Microarray Profiling and Co-Expression Analysis of Long Noncoding RNA and Apoptosis-Associated mRNA in Mouse Intestinal Mucosa Following IntestinalI schemia/Reperfusion Injury / 中山大学学报(医学科学版)

@#【Objective】To investigate the lncRNA expression profile and potential roles in mouse intestinal mucosa after I/R treatment and explore the co-expression relationship between dysregulated lncRNA and apoptotic mRNA at the early stage of reperfusion. 【Methods】 The expression profiles of lncRNA were obtained using microarray and some lncRNA were further validated by quantitative real- time polymerase chain reaction （qRT- PCR）. Gene ontology（GO）analyses were performed to determine closely related biological functions，especially apoptosis-related functions. Finally， the known apoptosis- related mRNA with obviously changes were selected to construct the co-expression network of the dysregulated lncRNA and their correlated apoptotic mRNA， and were analyzed by CNC analysis to calculate the significant correlation of IncRNA-mRNA pairs.【Results】Compared with sham operation group，the expression profile of lncRNA in intestinal epithelium of mice after intestinal I/R was significantly changed ，including 1 503 up- regulated lncRNAs and 2 099 down- regulated lncRNA （Fold change≥2，P<0.05）. At the same time，1 528 mRNA were up- regulated in I/R group，while 1 630 mRNA were down- regulated（fold change≥2.0，P<0.05）. GO enrichment analysis showed that the main functions involved in regulation were lipid metabolism，redox reaction，stress reaction，apoptosis process，programmed cell death，cell cycle，inflammatory response，endothelial cell differentiation and proliferation， tissue remodeling，MAPK，Wnt，vascular endothelial growth factor signaling pathway and so on. Apoptosis-related subitems were enriched in the up- and down-regulated annotations of GO molecular function in different forms ，which were in the forefront. There was a significant co-expression relationship between apoptosis- related mRNA and dysregulated lncRNA. 【Conclusion】 In this study，we established and preliminarily validated the expression profiles of the differentially expressed lncRNA at the early stage of reperfusion in mouse intestinal ischemia injury. Bioinformatics analysis showed that the important biological function of dysregulated lncRNA was the regulation of apoptosis-related processes，and a large number of those lncRNA were indeed highly coexpressed with apoptotic genes ，which would provide a basis and direction for future research.

Proteomics study of intestinal mucosa after ischemic preconditioning against intestinal ischemic reperfusion injury in rats / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To identify associated proteins involved in the molecular response of ischemic preconditioning (IPC) against intestinal ischemia/reperfusion (II/R) in the intestinal mucosa of rats.</p><p><b>METHODS</b>Sixteen SD rats were randomly divided into II/R and IPC groups. II/R injury in rats was produced by clamping superior mesenteric artery for 60 min followed by 60 min reperfusion. IPC was elicited by 20 min ischemia and 5 min reperfusion before index ischemia. The intestinal mucosa was scratched immediately after 60 min of reperfusion and total proteins were separated by immobilized pH gradient (IPG) based on two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed using Image Master 2D Elite 5.0 image analysis software and identified by MALDI-TOF-MS. The biological information of these proteins was searched in the database of these peptide mass finger-printing (PMF). Western blotting and RT-PCR were used to validate the differentially expressed proteins.</p><p><b>RESULTS</b>Image analysis revealed that averages of 1404+/-20 and 1338+/-20 were detected in II/R and IPC groups. A total of 10 spots yielded good spectra, and 8 spots matched with known proteins after database searching. These proteins were mainly involved in anti-oxidation, inhibiting apoptosis and energy metabolism. Western blot confirmed up-regulation of aldehyde dehydrogenase and RT-PCR confirmed up-regulation of aldose reductase in IPC group.</p><p><b>CONCLUSION</b>The clues provided by comparative proteome strategy will shed light on molecular mechanisms of IPC against II/R injury.</p>

Study on sini decoction in treatment of intestinal ischemia-reperfusion injury in rats: mechanism relating to oxygen radical and bcl-2 protein / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of Sini decoction (SND) on intestinal mucosa in rats with intestinal ischemia reperfusion (I/R) and the mechanism relating to oxygen radical and Bcl-2 protein expression.</p><p><b>METHOD</b>Thirty-two SD rats of both sexes were randomly divided into 4 equal groups: (1) control group in which sham operation was performed; (2) model group in which intestinal I/R was produced by clamping super mesenteric artery(SMA) for 1 hour and declamping SMA for 3 hours; (3) SND low dose group in which SND(3 g x kg(-1) x d(-1)) was given via stomach tube for 3 days before operation; (4) SND high dose group in which SND(6 g x kg(-1) x d(1)) was given via stomach tube, for 3 days before operation. A strip of small intestine was taken from distal end of ileum for light microscopic examination, Chiu's score and the detection of intestinal water content (IWC). Apoptosis of intestinal mucosa cell was examined by TUNEL method. Superoxide dismutase (SOD) activity, malondisldehyde (MDA) concentration of intestinal mucosa were detected. The protein express of Bcl-2 of intestinal mucosa was analyzed by immunochemistory.</p><p><b>RESULT</b>Intestinal /R resulted in histopathological changes in intestinal mucosa, increased Chiu's scores, apoptosis index, IWC and MDA content, and reduced SOD activity and the protein expression of Bcl-2 significantly (P < 0.01). The pretreatment of SND could attenuate the above changes significantly (P < 0.01), but there was no significant difference for the above variables between SND low dose group and SND high dose group (P > 0.05). Apoptosis index was significantly negatively correlative to SOD activity in model group and two SND groups. There were significantly negative correlation between apoptosis index and protein expression of Bcl-2 in model group and SND low dose group.</p><p><b>CONCLUSION</b>SND can attenuate intestinal mucosa injury following intestinal U/R, which is related to reducing intestinal mucosa cell apoptosis by removing oxygen free radical and upregulating the protein expression of Bcl-2.</p>

Role of NF-kappaB in the induction of delayed preconditioning in rat heart by pretreatment with Sini decoction / 中国中药杂志

<p><b>OBJECTIVE</b>The present study was designed to investigate whether Sini decoction (SND) containing several chinese medicinal herbs, could induce delayed preconditioning-like effect in rat heart and the possible mechanism.</p><p><b>METHOD</b>In anesthetized open-chest Sprague Dawley rats, left anterior descending coronary artery (LAD) was occluded for 1 h and reperfused vided into four groups: Sham ( n = 8) group in which the surgical procedure was identical to other groups,but the LAD ligature was not ligated; I/R group (n = 8) in which the LAD of rat hearts was subjected to the index occlusion; IPC/SWOP group (n = 8) in which the LAD was occluded for three cycles of 5-minute episodes of preconditioning ischemia followed by 5-minute episodes of reperfusion 24 hours prior to the index occlusion; SND group (n = 8) in which rats were pretreated with Sini decoction (5 g x kg(-1) d(-1)) for three days, the last treatment was pretreated 24 h before the index occlusion. Myocardial infarct size, the activity of superoxide dismutase( SOD) and the content of malondialdehyde (MDA) in myocardial tissue were measured. The mRNA expression of MnSOD and Cu-ZnSOD were detected by RT-PCR. The expression of P65 subunit of NF-kappaB was determined by immunohistochemistry assay.</p><p><b>RESULT</b>As compared with I/R group, myocardial infarct size and the amount of MDA in myocardial tissue were significantly decreased, the activity of SOD and the mRNA expression of MnSOD were increased, the translocation of NF-kappaB was detected and furthermore the expression of NF-kappaB was significantly elevated in SND group as well as in IPC/SWOP group. The mRNA expression of Cu-ZnSOD was lowered in I/R, SND and IPC/SWOP groups, and there was no significant difference among these groups.</p><p><b>CONCLUSION</b>The results suggest that Sini decoction can induce delayed preconditioning-like effect in rat heart and the activation of NF-kappaB may play an important role in it.</p>

Effects of Astragalus membranaceus injection on nitric oxide and endothelin concentration of intestinal mucosa after hemorrhage shock-reperfusion in rats / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effects of Stragalus membranaceus injection on nitric oxide and endothelin levels of intestinal mucosa in reperfusion injury after hemorrhage shock.</p><p><b>METHOD</b>32 SD rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with Astragalus membranaceus 10 g x kg(-1)); high dosage group (treated with Astragalus membranaceus 20 g x kg(-1)). Models of hemorrhagic shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology was observed, and the concentration of lactic acid (LD), nitric oxide (NO), endothelin (ET) of intestinal mucosa were detected.</p><p><b>RESULT</b>The intestinal pathology showed that intestinal mucosa epithelial cells damage in model group was severe, in low dosage group was medium, in high dosage group was slight, and no obvious damage was found in normal group. The concentration of LD and NO of small intestine mucous membrane in model group and low dosage group were significantly higher than those in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). The concentration of ET of small intestine mucous membrane in model group was the highest of the four groups (P < 0.05). The concentration of ET in low dosage group was significantly higher than that in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05).</p><p><b>CONCLUSION</b>Stragalus membranaceus injection can reduce small intestine mucous damage by protecting endothelium function in injury after hemorrhage shock-reperfusion.</p>