial culture with standard techniques to observe the incidence of bacterial translocation. Pathology changes of pancreas were analyzed by histopathologic grading and scoring of the severity of pancreas, and the degree of intestinal mucosal damage was assessed by measuring mucosal thickness, villus height, and crypt depth of ileum. Results Compared with PN and EEN, EIN significantly decreased the levels of serum amylase, serum aminotransferase, plasma endotoxin, and the incidence of bacterial translocation. Furthermore, compared with the others, the histology scores of inflammation in pancreas and the ileum injury （ileum mocosa thickness, villus height, and crypt depth） were significantly alleviated by EIN （P〈0.05）. Moreover, concerning liver function, the serum levels of alanine aminotransferase, aspartate aminotransferase and albumin were ameliorating significantly in the EIN group. Conclusion Our results suggested that EIN could maintain the integrity of intestinal mucosal barrier and reducing the incidence of bacterial translocation in SAP dogs. Early EIN was safe and more effective treatment for SAP dogs.

Effects of different doses of ginger-partitioned moxibustion on trefoil factor 1,mucin 5AC and epidermal growth factor receptor in rats with spleen deficiency syndrome

To observe the effects of different doses of ginger-partitioned moxibustion on serum trefoil factor 1 （TFF1） and mucin 5AC （MUCSAC） levels, as well as the expression of epidermal growth factor receptor （EGFR） in gastric mucosa of rats with spleen deficiency syndrome, therefore, to explore the possible mechanism and the dose-effect characteristics of ginger-partitioned moxibustion in spleen deficiency syndrome. Methods： Seventy-five SPF grade Sprague-Dawley （SO） rats were randomly divided into a blank control group （group A）, a model group （group B）, a 3 moxa-cone ginger-partitioned moxibustion group （group C1）, a 6 moxa-cone ginger-partitioned moxibustion group （group C2） and a 9 moxa-cone ginger-partitioned moxibustion group （group C3） using random number table method, 15 rats in each group. Except group A, rats in the other groups received intragastric administration of 4 ~C 200% concentrated Da Huang （Radix et RhizomaRhei） to prepare spleen deficiency syndrome model. After successful modeling, rats in group B received no treatment; rats in group C1, C2 and C3 were treated with 3, 6 and 9 moxa-cone ginger-partitioned moxibustion at Zusanli （ST 36）and Zhongwan （CV 12） respectively for 8 continuous days. The general symptom score of rats was observed. The serum levels of TFF1 and MUC5AC were detected by enzyme-linked immunosorbent assay （ELISA）. The expression of EGFR protein in gastric mucosa was detected by immunohistochemistry. Results： After the treatment, compared with group A, the spleen deficiency symptom score was increased in group B, the levels of serum TFF1 and MUC5AC, the EGFR protein expression in gastric tissues of group C1, C2 and C3 were significantly increased （all P〈0.01）; compared with group B, the spleen deficiency scores were decreased in group C1, C2 and C3, and the serum levels of TFF1 and MUC5AC, as well as EGFR protein expression in gastric tissues were increased （all P〈0.01）. Compared with group C1, the spleen deficiency scores were decreased in group C2 and C3, the serum levels of TFF1 and MUC5AC, and the expression of EGFR protein in gastric tissues were increased （all P〈0.01）, however, there was no significant difference between group C2 and C3 （all P〉0.05）. The mechanism may be related to the increase of serum TFF2 and MUC5AC levels and activation of EGFR protein. Conclusion： Ginger-partitioned moxibustion can improve the symptoms, as well as promote the proliferation and repair of gastric mucosa in rats with spleen deficiency. The therapeutic efficacy of 6 or 9 moxa-cone ginger-partitioned moxibustion is better than that of 3 moxa-cone ginger-partitioned moxibustion, while the efficacies are equivalent between 6 and 9 moxa-cone Ringer-Dartitioned moxibustion groups.

Effect of moxibustion stimulation on repair of injured gastric mucosa after common peroneal nerve transection

Objective： To investigate the protective effect of moxibustion in initiating the endogenous protection information on gastric mucosa, and its relationship with the pathway of common peroneal nerve. Methods： Forty-eight Sprague-Dawley（SD） rats were randomly divided into a normal group（group A）, a model group（group B）, a moxibustion model group（group C） and a moxibustion model plus surgery group（group D）, 12 in each group. Except for group A, rats in the other groups were treated with dehydrated ethanol and aspirin to prepare gastric mucosal damage model. The rats in group B were not treated with any interventions; rats in group C received moxibustion at Zusanli（ST 36）, twice a day for continuous 3 d. The rats in group D were subjected to preparing the gastric mucosal damage model after the common peroneal nerve transection, followed by moxibustion at Zusanli（ST 36）. After a 3-day intervention, ulcer index（UI） in each group was observed, and the levels of gastric mucosa-related repair cytokines of tumor necrosis factor-α（TNF-α）, interleukin-4（IL-4） and heat shock protein 70（HSP70） were detected. Results： Compared with group A, the pathological changes and UI of group B were worse（P=0.000）, but TNF-α in serum and tissue was changed significantly（P=0.000, P=0.002）, IL-4 in serum and tissue was improved significantly（P=0.000, P=0.000）. Compared with group B, TNF-α and IL-4 in group C and group D were significantly improved（TNF-α： P=0.003, P=0.016; IL-4： P=0.000, P=0.002）. Compared with group C, the changes of UI in group B and group D were poor（both P=0.000）; the levels of TNF-α and IL-4 in serum were significantly decreased（TNF-α： P=0.000, P=0.025; IL-4： P=0.000, P=0.034）; and tissue HSP70 levels were decreased significantly（P=0.000, P=0.033）. Conclusion： Zusanli（ST 36） can transmit information through the pathway of common peroneal nerve, regulate the release of gastric mucosal protective factors, and up-regulate the expression of cytothesis-related proteins, so as to achieve the effect in repairing gastric mucosa.

Effect of herbal cake-partitioned moxibustion on MEK1/2 and ERK1/2 expressions of gastric tissues in rats with spleen deficiency syndrome

Objective： To observe the effect of herbal cake-partitioned moxibustion on the expressions of mitogen-activated protein kinase （MEK：1/2） and extracellular regulatory protein kinase （ERK：1/2） in gastric tissues of rats with spleen deficiency syndrome, and to explore the possible mechanisms of herbal cake-partitioned moxibustion in treating spleen deficiency syndrome. Methods： Sixty Sprague-Dawley （SD） rats were randomly divided into a blank control group （group A）, a model group （group B）, a ranitidine group （group C）, and a herbal cake-partitioned moxibustion group （group D） by random digit, 15 rats in each group. Rat models of spleen deficiency syndrome were made by intragastric administration of 4℃ 200% concentrated Da Huang （Radix et Rhizoma Rhei）. After successful modeling, the rats in group C were treated with 25 mg/（kg.bw） ranitidine by intragastric adminstration and rats in group D were treated with herbal cake-partitioned moxibustion at Zusanli （ST 36） and Zhongwan （CV 12）, for 8 d. Excepted for rats in group A, all the other rats were treated with indomethacin at 5 mg/（kg.bw） at 8：00 a.m. on the second day after finishing all the intervention and sacrificed 7 h later to isolate the stomach. Histopathological changes of the gastric tissues were observed under light microscope after hematoxylin-eosin （HE） staining. The protein expressions of MEK：1/2 and ERK：1/2 in the gastric tissues were detected by immunohistochemistry. Results： After intervention, the gastric mucosal injury in group B was significantly severer than that in group A, with large breakage and ablating; the damage of gastric mucosa was decreased in group C compared with group B; the gastric mucosal surface remained relatively complete, and the status of breakage and ablating was significantly improved. After intervention, compared with group A, the protein expressions of MEK：1/2 and ERK：12 in gastric tissues of the other groups were significantly higher （P〈0.01）. Compared with group B, the protein expressions of MEK：1/2 and ERK：1/2 in group C and D were significantly higher （all P〈0.01）. Compared with group C, the protein expressions of MEK：1/2 and ERK：1/2 in group D were significantly higher （P〈0.01）. Conclusion： Herbal cake-partitioned moxibustion promotes the repair of gastric mucosa in rats with spleen deficiency syndrome, via improving protein expressions of MEK：1/2 and ERK：1/2 in gastric tissues, as well as activating MEK/ERK signaling pathway.

Effect of oxytocin on gastric ischemia-reperfusion injury in rats

The effect of peripherally administered oxytocin(OT)on gastric ischemia-reperfusion injury(GI-RI)and its possible mechanism were investigated.The Sprague-Dawley(SD)rats were randomly divided into different treatment groups(n=6).The animal GI-RI model was established by clamping the celiac artery for 30 min to induce ischemia and then released to allow reperfusion for 1 h,and the degree of GI-RI was assessed by scoring the gastric mucosal damage index(GMDI),the gastric fluid output,gastric fluid output,gastric acidity were measured and the surgical preparations of vagotomy and sympathectomy were used to investigate the possible mechanism of OT on GI-RI.The results were as follows.Compared with the control group(NS plus GI-R only,GMDI 121.33P10.40,n=6),the intra peritoneal(ip)administration of oxytocin(20,100μg/0.5 mL)obviously attenuated GI-RI(P<0.05),GMDI were 82.33P14.26,53.5P5.58 respectively(n=6);the gastric fluid output and the gastric acidity(evaluated by pH)of the control group were(430.17P87.36)μL,1.55P0.25(n=6),and those of the OT group were(102.45P48.00)μL,2.65P0.40(n=6)res pectively;differences had statistical significance(P<0.01).The effect of oxytocin was reversed by atosiban,a selective oxytocin receptor antagonist.The GMDI of the group given atosiban 10 min before OT was 138.17P24.06(n=6),which had no significant difference with the control group.Oxytocin further attenuated GI-RI after vagotomy and sympathectomy(GMDI 6.83P8.89,29.67P5.54,n=6),compared with the GI-R group and the oxytocin group(P<0.01).These results indicated that the oxytocin could significantly protect gastric mucosal against injury induced by ischemia-reperfusion,and the oxytocin receptor was involved.This effect of oxytocin may be mediated through the vagus and sympathetic nerve,and then lead to the reduction of gastric juice output and the depression of gastric acidity.