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1.
Two aspirin tablets in 100 ml fluid will produce microscopical damage to the human stomach. A study was performed to determine whether a small amount of sodium bicarbonate (equivalent to one-third of a teaspoonful of baking soda) could protect against this damage. Sequential gastric biopsy specimens were taken from 15 healthy subjects before, during, and after intragastric instillation of one of the following isotonic solutions: saline; sodium bicarbonate; 600 mg aspirin suspended in sodium bicarbonate; and aspirin suspended in saline. On a separate day the same solutions were instilled, but gastric transmucosal potential differences were monitored. Light microscopy and scanning electron microscopy of the biopsy specimens showed occasional mucous degranulation of mucosal surface cells, but no cell damage during instillation of sodium bicarbonate. Light microscopy studies 10 minutes after aspirin in saline showed damage in 20% of surface cells, with focal areas of cellular disruption and microscopic erosions, but only 3·4% of cells were damaged after aspirin in bicarbonate and there were no erosions. Electron microscopy showed a damaged honeycombed appearance of surface epithelium after aspirin in saline and a normal cobblestone appearance after aspirin in bicarbonate. Aspirin dissolved in bicarbonate failed to induce the usual fall in potential difference.These findings indicate that sodium bicarbonate in amounts equivalent to one-third of a teaspoonful of baking soda protects the gastric mucosa against aspirin-induced damage and prevents the usual fall in potential difference after aspirin. 相似文献
2.
《Prostaglandins》1981
The relationship between gastro-intestinal damage and the inhibition of cyclo-oxygenase by non-steorid anti-inflammatory drugs is discussed. In anti-inflammatory doses, aspirin and the newer substitutes, indomethacin, ketoprofen, naproxen and flurbiprofen, which reduce prostaglandin levels in inflammatory exudates likewise inhibit cyclo-oxygenase activity in the gastric mucosa. These compounds all induce gastric damage. In contrast, sodium salicylate and a novel compound BW755C inhibit prostaglandin production in the inflamamtory exudate, yet fail to inhibit gastric cyclo-oxygenase in the mucosa and do not form gastric erosions. Studies on the duration of cyclo-oxygenase inhibition with indomethacin indicate a close correlation between the rate of healing of gastric erosions and the recovery of cyclo-oxygenase activity in the gastric mucosa. No such relationship is seen in the small-intestine. The hydroperoxy metabolites of arachidonic acid, formed by the lipoxygenase enzymes, have vasodilator actions in the gastric circulation and the possible roles of lipoxygenase products in the gastric mucosa and their relationship to gastro-intestinal damage are discussed. 相似文献
3.
Aspirin-induced gastric injury in the rat: histologic changes and sucralfate cytoprotection 总被引:12,自引:0,他引:12
P H Guth G Paulsen 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1987,184(4):423-428
The effect of pretreatment with intragastric sucralfate on aspirin acid-induced gastric mucosal lesions in the rat was studied. The finding by others that sucralfate is cytoprotective and that this cytoprotective effect probably is mediated, at least in part, by stimulation of endogenous prostaglandin synthesis was confirmed. In addition, a time course study revealed that the maximum cytoprotective effect was present 1 min after sucralfate administration and persisted for at least 6 hr. Microscopic evaluation of histologic sections revealed that sucralfate significantly decreased aspirin-induced deep mucosal erosions (those extending into the parietal cell area) but not superficial mucosal damage. Superficial mucosal damage (surface cell injury and erosions involving the mucous neck cell area) could not be detected grossly. The lesions seen grossly were deeper erosions involving the parietal cell area of the mucosa. 相似文献
4.
Our previous investigations suggest that the reduction of stress-induced corticosterone release, or inhibition of corticosterone actions, promotes stress-induced gastric erosions in rats. In this study the effect of glucocorticoid deficiency on susceptibility to gastric mucosal injury by nonsteroidal antiinflammatory drugs (NSAIDs) was evaluated in rats. Gastric erosions induced in male rats by indomethacin (25 mg/kg sc) or acidified aspirin (40 mM po) were studied one week after adrenalectomy with or without corticosterone replacement or after occupation of glucocorticoid receptors by the antagonist RU-38486 during the period of erosion formation. Corticosterone for replacement (4 mg/kg sc) was injected 15 min before the administration of indomethacin or acidified aspirin to adrenalectomized rats. The antagonist RU-38486 (10 mg/kg po) was administered twice, 20 min before and 60 min after NSAID administration. Plasma corticosterone levels were measured by fluorometry. Gastric erosions were quantitated by measuring the area of damage. Indomethacin or acidified aspirin induced both plasma corticosterone rise and gastric erosions. Adrenalectomy decreased both basal and NSAID-induced corticosterone levels and markedly promoted gastric erosion formation caused by the NSAID. An acute corticosterone replacement mimicking indomethacin-and aspirin-induced corticosterone rise prevented the effect of adrenalectomy on the gastric erosions. The administration of the glucocorticoid/progesterone antagonist RU-38486 significantly potentiated the formation of gastric erosions induced by indomethacin as well as aspirin. These observations suggest a gastroprotective action of glucocorticoids released in response to NSAID treatment against NSAID-induced injury. 相似文献
5.
A Bennett 《Acta physiologica Hungarica》1989,73(2-3):179-183
Much of the research on gastric mucosal protection has concerned prostaglandins. Some of the recent studies consolidate aspects first investigated a few years ago, but whose importance is now becoming established more clearly. This short review will mention some of the more recent work demonstrating the importance of prostaglandins in preventing stasis of gastric mucosal blood flow, effects on cell senescence and exfoliation, and the protection of a severe mucosal lesion by a mucus-containing plug which facilitates healing. The leukotrienes are other substances formed in the gastric mucosa from the same precursors as the prostaglandins. Their roles are not well understood, but may include participation in gastric inflammation, and in mucosal damage by nonsteroidal anti-inflammatory drugs (NSAIDs) and ethanol. The NSAIDs may damage the gastric mucosa not only by reducing the formation of protective prostaglandins, but also by increasing the metabolism of prostaglandin precursors into leukotrienes. Another factor is thromboxane A2, a substance that is damaging to the gastric mucosa but whose synthesis is inhibited by NSAIDs. The prostaglandin analogues produced for the treatment of peptic ulcer may find a major use in the protection against damage by NSAIDs. Not only may they act as 'replacement therapy' for the inhibited prostaglandins, but they protect against damage from substances that do not inhibit prostaglandin synthesis. In doses that raise the gastric pH, the prostaglandins reduce the local absorption of NSAIDs by increasing their ionisation. In rats, paracetamol protects against damage by aspirin, but whether this occurs in man is controversial. Work not previously published demonstrates that paracetamol does not affect the inhibition of prostaglandin formation by indomethacin in human isolated gastric mucosa. 相似文献
6.
H Tanimoto K Yoshida H Yokozaki W Yasui H Nakayama H Ito K Ohama E Tahara 《Virchows Archiv. B, Cell pathology including molecular pathology》1991,61(4):263-267
The expression of mRNA for the basic fibroblast growth factor (FGF) gene was examined in seven human gastric carcinoma cell lines and in tissue from 29 gastric carcinomas together with the adjacent normal mucosa. Among the seven gastric carcinoma cell lines, the MKN45 cell line expressed mRNA for the basic FGF gene. Basic FGF protein production was confirmed by flow cytometric analysis and immunohistochemistry. Among the surgical specimens, 16 (55%) of 29 gastric carcinomas showed higher levels of basic FGF mRNA than the normal mucosa. Interestingly, in scirrhous gastric carcinomas characterized by their fibrous stroma and rapid growth, 9 (69%) of 13, samples examined revealed higher levels of basic FGF mRNA than normal mucosa, whereas only 3 (33%) of the 9 well differentiated adenocarcinomas studied produced similar results. Immunohistochemically, basic FGF protein was localized in tumor cells. These results suggest that basic FGF produced by tumor cells may play an important role in producing fibrosis and angiogenesis in gastric carcinomas. 相似文献
7.
Alderman BM Cook GA Familari M Yeomans ND Giraud AS 《American journal of physiology. Gastrointestinal and liver physiology》2000,278(6):G839-G846
Adaptation of the gastric mucosa to nonsteroidal anti-inflammatory drug-induced injury is a well-documented phenomenon, but the mechanisms are not known. We investigated whether changes in stress protein expression and apoptosis play roles in adaptation of rat stomach to aspirin. RT-PCR and Western blotting techniques were used to analyze mRNA and protein expression of HSP72 and HSP90 and cleavage of caspase 3 protein. Apoptosis was detected by the TUNEL method and quantified. HSP72 mRNA and protein expression was unchanged in adapted mucosa, whereas HSP90 mRNA and protein levels decreased. Caspase 3 protein was activated, and the number of apoptotic cells increased in mucosa after one aspirin dose. However, in adapted mucosa after aspirin, activated caspase 3 and the number of apoptotic cells had returned to basal levels. Induction of the stress response was found not to be a mechanism of mucosal adaptation against multiple doses of aspirin. Our results lead us to propose instead that resistance to aspirin-induced apoptosis plays a role in the protective phenomenon of adaptation. 相似文献
8.
J I Paz-Bouza I García M M Abad E Mu?oz M R Rincón A Bullón 《Histology and histopathology》1991,6(4):485-489
Biopsy specimens of gastric and duodenal mucosa from 326 patients were examined bacteriologically and histologically to determine the correlation between chronic gastritis and H. pylori colonization. H. pylori was identified in 111 (66.5%) patients with evidence of chronic gastritis and in 97 (82.2%) individuals who had gastritis associated with other pathology (gastric o duodenal ulcer, carcinoma o bulboduodenitis). The spiral bacteria was found more frequently in specimens with chronic superficial gastritis (88/107) and no significant difference was observed between the grade of activity of gastritis and H. pylori colonization. Giemsa stain was the most suitable method for detecting H. pylori in histological sections. By electron microscopy the microorganism was seen on the surface of the gastric mucosa, beneath the mucous layer, and more occasionally in intercellular junctions and the gastric pit. 相似文献
9.
Hirotoshi Tanimoto Kazuhiro Yoshida Hiroshi Yokozaki Wataru Yasui Hirofumi Nakayama Hisao Ito Koso Ohama Eiichi Tahara 《Virchows Archiv. B, Cell pathology including molecular pathology》1992,61(1):263-267
The expression of mRNA for the basic fibroblast growth factor (FGF) gene was examined in seven human gastric carcinoma cell
lines and in tissue from 29 gastric carcinomas together with the adjacent normal mucosa. Among the seven gastric carcinoma
cell lines, the MKN45 cell line expressed mRNA for the basic FGF gene. Basic FGF protein production was confirmed by flow
cytometric analysis and immunohistochemistry. Among the surgical specimens, 16 (55%) of 29 gastric carcinomas showed higher
levels of basic FGF mRNA than the normal mucosa. Interestingly, in scirr-hous gastric carcinomas characterized by their fibrous
stroma and rapid growth, 9 (69%) of 13, samples examined revealed higher levels of basic FGF mRNA than normal mucosa, whereas
only 3 (33%) of the 9 well differentiated adenocarcinomas studied produced similar results. Immunohistochemically, basic FGF
protein was localized in tumor cells. These results suggest that basic FGF produced by tumor cells may play an important role
in producing fibrosis and angiogenesis in gastric carcinomas. 相似文献
10.
Jie Du Xiao-Hui LiWang Zhang Yong-Mei YangYue-Han Wu Wen-Qun LiJun Peng Yuan-Jian Li 《Biochemical and biophysical research communications》2014
Large-dose or long-term use of aspirin tends to cause gastric mucosa injury, which is recognized as the major side effect of aspirin. It has been demonstrated that glutamate exerts a protective effect on stomach, and the level of glutamate is critically controlled by cystine/glutamate transporter (Xc−). In the present study, we investigated the role of glutamate–cystine/glutamate transporter system in aspirin-induced acute gastric mucosa injury in vitro and in vivo. Results showed that in human gastric epithelial cells, aspirin incubation increased the activity of LDH and the number of apoptotic cells, meanwhile down-regulated the mRNA expression of Xc− accompanied with decreased glutamate release. Similar results were seen in a rat model. In addition, exogenous l-glutamate attenuated the gastric mucosa injury and cell damage induced by aspirin both in vitro and in vivo. Taken together, our results demonstrated that acute gastric mucosa injury induced by aspirin is related to reduction of glutamate–cystine/glutamate transporter system activity. 相似文献
11.
G. Faulkner P. Prichard K. Somerville M. J. Langman 《BMJ (Clinical research ed.)》1988,297(6659):1311-1313
A case-control study was performed to determine whether aspirin confers a similar risk of bleeding from gastric or duodenal ulcers in the elderly as non-aspirin, non-steroidal anti-inflammatory drugs. The intake of analgesics in 230 patients with bleeding ulcers aged 60 and over and in hospital and community controls matched for age and sex was examined. Those who had taken aspirin were between two and three times more likely to be admitted to hospital with bleeding ulcers. This increased risk was not accounted for by aspirin taken for indigestion or by concurrent use of non-aspirin, non-steroidal anti-inflammatory drugs. A similar effect was not seen for paracetamol. When aspirin and other non-steroidal anti-inflammatory drugs were considered together the overall risk attributed to the drugs suggested that these drugs may be responsible for over a third of admissions for bleeding peptic ulcers in the elderly. 相似文献
12.
H Goto S Sugiyama Y Kawabe M Kuroiwa A Ohara Y Tsukamoto S Nakazawa T Ozawa 《Biochemistry international》1990,20(6):1119-1125
This study was designed to analyze PGs in human gastric mucosa using biopsy specimens at femtomole level by the combination of microcolumn HPLC and He/Cd laser induced fluorescence detection. Biopsy specimens were taken along the greater curvature at the corpus of the stomach, in which no gastric disease was revealed by endoscopic examination. PGs extracted from human gastric mucosa were derivatized with ADAM, and ADAM-derivatized PGs were injected into the column for analysis. The mobile phase of acetonitrile-water (73:27) containing 0.01% of phosphoric acid was used at a constant pressure of 20 kgf/cm2. Using this system, PGs in few mg of human gastric mucosa obtained by biopsy were well separated and detected; i.e., 1653 +/- 254 (femtomole/mg tissue), 279 +/- 56, 729 +/- 153, 831 +/- 199 for 6-keto-PGF1 alpha, PGF2 alpha, PGE2, and PGD2, respectively. In conclusion, the microcolumn HPLC system with laser induced fluorescence detection is a reliable method for determining individual PGs in human gastric mucosa. In addition, PGI2 is the predominant PG in human gastric mucosa and probably plays an important role in gastric function. 相似文献
13.
《Life sciences》1997,62(1):PL13-PL18
The influence of pretreatment with (R)-α-methylhistamine, selective agonist of histamine H3 receptors, has been investigated on gastric mucosal lesions at different time intervals, from 5 to 60 minutes, after administration of absolute ethanol in the rat. The amount and depth of lesions were quantitatively evaluated by light microscopy. In rats pretreated with (R)-α-methylhistamine, the integrity of the mucosa was preserved in approximately 80% of the total mucosal length measured despite ethanol challenge. Prevention of lesion formation was as great at 5 min after ethanol administration as at 60 min. When present, damage involved mainly superficial mucosa and lesions extending deeply into the gland region were evident in 1–2.5% of the total mucosa. Present findings indicate that mechanisms by which (R)-α-methylhistamine operates enable the mucosa to counteract damage just from the moment of exposure to ethanol and that protection is exerted both on surface and pit cells and on gastric glands. 相似文献
14.
M Danova G Mazzini G Wilson G Ucci P Dionigi A Riccardi R Fiocca 《Basic and applied histochemistry》1987,31(1):73-82
Flow cytometric DNA analysis was performed on both fresh and on paraffin embedded samples obtained by gastroscopic biopsies in 5 patients with histologically normal gastric mucosa (20 specimens) and by radical gastrectomies in 9 cases of human gastric cancer (36 specimens). Ploidy and the distribution of cells in the different cell cycle phases were estimated. Results from fresh tissue were compared with those from paraffin embedded material. All the samples of normal mucosa showed a diploid modal DNA content. Unimodal DNA distribution was found both in fresh and paraffin embedded material of 3 well differentiated gastric adenocarcinomas, whereas the remaining 6 tumors (1 moderately differentiated, 4 poorly differentiated and 1 undifferentiated adenocarcinomas) showed aneuploidy. The percentage of cells in S phase in normal tissue and in tumors were respectively 9.0% and 11.9% in the fresh material, and 10.0% and 15.0% in the paraffin embedded material. No statistically significant differences were found between fresh and paraffin embedded samples, whereas the proliferative activity was, in both cases, statistically higher in tumors than in normal mucosa (p less than .01 and p less than .001 respectively). The quality of DNA flow cytometry from paraffin embedded material was comparable with that from fresh samples. 相似文献
15.
Neurogenic dystrophy of the gastric mucosa was induced by a 3-hour immobilization and electrostimulation. By the end of the stimulation there developed in the gastric mucosa hemorrhagic erosions which persisted for two days after the stimulation. According to the data of macro- and microscopic studies injections of 1-DOPA to rats in a dose of 10 mg/kg of body weight for a period of two days after the cessation of stimulation accelerated the healing of hemorrhagic erosions of the gastric mucosa. 相似文献
16.
Immunohistochemical Detection of Helicobacter pylori in the Surface Mucous Gel Layer and Its Clinicopathological Significance 总被引:2,自引:0,他引:2
Background Attempts have been made to develop an accurate method for detecting Helicobacter pylori in histological sections.
Materials and Methods. Biopsy specimens were obtained from the stomachs of 167 patients with gastric ulcer (33), duodenal ulcer (52), gastroduodenal ulcer (15), chronic gastritis (45), and normal mucosa (22) before antimicrobial treatment and from 108 of these patients after treatment. Biopsy specimens were (1) cultured, (2) fixed in 10% buffered formalin, or (3) fixed in Carnoy's solution. The latter method was employed to preserve the surface mucous gel layer (SMGL) covering gastric surface mucous cells. Histological sections were stained with hematoxylin and eosin (H&E), with immunostaining using a commercially available polyclonal anti- H. pylori antibody.
Results. Cultures were positive for H. pylori in 61% of the cases before treatment and in 16% after treatment; by H&E staining using formalin-fixed materials: 70% and 9%; by immunostaining using formalin-fixed materials: 78% and 21%; and by immunostaining using Carnoy-fixed materials: 85% and 41% of biopsy speciemens, respectively. The difference in detection rates between materials fixed in formalin and those in Carnoy's solution was due to the detection of H. pylori in the SMGL by the latter, especially after antimicrobial treatment.
Conclusions. Immunostaining for H. pylori using materials fixed in Carnoy's solution revealed H. pylori in the SMGL as well as on the surface mucous cells and in the gastric pits and permitted the optimal detection of H. pylori in tissue sections. 相似文献
Materials and Methods. Biopsy specimens were obtained from the stomachs of 167 patients with gastric ulcer (33), duodenal ulcer (52), gastroduodenal ulcer (15), chronic gastritis (45), and normal mucosa (22) before antimicrobial treatment and from 108 of these patients after treatment. Biopsy specimens were (1) cultured, (2) fixed in 10% buffered formalin, or (3) fixed in Carnoy's solution. The latter method was employed to preserve the surface mucous gel layer (SMGL) covering gastric surface mucous cells. Histological sections were stained with hematoxylin and eosin (H&E), with immunostaining using a commercially available polyclonal anti- H. pylori antibody.
Results. Cultures were positive for H. pylori in 61% of the cases before treatment and in 16% after treatment; by H&E staining using formalin-fixed materials: 70% and 9%; by immunostaining using formalin-fixed materials: 78% and 21%; and by immunostaining using Carnoy-fixed materials: 85% and 41% of biopsy speciemens, respectively. The difference in detection rates between materials fixed in formalin and those in Carnoy's solution was due to the detection of H. pylori in the SMGL by the latter, especially after antimicrobial treatment.
Conclusions. Immunostaining for H. pylori using materials fixed in Carnoy's solution revealed H. pylori in the SMGL as well as on the surface mucous cells and in the gastric pits and permitted the optimal detection of H. pylori in tissue sections. 相似文献
17.
As to earlier observations that beta-carotene prevents the development of gastric mucosal injury produced by different noxious agent, however, its cytoprotective effect can be abolished by acute surgical vagotomy. The aim of this study was to evaluate the possible correlation between the gastric mucosal cytoprotective effect of beta-carotene and its gastric mucosal level in rats treated with IND. The gastric mucosal damage was produced by the administration of IND (20 mg/kg s.c.). The instillation of beta-carotene and acute surgical vagotomy (ASV) or SHAM operation were carried out 30 min before IND treatment. The rats were sacrificed 4 h after IND application, and the number and severity of gastric mucosal erosions were noted. The blood rats was collected quantitatively, the liver and the gastric mucosa were removed, and the beta-carotene and vitamin A level of the gastric mucosa, serum and liver were measured with HPLC. It was found that: 1. Beta-carotene induced gastric cytoprotection in SHAM-operated rats treated with IND but its effect disappeared after ASV. 2. Although the beta-carotene level of the gastric mucosa increased its concentration was not elevated in the serum of intact and vagotomized animals either. 3. Vitamin A Formation was not detected in the liver of animals with or without ASV. It was concluded that the lack of intake of beta-carotene into the gastric mucosa can not play etiologic role in the failure of gastric cytoprotection of rats with acute bilateral surgical vagotomy. 相似文献
18.
Tétreault MP Chailler P Beaulieu JF Rivard N Ménard D 《Journal of cellular physiology》2008,214(2):545-557
Restitution is a crucial event during the healing of superficial injury of the gastric mucosa involving epithelial cell sheet movement into the damaged area. We demonstrated that growth factors promote the restitution of human gastric epithelial cells. However, the intracellular signaling pathways that transmit extracellular cues as well as regulate basal and growth factor-stimulated gastric epithelial cell migration are still unclear. Herein, confluent human gastric epithelial cell monolayers (HGE-17) or primary cultures of gastric epithelial cells were wounded with a razor blade and the migration response was analyzed in presence or absence of TGFalpha or of pharmacological inhibitors of signaling proteins. Kinase activation profile analysis and phase-contrast microscopy were also performed in parallel. We report that ERK1/2 and Akt activities are rapidly stimulated following wounding of HGE-17 cells. Treatment of confluent HGE-17 cells or primary cultures of gastric epithelial cells with the phosphatidylinositol 3-kinase inhibitor LY294002, but not the MEK1 inhibitor, PD98059, significantly inhibits basal and TGFalpha-induced migration following wounding. Conversely, treatment of wounded HGE-17 cells with phosphatidylinositol(3,4,5)-triphosphate is sufficient to stimulate basal cell migration by 235%. In addition, pp60c-src kinase activity and tyrosine phosphorylation of epidermal growth factor receptors (EGFR) are also rapidly enhanced after wounding and pharmacological inhibition of both these activities strongly attenuates basal and TGFalpha-induced migration as well as Akt phosphorylation levels. In conclusion, the present results indicate that EGFR-dependent PI3K activation promotes restitution of wounded human gastric epithelial monolayers. 相似文献
19.
Impaired generation of prostaglandins from isolated gastric surface epithelial cells in portal hypertensive rats 总被引:2,自引:0,他引:2
We studied generation of prostaglandins E2 and 6-keto F1a by surface epithelial cell isolated from the gastric mucosa of portal hypertensive and sham-operated rats. Oxygenated cell suspensions containing 80 +/- 3% of surface epithelial cells were incubated for 30 min at 37 degrees C and the concentration of prostaglandin E2 and 6-keto-prostaglandin F1a in medium was measured by radioimmunoassay. Viability of the cells was assessed with Fast green exclusion at baseline and after 30-min and 60-min incubation. Within 30 minutes the surface epithelial cells obtained from portal hypertensive rats generated 22.0 +/- 1.6 (mean +/- SE) pg prostaglandin E2 and 40.7 +/- 4.7 pg 6-keto prostaglandin F1a, per 10(6) cells. These were significantly less than prostaglandin generation by cells obtained from sham-operated rats. The viability of the surface epithelial cells from portal hypertensive rats was also significantly reduced compared with sham-operated rats after 60 minute incubation. Reduced ability of the surface epithelial cells to generate prostaglandins may be one mechanism for increased susceptibility of portal hypertensive gastric mucosa to injury by noxious agents. 相似文献
20.
Cselenyák A Benko Z Szepes M Kiss L Lacza Z 《Journal of visualized experiments : JoVE》2011,(57):e3575
Stem cell transplantation protocols are finding their way into clinical practice. Getting better results, making the protocols more robust, and finding new sources for implantable cells are the focus of recent research. Investigating the effectiveness of cell therapies is not an easy task and new tools are needed to investigate the mechanisms involved in the treatment process. We designed an experimental protocol of ischemia/reperfusion in order to allow the observation of cellular connections and even subcellular mechanisms during ischemia/reperfusion injury and after stem cell transplantation and to evaluate the efficacy of cell therapy. H9c2 cardiomyoblast cells were placed onto cell culture plates. Ischemia was simulated with 150 minutes in a glucose free medium with oxygen level below 0.5%. Then, normal media and oxygen levels were reintroduced to simulate reperfusion. After oxygen glucose deprivation, the damaged cells were treated with transplantation of labeled human bone marrow derived mesenchymal stem cells by adding them to the culture. Mesenchymal stem cells are preferred in clinical trials because they are easily accessible with minimal invasive surgery, easily expandable and autologous. After 24 hours of co-cultivation, cells were stained with calcein and ethidium-homodimer to differentiate between live and dead cells. This setup allowed us to investigate the intercellular connections using confocal fluorescent microscopy and to quantify the survival rate of postischemic cells by flow cytometry. Confocal microscopy showed the interactions of the two cell populations such as cell fusion and formation of intercellular nanotubes. Flow cytometry analysis revealed 3 clusters of damaged cells which can be plotted on a graph and analyzed statistically. These populations can be investigated separately and conclusions can be drawn on these data on the effectiveness of the simulated therapeutical approach. 相似文献