Extracts from Radix Astragali and Radix Rehmanniae promote keratinocyte proliferation by regulating expression of growth factor receptors.

Chinese herbal medicine has long been used as a treatment for wounds. However, the underlying cellular and molecular mechanisms remain largely unknown. In this study it was shown that the proliferation of keratinocytes, which is known to play an important role in wound healing as the major cell type in the epidermis, was promoted by three herbal extracts/natural compounds: NF3 (an extract from the mixture of Radix Astragali (RA) and Radix Rehmanniae (RR) in the ratio of 2:1), stachyose (an isolated compound from Radix Rehmanniae) and extract P2-2 (a sub-fraction from the extract of Radix Astragali). The effect of the herbal extracts/natural compounds on the growth of keratinocytes was not influenced by a high glucose level, a condition similar to diabetic patients who usually suffer from diabetic foot ulcers. Real time RT-PCR results showed that the expression of epidermal growth factor (EGF) receptor, but not transforming growth factor-ß (TGF-ß) receptor, was up-regulated by NF3. Moreover, treatments with the EGF receptor kinase inhibitor AG1478 and the MEK inhibitor U0126 resulted in the diminishment of the effect of the three herbal extracts/natural compounds on keratinocyte proliferation, indicating that EGF receptor might have a significant role in this action. This study has further elucidated the molecular mechanism under which herbal extracts/natural compounds exert their effects on the wound healing process.

Effects of Radix Astragali and Radix Rehmanniae, the components of an anti-diabetic foot ulcer herbal formula, on metabolism of model CYP1A2, CYP2C9, CYP2D6, CYP2E1 and CYP3A4 probe substrates in pooled human liver microsomes and specific CYP isoforms.

The present study investigated the effects of Radix Astragali (RA) and Radix Rehmanniae (RR), the major components of an anti-diabetic foot ulcer herbal formula (NF3), on the metabolism of model probe substrates of human CYP isoforms, CYP1A2, CYP2C9, CYP2D6, CYP2E1 and CYP3A4, which are important in the metabolism of a variety of xenobiotics. The effects of RA or RR on human CYP1A2 (phenacetin O-deethylase), CYP2C9 (tolbutamide 4-hydroxylase), CYP2D6 (dextromethorphan O-demethylase), CYP2E1 (chlorzoxazone 6-hydroxylase) and CYP3A4 (testosterone 6ß-hydroxylase) activities were investigated using pooled human liver microsomes. NF3 competitively inhibited activities of CYP2C9 (IC(50)=0.98mg/ml) and CYP3A4 (IC(50)=0.76mg/ml), with K(i) of 0.67 and 1.0mg/ml, respectively. With specific human CYP2C9 and CYP3A4 isoforms, NF3 competitively inhibited activities of CYP2C9 (IC(50)=0.86mg/ml) and CYP3A4 (IC(50)=0.88mg/ml), with K(i) of 0.57 and 1.6mg/ml, respectively. Studies on RA or RR individually showed that RR was more important in the metabolic interaction with the model CYP probe substrates. RR dose-dependently inhibited the testosterone 6ß-hydroxylation (K(i)=0.33mg/ml) while RA showed only minimal metabolic interaction potential with the model CYP probe substrates studied. This study showed that RR and the NF3 formula are metabolized mainly by CYP2C9 and/or CYP3A4, but weakly by CYP1A2, CYP2D6 and CYP2E1. The relatively high K(i) values of NF3 (for CYP2C9 and CYP3A4 metabolism) and RR (for CYP3A4 metabolism) would suggest a low potential for NF3 to cause herb-drug interaction involving these CYP isoforms.

Anti-inflammatory activities of a kolaviron-inhibition of nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha production in activated macrophage-like cell line.

Kolaviron (KV), a biflavonoid fraction from the seeds of Garcinia kola has been shown to posess antiinflammatory properties in animal models of inflammation. In this study, the effect of KV on carrageenan-induced paw edema was investigated in mice. Furthermore, the effects of KV on the production of the pro-inflammatory mediators- nitric oxide (NO) and tumor necrosis factor alpha (TNF-alpha) were examined in an activated macrophage-like cell lines, RAW 264.7 cells. Administration of KV prior to injection of carrageenan significantly reduced the paw inflammation in a dose-dependent manner. KV consistently inhibited in-vitro production of NO and secretion of TNF-alpha in a dose-dependent manner. In addition, KV reduced the production of PGE2 in LPS-stimulated macrophages. Viability of cells at all concentrations studied was unaffected as determined MTT [3-(4,5- dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide] cytotoxicity assay. These results suggest that KV has inhibitory effects on LPS-induced TNF-alpha, NO and PGE2 production.

Inhibition of acute experimental colitis by a crude extract and diets containing seeds of Garcinia kola (heckel).

The protective effect of Garcinia kola (GK) crude extract on acetic acid induced colitis in rats was investigated. Colitis, a form of inflammatory bowel disease (IBD) is characterized by inflammation of colon. The pathology in colitis includes disruption of crypt architecture, inflammation of crypts, frank crypt abscesses, and hemorrhage or inflammatory cells in the lamina propria. Since oxidative stress plays an important role in the etiology of Inflammatory Bowel diseases,and Garcinia kola (GK), have been shown to reduce oxidative stress in the rat stomach.The present study was designed to investigate the effects of Garcinia kola on ulcerative colitis (UC) induced by acetic acid. Albino rats were divided into five groups; Group one served as control, group two received Normal saline, group three received 2.5% ethanol while groups four and five were given 20mg/kg and 100mg/kg of crude extract of Garcinia kolarespectively. In another experiment, rats were fed for 2 weeks on normal rat diets but specially composed to contain 12.5%, 25% and 50% by weight of G kola. Colitis was induced by intra-rectal administration of 6% acetic acid. The colonic damage was elucidated by macroscopic damage scores; colon wet weight, stool consistency and colonic edema (thickness of the colon). Intra-luminal administration of 6% acetic acid resulted in observable clinical and macroscopic signs of colitis.Pre-orally administered of crude extract of GK significantly reduced the colonic damage score (p<0.001), colon weight (p<0.001), thickness of the colon (p<0.001) and diarrhea (p<0.001).Histological examinations also indicated a marked reduction in tissue injury and inhibition in neutrophil infiltration in rats pretreated with crude extract of Garcinia kola. Results of this investigation provide experimental evidence of Garcinia kola as an anti-colitis agent.

Chinese green tea ameliorates lung injury in cigarette smoke-exposed rats.

BACKGROUND: Epigallocatechin-3-gallate (EGCG), which has been shown to have potent antioxidant effect, comprises 80% of catechins in Chinese green tea. This study was to investigate whether cigarette smoke (CS) exposure would induce lung morphological changes and oxidative stress in the CS-exposed rat model, and whether Chinese green tea (Lung Chen tea with EGCG as its main active ingredient) consumption would alter oxidative stress in sera and lung leading to protection of CS-induced lung damage. METHODS: Sprague-Dawley rats were randomly divided into four groups, i.e. sham air (SA), 4% CS, 2% Lung Chen tea plus SA or 4% CS. Exposure to SA or 4% CS was performed for 1h/day for 56 days in ventilated smoking chambers. Sera and lung tissues were collected 24h after last CS exposure for histology and all biochemical assays. RESULTS: Airspace enlargement and goblet cell hyperplasia were observed after 56-day CS exposure alone, which were abolished in the presence of green tea consumption. Serum 8-isoprostane level was significantly elevated (p<0.01) as well as lung superoxide dismutase (SOD) and catalase activities in CS-exposed rats compared to SA-exposed rats (p<0.05), which returned to the levels of SA-exposed rats after Chinese green tea consumption. CONCLUSION: These results indicate that increased levels of systemic oxidative stress after CS exposure play an important role in the induction of lung damage. Chinese green tea may have the ability to suppress CS-induced oxidative stress that leads to protection of lung injury.

Protective effect of polysaccharides from Angelica sinensis on ulcerative colitis in rats.

Ulcerative colitis (UC) involves the dysregulation of intestinal mucosal immunity and imbalance between the reactive oxygen species (ROS) and the endogenous anti-oxidants. While the protective effects of Angelica sinensis (AS) polysaccharides on neutrophil-dependent gastric mucosal damage have been reported, similar protective effects on UC are still uncertain. Hence our study aimed to investigate the effects of AS polysaccharides on rats with acute UC induced by 2,4-dinitrobenzene sulphonic acid (DNBS) evaluated after 24 h. Intrarectal injection of DNBS significantly reduced the glutathione (GSH) content, increased malondialdehyde concentration and raised the amount of apoptotic cells in colon tissues, which were related to oxidative stress and attenuated by AS polysaccharides pretreatment (5 mg/ml and 10 mg/ml). These findings suggest that oxidative stress and GSH depletion are highly associated with the pathological mechanism of UC, and the protective effects of AS polysaccharides are closely related to the prevention of oxidative stress, which may occur during neutrophil infiltration in the pathological process of UC.

Regional hyperthermia of the abdomen in conjunction with chemotherapy for peritoneal carcinomatosis: evaluation of two annular-phased-array applicators.

BACKGROUND: Peritoneal carcinomatosis is a stage of gynecological and gastrointestinal malignancies with poor prognosis. Options for enhancing the effect of standard chemotherapy, such as aggressive surgery and intraperitoneal chemotherapy, have limitations. In this phase I/II study, we evaluated regional hyperthermia of the pelvis and abdomen using the annular-phased-array technique as an adjunct to chemotherapy. METHODS: Forty-five patients with peritoneal carcinomatosis (with or without liver metastases) in colorectal cancer (CRC) (n = 16), ovarian cancer (OC) (n = 17), or gastric/pancreatic/biliary cancer (n = 12) underwent standard chemotherapy and regional hyperthermia. Most CRC patients received second-line chemotherapy. All OC patients were platinum resistant. Regional hyperthermia was applied using a SIGMA-60 applicator (OC), a SIGMA-Eye/MR applicator (CRC), or various ring applicators (gastric/pancreatic/biliary cancer). RESULTS: Abdominal regional hyperthermia was well tolerated, with acceptable acute discomfort and no long-term morbidity. The SIGMA-Eye/MR applicator achieved higher systemic temperatures (associated with higher systemic stress) and more effective heating of the upper abdomen; the SIGMA-60 applicator achieved higher temperatures (and power densities) in the pelvis. Three-year overall survival was encouraging for patients with CRC (22%) and OC (29%) but not gastric/pancreatic/biliary cancer. For the SIGMA-60 applicator (patients with OC), higher measured temperatures at the vaginal stump correlated with better outcome. CONCLUSIONS. The SIGMA-60 and SIGMA-Eye/MR applicators are feasible for abdominal heating and have low toxicity. The SIGMA-60 applicator is specifically suitable for malignancies with high pelvic burden; the SIGMA-Eye/MR applicator better heats the upper abdomen, including the liver. Further randomized investigations are warranted.

Image artifacts during MRT hybrid hyperthermia--causes and elimination.

PURPOSE: Online MR-thermometry during hyperthermia can improve treatment control. It needs excellent image quality during hyperthermia treatment to get information from subtracted images. MATERIALS AND METHODS: For hybrid hyperthermia two high-frequency devices were used in combination working with different frequencies. The imaging was performed on a 1.5 T MR tomograph (Siemens Symphony, Quantum Gradienten, Maestro Class, Firma Siemens, Erlangen, Germany) at 64 MHz whereas hyperthermia was administered with a BSD 2000 3D unit utilizing a Sigma Eye applicator and a 12 channel DODECK transistor amplifier (BSD 2000, BSD-MC, Salt Lake City, Utah, USA) operating at 100 MHz. For analysing image artifacts a spectrum analyser (Hewlett Packard HP8591E) was used. RESULTS: Two different image artifacts, occurring during the use of this hybrid system, are described. The artifacts result from introduction of additional frequencies into the imager. Here we demonstrated the detection and elimination of these spurious frequencies in the context of two case studies. CONCLUSION: Hybrid hyperthermia requires excellent imaging for optimal operation. Additional frequencies causing image artifacts can be identified by use of a spectrum analyser. Once identified, these interfering frequencies can be eliminated with appropriate RF filters. With MRI quality control for hyperthermia systems with different treatment frequencies is possible.

Free radical-scavenging activity of Taiwanese native plants.

The 70% aqueous acetone extracts of ten Taiwanese native plants were evaluated by various antioxidant assays, including 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl (.OH) radicals, and reducing power assay. In the present study, extracts of Acer buerferianum var. formosanum, Cleyera japonica var. morii, Cyclobalanopsis stenophylla var. stenophylloides, and Machilus zuihoensis exhibited stronger activity against DPPH radicals, and their IC50 values ranged from 5.4 to 8.3 microg/ml. The ten selected extracts effectively inhibited the formation of .OH generated in the Fenton reaction system. Among the extracts whose reducing power activities were determined, A. buerferianum var. formosanum, C. japonica var. morii, C. stenophylla var. stenophylloides, Eriobotrya deflex, and M. zuihoensis showed high activity. The results indicate the 70% aqueous acetone extracts of A. buerferianum var. formosanum, C. japonica var. morii, C. stenophylla var. stenophylloides, and M. zuihoensis with great potency in these assay systems and may be candidates for the development of natural antioxidants.

Effect of polysaccharides from Angelica sinensis on gastric ulcer healing.

Our previous study showed that a crude extract from Angelica sinensis (ASCE), which mainly consisted of polysaccharides, significantly promoted migration and proliferation of normal gastric epithelial cells. These results strongly suggest that ASCE has a direct wound healing effect on gastric mucosa. However, there is no report concerning the effect of ASCE on gastric ulcer healing in animal models. In this study, we found that ASCE promoted ulcer healing. The area of the ulcer was reduced. This was accompanied with a significant increase in mucus synthesis when compared with the control. Angiogenesis was inhibited by the treatment of ASCE. Cell proliferation, ODC and EGFR protein expression was not affected in this process. Thus, the mechanism of how ASCE accelerates ulcer healing in addition to its effect on mucus synthesis remains to be investigated.

Protective role of heme oxygenase-1 on trinitrobenzene sulfonic acid-induced colitis in rats.

Preliminary studies showed that the inducible form of heme oxygenase (HO-1) was induced and played a protective role in the process of inflammation. The present study investigated the possible role of HO-1 in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in rats. We measured HO-1 activity in TNBS-induced colitis in rats and analyzed the severity of colitis along with altered HO activity by assessing lesion area and myeloperoxidase activity. HO-1 mRNA and protein expressions were determined at different time points after TNBS induction. Free radical production and inducible nitric oxide synthase (iNOS), which participate in oxidative injury, were also assayed. HO activity and HO-1 gene expression increased markedly after TNBS induction. Administration with tin mesoporphyrin (SnMP), a HO inhibitor, potentiated the colonic damage along with a reduction in HO-1 activity. Furthermore, the reduction of HO-1 expression by SnMP also enhanced reactive oxygen species and iNOS expression, both of which were dramatically increased after the TNBS enema. L-Arginine pretreatment further aggravated the injurious action of SnMP. Our results indicate that HO-1 plays a protective role in the colonic damage induced by the TNBS enema, and the preventive effects probably result from decreased free radical production and inhibition of iNOS expression in colonic tissues.

Protective effect of polysaccharides-enriched fraction from Angelica sinensis on hepatic injury.

A polysaccharides-enriched fraction from the root of Angelica sinensis, which is known for its antiulcer action on the gastrointestinal tract, was isolated and studied for its hepato-protective effect in rodents. Intra-gastric administration of Angelica sinensis polysaccharides-enriched fraction (AP) at the doses of 50 or 75 mg/kg dose-dependently prevented liver toxicity induced by acetaminophen in mice but did not affect the serum acetaminophen concentration. It normalized the rises of serum alanine transferase (ALT) and hepatic nitric oxide synthase (NOS) activities and the decrease of glutathione level in the liver. It also reduced the hepatic malondialdehyde (MDA) concentration. The protective effect was less evident in the carbon tetrachloride (CCl4)-treated animals including mice and rats. In the rat the elevated serum ALT level was unaffected though the MDA level was similarly reduced by the higher dose of AP. In these animals, CCl4 increased the hepatic glutathione level instead while the NOS activity remained unchanged. These findings suggest that the pathogenic mechanisms of both acetaminophen and CCl4 are different. AP is more effective in the protection against liver damage induced by acetaminophen, which is associated with the glutathione depletion and nitric oxide synthase activation in the liver.

A mechanistic study of proliferation induced by Angelica sinensis in a normal gastric epithelial cell line.

It has been reported that an extract from Angelica sinensis mainly consisting of polysaccharides (95%) prevented ethanol- or indomethacin-induced gastric mucosal damage (Cho CH et al. Planta Med 2000;66:348-51). However, it is not known whether Angelica sinensis has a direct stimulatory effect on the healing of gastric mucosal lesions. To study the hypothesis that Angelica sinensis has a direct mucosal healing effect in rats and in isolated gastric epithelial cells, we assessed the wound repair in both animals and normal cell culture (RGM-1), as well as [3H]thymidine incorporation, ornithine decarboxylase (ODC) activity, and ODC protein and c-Myc protein expression after different treatments in RGM-1 cells. We found that Angelica sinensis crude extract (ASCE) dose-dependently enhanced gastric ulcer healing in rats and promoted wound repair in RGM-1 cells. It also significantly stimulated [3H]thymidine incorporation and ODC activity in RGM-1 cells in a concentration-dependent manner. ODC and c-Myc protein expression was also increased as a result of this process. DL-alpha-difluoromethyl-ornithine repressed the [3H]thymidine incorporation and ODC activity induced by ASCE. Pretreatment with c-Myc antisense oligodeoxynucleotides blocked the stimulatory action of ASCE on [3H]thymidine incorporation and ODC protein expression. These data suggest that ASCE has a direct mucosal healing effect on gastric epithelial cells, while ODC and c-Myc are closely associated with this effect.

Aggravating effect of cigarette smoke exposure on experimental colitis is associated with leukotriene B(4) and reactive oxygen metabolites.

BACKGROUND/AIMS: Cigarette smoking is closely related to the development and recurrence of inflammatory bowel disease (IBD). The present study aimed to investigate the underlying mechanisms of the adverse action of cigarette smoke (CS) exposure on trinitrobenzene sulfonic acid (TNBS)-induced IBD. METHODS: Rats were preexposed to CS once daily for 4 days before receiving a TNBS enema, and they were killed 24 h afterwards. The colonic myeloperoxidase (MPO) and xanthine oxidase (XO) activities, leukotriene B(4) (LTB(4)) and glutathione (GSH) levels, as well as the production of reactive oxygen metabolites (ROMs) were measured. RESULTS: CS preexposure significantly augmented the adverse effects of the TNBS enema on colonic damage and increase in MPO activity, while it did not significantly alter the XO activity. Meanwhile, the elevation of ROM production and LTB(4) concentration in colonic tissues after the TNBS enema was also markedly enhanced by CS exposure. In contrast, the depressive action of the TNBS enema on cellular antioxidant GSH levels was reduced further by CS exposure. Pretreatment with a specific LTB(4) antagonist, ONO-4057, protected against colonic damage, particularly in the CS group. CONCLUSION: CS exposure aggravated experimental IBD. This adverse action could be due to the depletion of GSH together with overproduction of LTB(4), followed by the accumulation of neutrophils and ROMs in the colonic tissue.

Angelica sinensis modulates migration and proliferation of gastric epithelial cells.

A crude extract from Angelica sinensis (ASCE), which mainly consists of polysaccharides, prevents ethanol- or indomethacin-induced gastric mucosal damage and promotes ulcer healing. The aim of this study was to test the hypothesis that ASCE has a direct stimulating effect on gastric epithelial cells for wound healing. We found that ASCE significantly promoted the migration of epithelial cells over an artificial wound on the surface of an RGM-1 monolayer. The extract also stimulated DNA synthesis in a dose-dependent manner and concomitantly increased EGF mRNA expression. Co-incubation of ASCE with anti-EGF antibody reduced the speed of migration and the DNA synthesis, which however were still higher than the control without ASCE. These results strongly suggest that ASCE has a direct wound healing effect on gastric mucosa, and this is acting partially through an EGF-mediated pathway.

Beneficial intervention of experimental colitis by passive cigarette smoking through the modulation of cytokines in rats.

BACKGROUND: Epidemiologic observations have indicated that cigarette smoking decreases the risk of ulcerative colitis, but the modes of action remain anonymous. The present study aimed to investigate the beneficial effects of passive cigarette smoking using an animal colitis model. We hypothesized that the underlying mechanisms may involve immunoregulation of cytokines. METHODS: Experimental colitis was induced in rats by enema administration of 2,4-dinitrobenzene sulfonic acid (DNBS). Passive cigarette smoking by rats was performed for 1 hour once daily, from 3 days before DNBS enema until they were sacrificed on day 8. Other groups of DNBS-treated rats received therapeutic treatment of cyclosporin A or pentoxifylline, a tumor necrosis factor (TNF)-alpha inhibitor. Macroscopic and histologic damage were graded, and the colonic levels of different cytokines and the levels/activities of parameters related to neutrophil activation were also measured. RESULTS: DNBS-induced colonic damage was improved in passive-cigarette-smoking rats. This was accompanied by attenuation of the elevated colonic myeloperoxidase and inducible nitric oxide synthase activities and leukotriene B4 level. Likewise, the augmentation in colonic levels of TNF-alpha, interleukin (IL)-1 beta, and IL-6 in colitis rats was also alleviated by passive cigarette smoking. In contrast, the deprivation of colonic IL-10 during colitis was preserved in cigarette-smoking rats. These effects were similarly accomplished by pentoxifylline and, to some degree, by cyclosporin A. CONCLUSIONS: The results support the idea that the beneficial effects of passive cigarette smoking in experimental colitis involved immunoregulation of cytokines in colonic tissues.

Study of the gastrointestinal protective effects of polysaccharides from Angelica sinensis in rats.

We studied the protective effects of polysaccharides isolated from the root of Angelica sinensis (Oliv.) (Danggui) on gastrointestinal damage induced by ethanol or indomethacin in rats. Oral administration of ethanol provoked a marked hemorrhagic damage in the glandular mucosa, which was accompanied with a significant increase of myeloperoxidase (MPO) activity, a marker enzyme for inflammation and neutrophil infiltration. An extract from Angelica, which mainly consisted of polysaccharides (95%) (AP), dose-dependently prevented gastric mucosal damage. This ulcer protective effect could last at least 12 h after administration. Prostaglandin E2 produced a similar anti-lesion effect. AP and prostaglandin E2 also reduced mucosal MPO activity. Indomethacin-induced gastrointestinal damage, another neutrophil-dependent lesion model in the gastrointestinal tract, was also prevented by AP pretreatment. The present findings suggest that polysaccharides from Angelica possess an anti-inflammatory action, perhaps through the inhibitory action on neutrophil infiltration in the gastrointestinal mucosa. AP could potentially be useful to prevent any neutrophil-dependent mucosal injury in the gastrointestinal tract.

Involvement of neutrophils and free radicals in the potentiating effects of passive cigarette smoking on inflammatory bowel disease in rats.

BACKGROUND & AIMS: Cigarette smoking is associated with inflammatory bowel diseases (IBDs), particularly Crohn's disease, in humans. The aim of this study was to examine whether passive cigarette smoking aggravates experimental IBD in rats and to clarify the underlying mechanisms. METHODS: Rats were exposed to cigarette smoke (CS) for 1 hour once daily for 4 days before induction of IBD by 2,4, 6-trinitrobenzene sulfonic acid (TNBS)-ethanol enema and were then killed at 2, 6, or 24 hours later. RESULTS: Preexposure to CS significantly potentiated colonic damage induced by TNBS. TNBS-ethanol enema caused a pronounced increase in colonic myeloperoxidase activity, leukotriene B(4) level, and also inducible nitric oxide synthase activity, its protein, and messenger RNA expression. These parameters were all significantly increased further by exposure to CS. In contrast, increased colonic superoxide dismutase activity after TNBS-ethanol enema was attenuated by CS exposure. The potentiating effects of CS exposure on TNBS-induced IBD were significantly alleviated after pretreatment with cyclosporin A (an immunosuppressant), N (G)-nitro-L-arginine methylester (a nitric oxide synthase inhibitor), and dimethyl sulfoxide (a hydroxyl radical scavenger). CONCLUSIONS: The results show that promotion of neutrophil infiltration and free radical production contributed significantly to the potentiating effect of passive cigarette smoking on experimental IBD.

The correlation of the weakening effect on gastric mucosal integrity by 5-HT with neutrophil activation.

The effects of 5-hydroxytryptamine (5-HT) on ethanol-induced gastric mucosal damage and on epithelial and vascular integrity were investigated. Male Sprague-Dawley rats were administered with 5-HT (5 or 10 mg/kg, IP) 30 min prior to the challenge with ethanol (40% v/v, 10 ml/kg, PO). 5-HT dose dependently aggravated ethanol-induced injury in the gastric mucosa. Both xanthine oxidase (XO) and myeloperoxidase (MPO) activities in the mucosa were significantly increased with the high dose of 5-HT, which also potentiated the elevation of these enzyme activities by ethanol. However, the mucosal superoxide dismutase activity was left unaltered. In neutropenic (antineutrophil serum-treated) animals, the ethanol-induced gastric mucosal injury was significantly ameliorated, with or without the pretreatment of 5-HT (10 mg/kg). In addition, the effect of 5-HT on the activity of MPO, but not of XO, was also attenuated in these animals. In the ex vivo gastric chamber study on pentobarbital-anesthetized animals, volume of gastric secretion was significantly decreased in the 5-HT-treated groups, with further reduction after ethanol incubation. Transmucosal potential difference (PD) was significantly reduced in 5-HT-treated rats, which also potentiated the ethanol-induced drop in PD. Nevertheless, 5-HT dose dependently increased mucosal vascular permeability and further enhanced during ethanol incubation. These findings suggest that 5-HT adversely affects the defense mechanisms of the gastric mucosa by reducing the secretory function of the mucosal cells and to weaken the epithelial and vascular integrity. Neutrophil activation appears to be responsible for the detrimental effects of 5-HT partly through the elevation in MPO activity. The increase in mucosal XO activity by 5-HT may induce free radical production and possibly modulate the ulcerogenic processes.

Mucosal nitric oxide is not responsible for the hemodynamic changes induced by nicotine in rat stomachs.

It has been shown that chronic nicotine treatment decreases gastric mucosal blood flow (GMBF). The mechanism for this action is still not defined. In this study, nicotine treatment (5, 25 or 50 µg/ml drinking water) for 10 days dose dependently reduced the GMBF and volume of hemoglobin but increased ethanol-induced gastric damage. These effects were potentiated by N(ω)-nitro-l-arginine methyl ester (l-NAME), a nitric oxide (NO) synthase inhibitor. l-arginine but not the d-analog restored the actions of l-NAME, indicating that the selective action of l-NAME. However, the potentiating actions of l-NAME were significantly attenuated in the nicotine-pretreated rats. When the basal mucosal NO synthase (both iNOS and cNOS) activity and its second messenger cyclic GMP levels were measured, no difference was found between the nicotine and the non-nicotine groups. Furthermore, high dose of l-arginine could not reverse the action of nicotine. These findings suggest that the adverse action of chronic nicotine treatment on GMBF and lesion formation is probably mediated through a NO independent mechanism.