The present study aimed to investigate the change of cytochrome c in postconditioning-attenuated ischemia-reperfusion (I/R)-induced mucosal apoptosis in rat intestine compared with ischemic preconditioning (IPC). Using rat model of intestine I/R injury, male Sprague-Dawley rats weighing 220-250 g were divided into 4 groups which were Sham operation group, I/R group, IPC group and ischemic postconditioning (IPOST) group. In these groups, I/R procedure was performed by the occlusion of the superior mesenteric artery (SMA) for 45 min followed by reperfusion for 1 h. In Sham group, there was no intervention. In IPC group, SMA was occluded for 5 min and reperfused for 5 min, for two cycles, before the prolonged occlusion. In IPOST group, three cycles of 30-s reperfusion and 30-s reocclusion were preceded at the start of reperfusion. After the reperfusion, the small intestines were sampled for experimental detection. Intestinal mucosal mitochondrial membrane potential was detected by confocal laser scanning microscopy. Expressions of cytochrome c and caspase-3 proteins were detected using Western-blot method. The apoptosis of intestinal mucosal cells was determined with agarose gel electrophoresis and deoxynucleotidyl transferase mediated dUTP-biotin nick-end labeling (TUNEL) technique. Compared with I/R group, the mitochondrial membrane potentials and the expressions of cytochrome c protein were significantly increased, while the expressions of caspase-3 and the apoptotic rates were decreased in IPOST and IPC groups (P<0.05). There were no significant differences between IPOST and IPC groups (P>0.05). These data provide substantial evidence that IPOST attenuates I/R-induced mucosal apoptosis by reducing the release of cytochrome c from mitochondria in the rat small intestine.
Animals ; Apoptosis ; physiology ; Cytochromes c ; metabolism ; Intestinal Mucosa ; metabolism ; pathology ; Intestines ; blood supply ; Ischemic Postconditioning ; methods ; Male ; Membrane Potential, Mitochondrial ; Mitochondria ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control

Animals ; Colon ; blood supply ; pathology ; Colonic Diseases ; drug therapy ; etiology ; physiopathology ; Hemodynamics ; Hepatic Veins ; pathology ; physiopathology ; Hypertension, Portal ; complications ; physiopathology ; Intestinal Mucosa ; blood supply ; drug effects ; pathology ; Liver Cirrhosis, Experimental ; complications ; Losartan ; administration & dosage ; therapeutic use ; Male ; Microscopy ; Portal Pressure ; drug effects ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo investigate the expression of R-spondin1 (RSpo1) in the intestinal epithelium of mice with intestinal ischemia-reperfusion injury and explore its significance.

METHODSFifty normal male Kunming mice were randomized into sham-operated group (n=10) and intestinal ischemia-reperfusion injury group (n=40), and in the latter group, the mice were subjected to 20-min intestinal mesenteric artery occlusion followed by reperfusion for 6, 12, 24, or 48 h. Enzyme-linked immunosorbent assay (ELISA) and RT-PCR were used to detect intestinal RSpo1 expression of the mice.

RESULTSThe results of RT-PCR and ELISA showed that RSpo1 expression was significantly decreased in mice at 6 h of reperfusion following the intestinal ischemia (P<0.05), and increased gradually with prolonged repersuion time, reaching the peak level at 24 h (P<0.05). The expression underwent rapid decrease afterwards to a significantly lower level than that in the control group at 48 h (P<0.05).

CONCLUSIONIntestinal ischemia-reperfusion injury may inhibit expression of RSpo1 in the early stage, and enhance its expression in the middle stage. RSpo1 can promote proliferation and differentiation of intestinal epithelial stem cells and plays an important role in the repair intestinal mucosal damage.

Animals ; Cell Proliferation ; Intestinal Mucosa ; cytology ; metabolism ; Intestines ; blood supply ; metabolism ; Male ; Mice ; Random Allocation ; Reperfusion Injury ; metabolism ; Stem Cells ; cytology ; Thrombospondins ; genetics ; metabolism

OBJECTIVETo observe the effect of Qihuang Decoction (QHD) on epithelial cell apoptosis of ischemia-reperfusion (I/R) injured intestinal mucosa in rat.

METHODSThe I/R injured intestinal mucosa rat model was established by clamping superior mesenteric artery (SMA) for 45 min and reperfusing for 60 min. The pathomorphological Changes and epithelial cell apoptosis in the injured intestinal mucosa were observed and compared among groups: the sham-operated group (A), the model group (B), the glutamine treated group (C) and the QHD treated group (D).

RESULTSpathomorphological examination showed that in group A, the intestinal villus was intact; in group B, the intestinal subepithelial space were dilated, and showed evident cleavage between the epithelial top and the lamina propria with bare capillaries, bleeding and ulceration; in group C and D, the above-mentioned pathomorphological changes were alleviated to some extents, appeared only in part of the villa, and the alleviation was more significant in group D than in group C. Chiu's scoring showed that the lowest score (zero) presented in group A and the highest presented in group B; scores in group C and D was significantly lower than that in group B (P < 0.05), but showed insignificant difference between the two groups (P > 0.05). Epithelial cell apoptosis detection showed that the least apoptosis rate presented in group A, and the highest in the group B; while in the group C, it lied between group A and B (all P < 0.05), and showed no statistical significance to group D (P > 0.05), though appeared a lowering trend.

CONCLUSIONQHD could reduce the I/R injured intestinal epithelial mucosa, and its protective mechanism may be related to the inhibition on apoptosis of intestinal mucosal epithelial cells.

Animals ; Apoptosis ; drug effects ; Astragalus Plant ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; Intestinal Mucosa ; blood supply ; drug effects ; pathology ; Male ; Rats ; Rats, Wistar ; Reperfusion Injury ; pathology

OBJECTIVETo observe the effects of Pogostemon cablin (Blanco) Benth. (PCB), a Chinese aromatic herbal medicine, on serum levels of nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), and membrane fluidity of intestinal epithelial cells (IMC) in rats undergoing lower limbs ischemic reperfusion (I/R), for exploring its action in protecting intestinal barrier and the possible mechanisms, and to seek a new way, viewing from Chinese medicine, for providing the experimental bases of gastrointestinal protection against trauma or surgical operation.

METHODSEighty adult Wistar rats were induced into lower limb I/R model and randomized equally into the model group, the three PCB water extract groups treated respectively with high- (4 g x kg(-1) x d(-10), middle- (3 g x kg(-1) x d(-1)), and low-dose (2 g x kg(-1) x d(-1)) of PCB water extract, and three PCB volatile oil groups treated respectively with high- (4 g x kg(-1) x d(-1)), middle- (3 g x kg(-1) x d(-1)), low-dose (2 g x kg(-1) d(-1) of PCB volatile oil. Besides, 10 healthy rats was allocated in a normal control group. PCB preparation was given via gastric infusion for 5 successive days just before modeling. The serum levels of NO and TNF-alpha were monitored, and the membranous fluidity of IMC at I/R region was determined by fluorescence polarization technique.

RESULTSCompared with the control group, both serum NO and TNF-alpha levels in model rats were significantly higher (P < 0.01), and the fluorescence polarization value (P) of IMC obviously increased at the same time (P < 0.05). As compared with the model group, the serum level of NO and TNF-alpha significantly reduced in all the PCB treated groups (P < 0.01 or P < 0.05). As for the membrane fluidity, significant difference was shown between the model group with low-dose of PCB water extract and middle-dose of PCB volatile oil (P < 0.05).

CONCLUSIONPCB could effectively protect the intestinal barrier function by way of maintaining the membrane fluidity of IMC through regulating the level of NO and TNF-alpha in serum.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; Extremities ; blood supply ; Intestinal Mucosa ; cytology ; Membrane Fluidity ; drug effects ; Nitric Oxide ; blood ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Tumor Necrosis Factor-alpha ; blood

No abstract availble.
Adult ; Arteries/abnormalities ; Diagnosis, Differential ; Endoscopy, Gastrointestinal ; Gastrointestinal Hemorrhage/*etiology/pathology ; Humans ; Intestinal Mucosa/blood supply/pathology ; Jejunal Diseases/*etiology/pathology ; Jejunum/*blood supply/pathology ; Male

OBJECTIVETo investigate the influence of cervical sympathetic nerve block (SB) on blood flow volume and barrier function of intestinal mucosa after combined radiation and burn injury in rat.

METHODSSD rats were divided into three groups: control (n = 18), combined injury group (n = 100, rats with Co gamma ray body irradiation with a dose of 5 Gy plus 15% TBSA full-thickness burn injury), and combined injury with SB treatment (n = 100, with the same dose of gamma-ray irradiation and burn injury, treated with SB). Twenty rats were sacrificed on 0, 1, 5, 7 days after combined injuries for various observations. SB was conducted with injection of ropivhydrochloride into the neck bilaterally for the SB group, and same amount of normal saline was injected instead in the combined injury group. Blood flow volume, changes in villus height and crypt depth in jejunum, Na(+)-K+ ATPase activity, permeability of small intestine were measured at different time-points.

RESULTSThe blood flow volume in small intestinal mucosal on 1 post-injury days (PID) [(0.29 +/- 0.07) ml x min(-1) x g(-1)] were obviously decreased than that in normal controls [(1.26 +/- 0.23) ml x min(-1) x g(-1), P < 0.01 ], with serious destruction of pit cells, decrease in intestinal mucosal Na(+)-K+ ATPase activity, and increase in intestinal mucosal permeability. Compared with combined injury group, the blood flow volume was [(0.82 +/- 0.11) ml x min(-1) x g(-1) 1 day after combined injury, P < 0.01], and the Na(+)-K+ ATPase activity was obviously increased, and the permeability of small intestine was ameliorated.

CONCLUSIONSB can increase blood flow volume of rat small intestine after combined radiation and burn injury, promote the repair of intestinal epithelium and improve the barrier function of the intestinal wall.

Animals ; Autonomic Nerve Block ; Blood Volume ; physiology ; Burns ; physiopathology ; Intestinal Mucosa ; blood supply ; metabolism ; physiopathology ; Intestine, Small ; Radiation Injuries, Experimental ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Superior Cervical Ganglion

Acute pancreatitis (AP) is a common acute abdomen in clinic with a rapid onset and dangerous pathogenetic condition. AP can cause an injury of intestinal mucosa barrier, leading to translocation of bacteria or endotoxin through multiple routes, bacterial translocation (BT), gut-origin endotoxaemia, and secondary infection of pancreatic tissue, and then cause systemic inflammatory response syndrome (SIRS) or multiple organ dysfunction syndrome (MODS), which are important factors influencing AP's severity and mortality. Meanwhile, the injury of intestinal mucosa barrier plays a key role in AP's process. Therefore, it is clinically important to study the relationship between the injury of intestinal mucosa barrier and AP. In addition, many factors such as microcirculation disturbance, ischemic reperfusion injury, excessive release of inflammatory mediators and apoptosis may also play important roles in the damage of intestinal mucosa barrier. In this review, we summarize studies on mechanisms of AP.
Acute Disease ; Animals ; Humans ; Intestinal Mucosa ; blood supply ; injuries ; metabolism ; Malnutrition ; pathology ; Microcirculation ; metabolism ; Pancreatitis ; complications ; metabolism ; pathology ; physiopathology ; Reperfusion Injury ; pathology

OBJECTIVETo study the effect of berberine on the release of nitric oxide (NO) by rat intestinal mucous microvascular endothelial cells (RIMECs) cultured in vitro for exploring the pharmacological mechanism of berberine in treating intestinal disease.

METHODSCultured RIMECs in vitro were identified adopting immunofluorescent stain with factor VIII-related antigen. NO level in the supernatant of normal cell culture or cell culture treated with different concentrations of berberine was detected with colorimetry at the 3rd, 6th, 9th and 12th h and compared.

RESULTSCompared with that in the normal control group, NO level increased more obviously in the berberine treated groups, and the best effect was shown in the 5 microg/mL berberine treated group.

CONCLUSIONOne of the important pharmacological mechanisms of berberine might be through promoting the endogenous NO release to induce endothelium-dependent dilatation of microvascular in intestinal mucosa, thus to improve the local microcirculation of intestine.

Animals ; Berberine ; pharmacology ; Cells, Cultured ; Colorimetry ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Intestinal Mucosa ; blood supply ; cytology ; Nitric Oxide ; biosynthesis ; Rats ; Time Factors

OBJECTIVETo investigate the influence of enteral administration of carbachol on the intestinal dysfunction of both severely burn patients and rabbits with partial intestinal ischemia/reperfusion (I/R) injury.

METHODSSeventy-five white rabbits were inflicted with I/R injury and randomized into intestinal I/R (I, n=25), carbachol [C, n=25, with 3g/L carbachol (3 mg/kg) injection into duodenum 1 h after SMA occlusion] and sham operation (SO, n=25, with SMA isolation but no occlusion) groups, and 5 other as normal controls. The blood flow of intestinal mucosa was detected before and after SMA occlusion or admission of carbachol. Changes in diamine oxidase (DAO), D-lactate, xylopyranose absorption, blue dextran discharging time were measured at 2, 4, 6, 8, 24, 48, 72 h after SMA occlusion. In addition, eight severe burn patients with TBSA of 84 +/- 12% were enrolled in the study, and carbachol (15 microg/kg) was administered to patients when abdominal distension or bowel sound was lower than 2 times/min, then the number of abdominal distension and bowel sounds per minute were observed.

RESULTSThe blood flow in intestinal mucosa of rabbits without SMA occlusion was (102 +/- 5) PU, reduced to (48 +/- 6) PU after SMA occlusion, and increased to (77 +/- 3) PU after injection of carbachol. The plasma DAO activity and D-lactic acid content in I group began to increase 4 hours after SMA occlusion, and they reached the peak 24 hours after SMA occlusion (4.63 +/- 0.27 U/ml, 7.9 +/- 2.4 mg/L) , after that they decreased gradually, but still higher than the normal value (0.89 +/- 0.14 U/ml, 2.0 +/- 1.1 mg/L, P < 0.05). In carbachol group, data showed the same trends as that in intestine I/R group with lower values, while no obvious changes were in sham operation group (P > 0.05). The content of D-lactic decreased dramatically 2 hours after D-lactic administration in both I and C groups, increased 6 hours after SMA occlusion, then decreased gradually, but it in C group was always higher than normal values, and little fluctuation was in sham operation group. There was no blue dextran discharge 2 hours after SMA occlusion. The discharging distance increased 6 hours later, but it was obviously shorter than the normal value 24 hrs after operation (P < 0.05) , then it returned to normal 48 to 72 hrs after operation. In the C group, blue dextran discharge was found immediately after its injection, with obvious increase in the discharging distance to peak value (43 +/- 6 cm) 6 hours after injury, and returning to normal (28 +/- 3 cm) gradually. In severe burned patients, the bowel sounds was (1.6 +/- 1.1) per minutes before carbachol administration, then increased dramatically to (6.9 +/- 1.7) per minutes 10 mins after administration, reached to a higher level 30 minutes after administration (8.3 +/- 2.4 ) times/min, and it maintained to (6.1 +/- 1.3) times/min 1 hour after administration. Abdominal distension was ameliorated 2 hours after carbachol administration, six patients were able to defecate.

CONCLUSIONEnteral administration of Carbachol can increase the blood flow of intestine mucosa, help to improve the movement, absorption and barrier functions of intestine, and ameliorate intestinal dysfunction in patients with severe burns.

Adolescent ; Adult ; Animals ; Burns ; drug therapy ; physiopathology ; Carbachol ; therapeutic use ; Disease Models, Animal ; Female ; Humans ; Intestinal Mucosa ; blood supply ; metabolism ; Intestines ; physiopathology ; Male ; Rabbits ; Reperfusion Injury ; drug therapy ; physiopathology