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Objective: To detect the levels of angioteinsin II (Ang II) and angioteinsin (1-7) [Ang (1-7)] and the expression levels of angiotensin II type-1 receptor (AT1R) and Mas receptor (MasR) proteins in kidney tissue of the limb ischemia-reperfusion (LIR) mice pre-treated with the angiotensin coverting enzyme 2 (ACE2) activator diminazene (DIZE), and to explore the protective effect of DIZE on the kidney injury of the LIR mice. Methods: Eighteen male ICR mice aged 8 weeks were divided into control group, LIR group and LIR+DIZE group. The mice in model group and LIR+DIZE group were subjected to 2 h of ischemia and 4 h of reperfusion to establish the LIR models. The mice in LIR + DIZE group were pre-treated with 10 mg · kg-1 · d-1 DIZE for 14 d by subcutaneous injection before LIR. The histological technique was used to observe the morphology of kidney tissue of the mice and the pathological injury was evaluated. Chemical colorimetry was performed to determine the levels of serum urea and serum creatinine (Scr) of the mice. Enzyme linked immunosorbent assay (ELISA) was used to determine the Ang II and Ang (1-7) levels in kidney tissue of the mice. Western blotting method was used to measure the expression levels of AT1R and MasR proteins in kidney tissue of the mice. Results: Compared with control group, the pathological changes such as inflammatory cell infiltration and epithelial cell degeneration were found in kidney tissue of the mice in LIR group, and the kindey injury score was obviously increased (P<0. 05); compared with LIR group, the kidney injury performance in the kidey tissue of the mice in LIR + DIZE group was alleviated and the kidney injury score was decreased significantly (P<0. 05). Compared with control group, the levels of serum urea and Scr of the mice in LIR group were significantly increased (P<0. 05); compared with LIR group, the levels of serum urea and Scr of the mice in LIR + DIZE group were significantly decreased (P<0. 05). Compared with control group, the Ang II, Ang (1-7) levels and the ratio of Ang II/Ang (1-7) of the mice in LIR group were significantly increased (P<0. 05); compared with LIR group, the Ang II level of the mice in LIR+DIZE group was markedly decreased (P<0. 05), the Ang (1-7) level was significantly increased (P<0. 05)), and the ratio of Ang II/Ang (1-7) was decreased (P<0.05). Compared with control group, the expression level of AT1R protein in kidney tissue of the mice in LIR group was significantly decreased (P<0.05), the expression of MasR protein was significantly increased (P<0. 05), and the ATlR/MasR ratio was decreased (P<0.05). Compared with LIR group, the AT1R and MasR protein expression levels in kidney tissue of the mice in LIR + DIZE group were significantly increased (P < 0.05), and the ATlR/MasR ratio was also increased (P < 0.05). Conclusion: The imbalance of Ang II/Ang (1-7) and AT1R/Mas expressions in kidney tissue of the mice may be involved in kidney injury after LIR of the mice. ACE2 activitor DIZE may play a protective role in the kidney by improving the imbalance of Ang II/Ang (1-7) and ATlR/MasR expressions.
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Objective To explore the early expression of autophagy associated proteins in lung tissues in acute lung injury in‐duced by remote limb ischemia reperfusion(LIR‐ALI)in Rats .Methods Twelve adult male SD rats weighting 220-250 g were made LIR‐ALI models and divided randomly into two groups(6 in each group):Sham operation group and ischemia‐reperfusion(I/R)group .The rats were anesthetized and the tissues of lung were removed at the end of 4 hours of reperfusion after 3 hours of is‐chemia .The serum lactate dehydrogenase(LDH) were detected with ELISA and the pathological changes of lung tissues were ob‐served by immunofluorescence techniques ;the expression of Beclin1 protein and Atg5 mRNA in the lung tissues were detected by reverse transcription PCR(RT‐PCR) .Microtubules associated protein light chain 3(LC3)in the lung tissues were detected by West‐ern blot test .Results Compared with Sham group ,the level of serum LDH in I/R group were very higher than Sham group(P<0 .01) ,which showed that the rats models of LIR‐ALI were established .The expression of specific antibody ,Beclin1 protein ,Atg5 mRNA and LC3‐Ⅰ ,LC3‐Ⅱ and the ratio of LC3‐Ⅱ /GAPDH in lung tissues were all very higher in I/R group than in Sham group (P<0 .01) .Conclusion ALI can be induced by LIR and the expression of autophagy associated proteins in lung tissues in LIR‐ALI model is very higher in rats .
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<p><b>OBJECTIVE</b>To investigate the effect of H₂S on lower limb ischemia-reperfusion (LIR) induced lung injury and explore the underlying mechanism.</p><p><b>METHODS</b>Wistar rats were randomly divided into control group, IR group, IR+ Sodium Hydrosulphide (NaHS) group and IR+ DL-propargylglycine (PPG) group. IR group as lung injury model induced by LIR were given 4 h reperfusion following 4 h ischemia of bilateral hindlimbs with rubber bands. NaHS (0.78 mg/kg) as exogenous H₂S donor and PPG (60 mg/kg) which can suppress endogenous H₂S production were administrated before LIR, respectively. The lungs were removed for histologic analysis, the determination of wet-to-dry weight ratios and the measurement of mRNA and protein levels of aquaporin-1 (AQP₁), aquaporin-5 (AQP₅) as indexes of water transport abnormality, and mRNA and protein levels of Toll-like receptor 4 (TLR₄), myeloid differentiation primary-response gene 88 (MyD88) and p-NF-κB as indexes of inflammation.</p><p><b>RESULTS</b>LIR induced lung injury was accompanied with upregulation of TLR₄-Myd88-NF-κB pathway and downregulation of AQP1/AQP₅. NaHS pre-treatment reduced lung injury with increasing AQP₁/AQP₅ expression and inhibition of TLR₄-Myd88-NF-κB pathway, but PPG adjusted AQP₁/AQP₅ and TLR4 pathway to the opposite side and exacerbated lung injury.</p><p><b>CONCLUSION</b>Endogenous H₂S, TLR₄-Myd88-NF-κB pathway and AQP₁/AQP₅ were involved in LIR induced lung injury. Increased H₂S would alleviate lung injury and the effect is at least partially depend on the adjustment of TLR₄-Myd88-NF-κB pathway and AQP₁/AQP₅ expression to reduce inflammatory reaction and lessen pulmonary edema.</p>
Subject(s)
Animals , Male , Rats , Acute Lung Injury , Pathology , Aquaporins , Metabolism , Drug Evaluation, Preclinical , Edema , Pathology , Hydrogen Sulfide , Pharmacology , Therapeutic Uses , Inflammation , Lung , Pathology , Myeloid Differentiation Factor 88 , Metabolism , NF-kappa B , Metabolism , Random Allocation , Rats, Wistar , Reperfusion Injury , Pathology , Toll-Like Receptor 4 , Metabolism , Water , MetabolismABSTRACT
The purpose of this study was to investigate the protective effects of ischemic post-conditioning on damage to the barrier function of the small intestine caused by limb ischemia-reperfusion injury. Male Wistar rats were randomly divided into 3 groups (N = 36 each): sham operated (group S), lower limb ischemia-reperfusion (group LIR), and post-conditioning (group PC). Each group was divided into subgroups (N = 6) according to reperfusion time: immediate (0 h; T1), 1 h (T2), 3 h (T3), 6 h (T4), 12 h (T5), and 24 h (T6). In the PC group, 3 cycles of reperfusion followed by ischemia (each lasting 30 s) were applied immediately. At all reperfusion times (T1-T6), diamine oxidase (DAO), superoxide dismutase (SOD), and myeloperoxidase (MPO) activity, malondialdehyde (MDA) intestinal tissue concentrations, plasma endotoxin concentrations, and serum DAO, tumor necrosis factor-α (TNF-α), and interleukin-10 (IL-10) concentrations were measured in sacrificed rats. Chiu’s pathology scores for small intestinal mucosa were determined under a light microscope and showed that damage to the small intestinal mucosa was lower in group PC than in group LIR. In group PC, tissue DAO and SOD concentrations at T2 to T6, and IL-10 concentrations at T2 to T5 were higher than in group LIR (P < 0.05); however, tissue MPO and MDA concentrations, and serum DAO and plasma endotoxin concentrations at T2 to T6, as well as TNF-α at T2 and T4 decreased significantly (P < 0.05). These results show that ischemic post-conditioning attenuated the permeability of the small intestines after limb ischemia-reperfusion injury. The protective mechanism of ischemic post-conditioning may be related to inhibition of oxygen free radicals and inflammatory cytokines that cause organ damage.
Subject(s)
Animals , Male , Rats , Extremities/blood supply , Intestinal Diseases/prevention & control , Intestinal Mucosa/pathology , Intestine, Small/pathology , Ischemic Postconditioning/methods , Reperfusion Injury/prevention & control , Biomarkers/analysis , Intestinal Diseases/pathology , Random Allocation , Rats, Wistar , Reperfusion Injury/pathologyABSTRACT
Objective:To investigate spinal cord neurons fos,heat shock protein 70(HSP70)change and influence of Danshen Injection after the rats’limb ischemia reperfusion.Methods:Through temporarily interdict the rat’s one side iliac artery and femoral artery to establish the model of the limb ischemia reperfusion injury.Using the immunohistochemical method of ABC to observe the expressions about neurons fos and HSP70 positive neurons and the character about their distributions,as well as their change after being intervened by Danshen Injection n.Results:The rats’spinal cord neurons fos and HSP70 expressions increased in four hours later the limb ischemia(P