BACKGROUND: Irritable bowel syndrome (IBS) is one of the most common functional intestinal diseases, but its pathogenesis is still unknown. The present study aimed to screen the differentially expressed proteins in the mucosa of colon between IBS with diarrhea (IBS-D) patients and the healthy controls. METHODS: Forty-two IBS-D patients meeting the Rome III diagnostic criteria and 40 control subjects from July 2007 to June 2009 in Chinese PLA General Hospital were enrolled in the present study. We examined the protein expression profiles in mucosa of colon corresponding to IBS-D patients (n = 5) and controls (n = 5) using 2-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS). Secondly, Western blot and immunohistochemical analysis were carried out to validate the screened proteins in 27 IBS-D patients and 27 controls. Thirdly, high-performance liquid chromatography (HPLC) was further carried out to determine ATP concentration in the mucosa of colon between 10 IBS-D patients and 8 controls. Comparisons between 2 groups were performed by Student's t-test or Mann-Whitney U-test. RESULTS: Twelve differentially expressed proteins were screened out. The α-enolase (ENOA) in the sigmoid colon (0.917 ± 0.007 vs. 1.310 ± 0.100, t = 2.643, P = 0.017) and caecum (0.765 ± 0.060 vs. 1.212 ± 0.122, t = 2.225, P = 0.023), Isobutyryl-CoA dehydrogenase (ACAD8) in the sigmoid colon (1.127 ± 0.201 vs. 1.497 ± 0.392, t = 7.093, P = 0.008) of the IBS-D group were significantly lower while acetyl-CoA acetyltransferase (CT) in the caecum (2.453 ± 0.422 vs. 0.931 ± 0.652, t = 8.363, P = 0.015) and ATP synthase subunit d (ATP5H) in the sigmoid (0.843 ± 0.042 vs. 0.631 ± 0.042, t = 8.613,P = 0.007) of the IBS-D group was significantly higher, compared with the controls. The ATP concentration in the mucosa of the sigmoid colon in IBS-D group was significantly lower than that of control group (0.470 [0.180, 1.360] vs. 5.350 [2.230, 7.900], U = 55, P < 0.001). CONCLUSIONS Many proteins related to energy metabolism presented differential expression patterns in the mucosa of colon of the IBS-D patients. The abnormalities in energy metabolism may be involved in the pathogenesis of IBS which deserves more studies to elucidate.
Adenosine Triphosphate ; metabolism ; Adult ; Blotting, Western ; Colon ; metabolism ; pathology ; Diarrhea ; enzymology ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Energy Metabolism ; genetics ; physiology ; Female ; Humans ; Immunohistochemistry ; Intestinal Mucosa ; enzymology ; metabolism ; pathology ; Irritable Bowel Syndrome ; enzymology ; metabolism ; pathology ; Male ; Mass Spectrometry ; Middle Aged ; Proteome ; metabolism

BACKGROUND/AIMS: Doublecortin and CaM kinase-like-1 (DCAMKL1) is a marker of stem cells expressed predominantly in the crypt base in the intestine. However, DCAMKL1-positive cells have been shown to be differentiated tuft cells rather than quiescent progenitors. Tuft cells are the only epithelial cells that express cyclooxygenase 2 (COX-2) in the normal intestinal epithelium. We previously generated Cdx2-transgenic mice as model mice for intestinal metaplasia and gastric carcinoma. In the current study, we investigated the association between COX-2 and DCAMKL1 in gastric carcinoma. METHODS: We examined the association between COX-2 and DCAMKL1 expression in gastric carcinomas in clinical samples (early gastric well-differentiated adenocarcinoma) and Cdx2-transgenic mice; and the DCAMKL1-transgenic mouse stomach using immunohistochemistry and quantitative real-time polymerase chain reaction. RESULTS: The COX-2-expressing cells were scattered, not diffusely expressed, in gastric carcinomas from humans and Cdx2-transgenic mice. DCAMKL1-positive cells were also scattered in the gastric carcinomas, indicating that tuft cells could still be present in gastric carcinoma. COX-2 was expressed in DCAMKL1-positive tuft cells in Cdx2- and DCAMKL1-transgenic mouse stomachs, whereas the Sox9 transcription factor was ubiquitously expressed in gastric carcinomas, including COX-2-positive cells. CONCLUSIONS: COX-2 is expressed in DCAMKL1-expressing quiescent tuft cells in gastric carcinoma.
Adenocarcinoma/metabolism ; Animals ; Cyclooxygenase 2/genetics/*metabolism ; Epithelial Cells/metabolism ; Gastric Mucosa/metabolism ; Humans ; Intestinal Mucosa/cytology/*enzymology/metabolism ; Intracellular Signaling Peptides and Proteins/genetics/*metabolism ; Mice ; Mice, Transgenic ; Protein-Serine-Threonine Kinases/genetics/*metabolism ; SOX9 Transcription Factor/genetics/metabolism ; Stomach Neoplasms/*enzymology/genetics

15-Hydroxyprostaglandin dehydrogenase (15-PGDH) is downregulated during the early stages of colorectal carcinogenesis. The aim of the present study was to investigate the potential role of 15-PGDH in normal-appearing colorectal mucosa as a biomarker for predicting colorectal neoplasms. We obtained paired tumor and normal tissues from the surgical specimens of 32 sporadic colorectal cancer patients. mRNA expression of 15-PGDH was measured using a quantitative real-time PCR assay. We evaluated the association between 15-PGDH mRNA expression in normal-appearing mucosa, the presence of synchronous adenoma, and the cumulative incidence of metachronous adenoma. The relative 15-PGDH expression of normal-appearing mucosa in patients with synchronous adenoma was significantly lower than in patients without synchronous adenoma (0.71 vs 1.00, P = 0.044). The patients in the lowest tertile of 15-PGDH expression in normal-appearing mucosa were most likely to have synchronous adenoma (OR: 10.5, P = 0.024). Patients with low 15-PGDH expression in normal-appearing mucosa also demonstrated more advanced stage colorectal cancer (P = 0.045). However, there was no significant difference in the cumulative incidence of metachronous adenoma according to 15-PGDH mRNA expression in normal-appearing mucosa (P = 0.333). Hence, 15-PGDH in normal-appearing colorectal mucosa can be a useful biomarker of field effect for the prediction of sporadic synchronous neoplasms.
Aged ; Colorectal Neoplasms/*diagnosis/enzymology/pathology ; Down-Regulation ; Female ; Humans ; Hydroxyprostaglandin Dehydrogenases/genetics/*metabolism ; Intestinal Mucosa/*enzymology ; Male ; Middle Aged ; Neoplasm Staging ; Neoplasms, Multiple Primary/enzymology/pathology ; Neoplasms, Second Primary/enzymology/pathology ; Odds Ratio ; Predictive Value of Tests ; RNA, Messenger/metabolism ; Real-Time Polymerase Chain Reaction ; Risk Factors ; Tumor Markers, Biological/*metabolism

OBJECTIVETo observe the effects of up- or down-regulation of haemoxygenase 1 (HO-1) gene expression on intestinal mucosa injury induced by intra-abdominal hypertension (IAH).

METHODS(1) Reproduction of rat model of up- or down-regulation of HO-1 gene expression. Twenty-four healthy adult Wistar rats were divided into Co-PP (HO-1 specific revulsive) 2.5 mg, Co-PP 5.0 mg, Sn-PP (HO-1 specific inhibitor) 2.5 mg, and control groups according to the random number table, with six rats in each group. Rats in groups Co-PP 2.5 mg and Sn-PP 2.5 mg were respectively given Co-PP 2.5 mg/kg and Sn-PP 2.5 mg/kg by intraperitoneal injection, once every 12 hours for 3 days. The rats in group Co-PP 5.0 mg were intraperitoneally injected with Co-PP 5.0 mg/kg, once a day for 3 days. The rats in control group were treated with equal volume of normal saline by intraperitoneal injection. All rats were sacrificed on post injection day (PID) 4, and intestinal mucosa tissues were collected for determination of HO-1 mRNA expression. Optimal dose of Co-PP was chosen for the following experiment. (2) The influence of up- or down-regulation of HO-1 gene expression on intestinal mucosa injury under IAH condition. Another 24 healthy adult Wistar rats were divided into control, IAH, Co-PP+IAH, and Sn-PP+IAH groups according to the random number table, with six rats in each group. The rats in groups Co-PP+IAH and Sn-PP+IAH were intraperitoneally injected with 2.5 mg/kg Co-PP and 2.5 mg/kg Sn-PP, once every 12 hours for 3 days. Equal volume of normal saline was intraperitoneally injected into the rats in control group, once every 12 hours for 3 days. Then, nitrogen gas pneumoperitoneum was used to establish the model of IAH in rats of the latter three groups on PID 4, with IAP at 20 mm Hg (1 mm Hg = 0.133 kPa) , and it was maintained for 2 hours. Puncture and intubation were performed in rats of control group without inflating nitrogen gas. Jejunal segment in the length of 10-15 cm was harvested for collecting intestinal mucosa tissues to determine the HO-1 mRNA expression and diamine oxidase (DAO) content. Serum obtained from portal vein blood was collected to determine the D-lactate, TNF-α, and IL-6 contents. Another jejunal segment in the length of 1-2 cm was harvested for histopathological examination. Data were processed with one-way analysis of variance and t test.

RESULTS(1) The HO-1 mRNA expression in group Co-PP 2.5 mg was significantly higher than that in control and Co-PP 5.0 mg groups (with t values respectively 4.756, 3.175, P < 0.05 or P < 0.01). The HO-1 mRNA expression in group Sn-PP 2.5 mg was significantly lower than that in control group (t = 4.880, P < 0.01). The optimal dose of Co-PP for the following experiment was 2.5 mg/kg. (2) HO-1 mRNA expression in group Co-PP+IAH was 60 ± 5, and it was obviously higher than that of group IAH (49 ± 5, t = 3.811, P < 0.01) and control group (39 ± 4, t = 8.034, P < .001) . HO-1 mRNA expression was higher in group IAH than in control group (t = 3.826, P < 0.01). HO-1 mRNA expression in group Sn-PP+IAH was 29 ± 4, which was obviously lower than that of control group (t = 4.330, P < 0.01). The contents of DAO and D-lactate in group Co-PP+IAH were (0.52 ± 0.05) U/mL and (1.9 ± 0.6) mg/L, which were significantly lower than those in group IAH [(0.88 ± 0.06) U/mL and (4.3 ± 0.7) mg/L, with t values respectively 11.291, 6.376, P values all below 0.01], but still higher than those in control group [(0.34 ± 0.04) U/mL, (1.2 ± 0.5) mg/L, with t values respectively 6.886, 2.295, P < 0.05 or P < 0.01]. The contents of TNF-α and IL-6 were much lower in group Co-PP+IAH than in group IAH, but still higher than in control group (with t values from 3.781 to 18.557, P values all below 0.01). The contents of DAO, D-lactate, TNF-α, and IL-6 in group Sn-PP+IAH were all higher than those in the other 3 groups (with t values from 4.181 to 32.938, P values all below 0.01). Structure of epithelial cells from intestinal mucosa was intact and regularly arranged in rats of control group. Intestinal mucosal tissue was edematous, and the top of villi was anabrotic and necrotic in rats of group IAH. Compared with that of group IAH, the degree of intestinal mucosa injury was alleviated in rats of group Co-PP+IAH, while the pathology was aggravated in rats of group Sn-PP+IAH.

CONCLUSIONSUp-regulation of HO-1 gene expression can ameliorate intestinal mucosa injury caused by IAH, thus protecting intestinal mucosa tissues.

Animals ; Disease Models, Animal ; Gene Expression Regulation ; Heme Oxygenase (Decyclizing) ; metabolism ; Intestinal Mucosa ; enzymology ; pathology ; Intra-Abdominal Hypertension ; enzymology ; pathology ; Rats ; Rats, Wistar ; Up-Regulation

OBJECTIVETo investigate the protective effect of baicalin on the intestinal mucosal injury caused by endotoxin-lipopolysaccharide (LPS) and the anti-oxidative injury in colonic and ileal mucosa of rats with septicopyemia.

METHODFifty healthy male BALB/c mice were randomly divided into 5 groups: the normal control group, the model group, and baicalin high-dose, medium-dose and low-dose groups. They were orally administered with double distilled water, 100 mg x kg(-1) of baicalin, 50 mg x kg(-1) of baicalin, and 25 mg x kg(-1) of baicalin respectively for three days, once a day. 1 h after the oral administration on 3 d, they were intraperitoneally injected with normal saline or LPS (17 mg x kg(-1)). At 20 h after the injection of LPS, all of the mice were sacrificed, and their colonic and ileal tissues were collected. The mental status, life state and death rate of mice in each group were observed, and the lengths of colonic were measured. Chiu's scoring method was used to assess the intestinal mucosal injury. Histopathological changes of intestinal tissues were tested by HE staining. The ultraviolet spectrophotometry was used to detect total antioxidant capacity (T-AOC), superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-PX) of intestinal homogenate. The immunohistochemical method was used to analyze the expression of PCNA in intestinal tissues of each group.

RESULTThe death of mice was observed after the intraperitoneal injection of LPS. The death rates of baicalin groups were remarkably lower than the death rate of the model group. The colons in the medium-dose baicalin group were much longer than that in the model group (P < 0.05), with a much lower intestinal mucosa injury degree than the model group. Colonic and ileal injuries in the high-dose baicalin group significantly (P < 0.05). Colonic and ileal injuries in the medium-dose baicalin group and the low-dose baicalin group significantly reduced compare with the model group (P < 0.000 1). The medium-dose baicalin group showed no significant increase in homogenate's T-AOC, T-SOD and GSH-PX compare with the model group (P < 0.05). There was no significant difference between baicalin groups and the model group in PCNA.

CONCLUSIONBaicalin can protect intestinal epithelial cells suffering from injury from oxygen radicals, and relieve the intestinal injury caused by LPS by improving the intestinal mucosa structure and functions.

Animals ; Antioxidants ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Flavonoids ; pharmacology ; Glutathione Peroxidase ; metabolism ; Humans ; Ileum ; drug effects ; enzymology ; injuries ; Intestinal Mucosa ; drug effects ; enzymology ; injuries ; Lipopolysaccharides ; adverse effects ; Male ; Mice ; Mice, Inbred BALB C ; Protective Agents ; pharmacology ; Sepsis ; drug therapy ; prevention & control ; Superoxide Dismutase ; metabolism

AIM: The most important side effect of methotrexate (MTX) is mucositis. The purpose of this study was to evaluate the effect of turmeric extract on intestinal damage and oxidative stress in rats receiving methotrexate. METHODS: Experiments were performed on male Wistar albino rats divided into six groups. First group received normal saline orally, the second group received turmeric extract (100 mg·kg(-1)) orally for 30 days, the third group received turmeric extract (200 mg·kg(-1)) orally for 30 days, the fourth group received a single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30, the fifth group received turmeric extract (100 mg·kg(-1)) orally for 30 days and a single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30, and the sixth group received turmeric extract (200 mg·kg(-1)) orally for 30 days and single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30. Four days after methotrexate injection, animals were anesthetized, blood samples were taken to determine total antioxidant status (TAS) and jejunum samples were taken for glutathione peroxidase (GPx), superoxidase dismutase (SOD), catalase (CAT), aldehyde malondialdehyde (MDA), and histopathological assessment. RESULTS: Microscopic evaluation from intestinal tissues of the MTX treated group, showed severe villus shortening and blunting, inflammatory cell infiltration and hemorrhage in lamina propria, along with epithlial cell necrosis. Levels of SOD, GSH-Px and CAT decreased in the MTX received group, but increased significantly (P < 0.05) in the turmeric + MTX groups. MTX increased lipid peroxidation, however, turmeric decreased peroxidation significantly (P < 0.05). CONCLUSION These results suggest that turmeric extract may protect the small intestine of rats from methotrexate-induced damage. Turmeric effects could result from its antioxidant properties.
Animals ; Catalase ; metabolism ; Curcuma ; chemistry ; Glutathione Peroxidase ; metabolism ; Humans ; Intestinal Diseases ; chemically induced ; drug therapy ; enzymology ; metabolism ; Intestinal Mucosa ; metabolism ; Male ; Malondialdehyde ; metabolism ; Methotrexate ; adverse effects ; Oxidative Stress ; drug effects ; Plant Extracts ; administration & dosage ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

OBJECTIVEIn this study, a growing rat model of zinc deficiency was established to investigate the effect of zinc deficiency on intestinal mucosal morphology and digestive enzyme activity as well as to provide a scientific basis for zinc supplementation therapy in patients with diarrhea.

METHODThree-week-old weaned Sprague-Dawley male rats (n = 30) were randomly divided into 3 groups with 10 in each: rats in the control group (ZA) were fed with a normal diet containing 30 µg/g zinc; rats in the zinc deficient group (ZD) were fed with a zinc-deficient diet containing 0.4 µg/g zinc (refer to AIN-76 formula); and rats in the paired fed group (PF) were fed with a normal diet, but the food intake was limited to intake of rats in ZD group in the previous day. All rats were provided with deionized water for drinking. Their body weight was measured and the food intake during the previous day was recorded early in the morning of the following day. Symptoms of zinc deficiency, such as anorexia, diarrhea, dermatitis, and growth retardation, were observed. Two weeks later, the rats were sacrificed and serum zinc concentration was measured. Jejunal mucosa was taken for biopsy and was stained with hematoxylin and eosin (HE). The height ratio of the jejunal mucosal villi and crypts was measured. In addition, the activity of lactase in the jejunal mucosal brush border, γ-glutamyl peptidase (GGT), and aminopeptidase N (APN) were measured.

RESULTThe average weight of the rats in the ZA, ZD, and PF groups at the beginning of the experiment was (67.4 ± 5.3) g, (64.7 ± 4.8) g, and (66.5 ± 4.1) g, respectively, and the average daily food intake was (11.2 ± 1.0) g, (11.6 ± 1.6) g, and (11.2 ± 1.4) g, respectively. The intergroup differences were not significant. On the 7(th) day of experiment, no significant differences in average food intake were observed between the ZD group and the ZA and PF groups, but the average body weight in the ZD group was significantly lower than that in the ZA and PF groups (P < 0.01). At the end of the experiment (2 weeks), the average weight in the ZD group (112.0 ± 11.5) g was significantly lower than that in the ZA (164.0 ± 15.9) g and PF groups (137.5 ± 16.2) g. The average food intake in the ZD group (13.4 ± 5.1) g was significantly lower than that in the ZA group (18.2 ± 2.4) g (P < 0.01). Serum zinc level in the ZD group (733 ± 231) µg/L was significantly lower than that in the ZA (1553 ± 159) µg/L and PF groups (1457 ± 216) µg/L (P < 0.01). The height ratio of jejunal mucosa villus and crypt in the ZA, ZD, and PF groups was 2.98 ± 0.5, 2.77 ± 0.5, and 2.81 ± 0.7, respectively, and lactase activity was (26.1 ± 15.0) U/mg, (27.4 ± 12.8) U/mg, and (40.8 ± 18.5) U/mg, respectively, without significant intergroup differences. The GGT activity in the jejunal mucosa in the ZD group (12.7 ± 6.5) U/g was significantly lower than that in the ZA (19.1 ± 10.4) U/g and PF groups (18.5 ± 7.7) U/g, but the difference was not significant. The activity of APN in the jejunal mucosa in the ZD group (25.5 ± 7.5) U/g was significantly lower than that in the ZA (48.7 ± 16.8) U/g and PF groups (43.9 ± 14.5) U/g (P < 0.01).

CONCLUSIONZinc deficiency can cause loss of appetite, weight loss, and decreased activity of peptidase in the jejunal mucosal brush border. Zinc deficiency has little effect on the height ratio of the villus and crypt and lactase activity, thereby indicating that zinc deficiency may first affect protein digestion and absorption.

Animals ; Intestinal Mucosa ; enzymology ; metabolism ; pathology ; Jejunum ; metabolism ; pathology ; Lactase ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Zinc ; deficiency

BACKGROUNDMassive blood loss due to trauma is the leading cause of death in trauma patients and military combatants. The fluid category of resuscitation for hypotensive trauma patients is open to debate. This study was conducted to investigate the early effects of hypertonic and isotonic saline solutions on heme oxygenase-1 (HO-1) mRNA expression and apoptosis in the intestinal mucosa of rats with hemorrhagic shock.

METHODSA model of severe hemorrhagic shock was established in 21 Sprague-Dawley rats. The rats were randomly divided into sham, normal saline resuscitation (NS), and hypertonic saline resuscitation (HTS) groups, with 7 in each group. We assessed and compared the HO-1 mRNA expression and apoptosis in the small intestinal mucosa of rats after hemorrhagic shock and resuscitation using the SYBR Green I fluorescence quantitative reverse transcriptase polymerase chain reaction, fluorescein-iso-thiocyanate-annexin V/propidium iodide double staining, and flow cytometry.

RESULTSIn the early stage of hemorrhagic shock and resuscitation, marked apoptosis occurred in the small intestinal mucosa from both the NS and HTS groups. The apoptotic rate in the NS group was higher than that in the HTS group (P < 0.01). Among the three groups, HO-1 mRNA mucosa from the HTS group had the highest level of expression; however, the differences were not significant. There was a significant negative correlation between HO-1 mRNA expression and apoptosis in the small intestinal mucosa from the NS and HTS groups after hemorrhagic shock and resuscitation.

CONCLUSIONSIn this rat model of severe hemorrhagic shock, HTS resuscitation with a small volume is more effective than NS resuscitation in reducing apoptosis of the intestinal mucosa. Further, HO-1 mRNA over-expression in the intestinal mucosa may be one of the molecular mechanisms of HTS in the resuscitation of hemorrhagic shock.

Animals ; Apoptosis ; drug effects ; Flow Cytometry ; Heme Oxygenase-1 ; genetics ; Intestinal Mucosa ; cytology ; drug effects ; Male ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Saline Solution, Hypertonic ; therapeutic use ; Shock, Hemorrhagic ; drug therapy ; enzymology

BACKGROUND/AIMS: Nuclear factor-kappa B p65 (NF-kappa B p65), nuclear factor-kappa B1 p50 (NF-kappa B p50) have been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. Recently, p38 mitogen-activated protein kinase (MAPK)/ NF-kappa B/ cyclin D1 signaling pathway has been shown to play an important part in the pathogenesis of human cancers. This study was designed to investigate the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins in premalignant lesions of colon and colorectal adenocarcinoma. METHODS: Paraffin sections of 20 normal mucosa, 20 low-grade tubular adenoma, 20 high-grade tubular adenoma and 64 adenocarcinoma tissues were analysed immunohistochemically for the expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins. RESULTS: The expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins were significantly higher in adenocarcinoma tissue in comparison with that in normal mucosa, low-grade tubular adenoma, and high-grade tubular adenoma tissues. Expression of NF-kappa B p50 was more frequent in poorly differentiated histologic grade, presence of nodal metastasis, and advanced stage. Expression of p38 MAPK alpha protein was higher in advanced tumor stage, presence of nodal metastasis and advanced stage. Synchronous expression of NF-kappa B p65, NF-kappa B p50, p38 MAPK alpha, and cyclin D1 proteins were significantly higher in adenocarcinoma tissue. CONCULSIONS: With the increased expression of NF-kappa B p65, NF-kappa B p50, and p38 MAPK alpha proteins, p38 MAPK/ NF-kappa B/ cyclin D1 signaling pathway may play a role in the pathogenesis of colorectal carcinoma.
Adenocarcinoma/enzymology/*metabolism/pathology ; Colorectal Neoplasms/enzymology/*metabolism/pathology ; Cyclin D1/immunology/*metabolism ; Data Interpretation, Statistical ; Female ; Humans ; Immunohistochemistry ; Intestinal Mucosa/metabolism ; Male ; Middle Aged ; NF-kappa B/immunology/*metabolism ; NF-kappa B p50 Subunit/immunology/metabolism ; Neoplasm Staging ; Precancerous Conditions/enzymology/*metabolism ; Transcription Factor RelA/immunology/metabolism ; p38 Mitogen-Activated Protein Kinases/*metabolism

OBJECTIVETo investigate the detrimental effects of hemorrhagic shock on the structure and function of mitochondria DNA (mtDNA) encoding cytochrome oxidase genes in intestinal epithelial cells.

METHODSWistar rats were used and divided into two groups: hemorrhagic shock group and control group. Hemorrhagic shock model of rats was utilized in this experiment. The mtDNA was extracted from the intestinal epithelial cells and amplified by polymerase chain reaction (PCR) with different primers of cytochrome oxidase (COX I, COX II and COX III). The products of PCR were directly sequenced.

RESULTSHemorrhagic shock could result in the point mutagenesis in mitochondrial genome encoding cytochrome oxidase (COX I and COX II). There were 4, 4, 22, 16, 35 point mutations in COX I from 5545 to 6838 bp in 5 shocked rats. There were five point mutations in COX II from 7191 to 7542 bp at the site of t7191c, t7212c, a7386g, a7483g, c7542g in 1 shocked rat. There was no mutation found in COX III.

CONCLUSIONSHemorrhagic shock could significantly induce the damage of the gene of cytochrome oxidase encoded by mtDNA.

Animals ; Base Sequence ; DNA, Mitochondrial ; genetics ; Electron Transport Complex IV ; genetics ; Intestinal Mucosa ; enzymology ; Male ; Mutation ; Polymerase Chain Reaction ; Rats ; Rats, Wistar ; Shock, Hemorrhagic ; enzymology ; genetics