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Objective To investigate the effect of sevoflurane (Sevo) preconditioning on lung injury induced by ischemia/reperfusion (IR) of hind limbs in rats.Methods Forty-five healthy adult Sprague-Dawley (SD) rats of clean grade were randomly allocated into three groups,15 mice per group.Femoral veins and femoral arteries of rats were isolated but not clipped in Sham operation group (Sham group).Femoral arteries of rats were clipped for ischemia for 3h,followed reperfusion for 3h in ischemia/reperfusion group (IR group).Rats were inhaled with 2.5％ sevoflurane for 30 min,followed establishing the model of IR of hind limbs after 15min in Sevo preconditioning + IR group (SIR group).Rats were euthanized after experimental time out,and left lung tissue was excised.Wet lung weight to dry lung weight (W/D) and total lung water content (TLW) were tested.Pathological changes of lung tissue were observed and index of quantitative evaluation for alveolar damage (IQA) was tested by light microscope,and changes of ultrastructure of lung tissue were observed by transmission electron microscope.The expressions of glucose-regulated protein 78 (GRP78) and CCAAT enhancer binding protein (C/EBP) homologous proteins (CHOP) protein were detected by reverse transcription PCR (RT-PCR) and Western Blot,respectively.Results Compared to Sham group,W/D,TLW and IQA were all notably higher (P ＜ 0.05) in IR group.W/D,TLW and IQA of SIR group were all lower (P ＜ O.05) than those of IR group.The injury of morphological structure and ultrastructure of lung tissue was distinct in IR group but alleviated in SIR group.Compared to Sham group,the expression of GRP78 and CHOP was higher (P ＜ 0.05) in IR group.The expression of GRP78 and CHOP in SIR group was lower (P ＜ 0.05) than that in IR group.Conclusion Sevo preconditioning has protective effect on rats with lung injury induced by IR of hind limbs,which may be related to inhibition of excessive unfolded protein response (UPR).
ABSTRACT
Objective To investigate the effect of sevoflurane (Sevo) preconditioning on lung injury induced by ischemia/reperfusion (IR) of hind limbs in rats.Methods Forty-five healthy adult Sprague-Dawley (SD) rats of clean grade were randomly allocated into three groups,15 mice per group.Femoral veins and femoral arteries of rats were isolated but not clipped in Sham operation group (Sham group).Femoral arteries of rats were clipped for ischemia for 3h,followed reperfusion for 3h in ischemia/reperfusion group (IR group).Rats were inhaled with 2.5％ sevoflurane for 30 min,followed establishing the model of IR of hind limbs after 15min in Sevo preconditioning + IR group (SIR group).Rats were euthanized after experimental time out,and left lung tissue was excised.Wet lung weight to dry lung weight (W/D) and total lung water content (TLW) were tested.Pathological changes of lung tissue were observed and index of quantitative evaluation for alveolar damage (IQA) was tested by light microscope,and changes of ultrastructure of lung tissue were observed by transmission electron microscope.The expressions of glucose-regulated protein 78 (GRP78) and CCAAT enhancer binding protein (C/EBP) homologous proteins (CHOP) protein were detected by reverse transcription PCR (RT-PCR) and Western Blot,respectively.Results Compared to Sham group,W/D,TLW and IQA were all notably higher (P ＜ 0.05) in IR group.W/D,TLW and IQA of SIR group were all lower (P ＜ O.05) than those of IR group.The injury of morphological structure and ultrastructure of lung tissue was distinct in IR group but alleviated in SIR group.Compared to Sham group,the expression of GRP78 and CHOP was higher (P ＜ 0.05) in IR group.The expression of GRP78 and CHOP in SIR group was lower (P ＜ 0.05) than that in IR group.Conclusion Sevo preconditioning has protective effect on rats with lung injury induced by IR of hind limbs,which may be related to inhibition of excessive unfolded protein response (UPR).
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Objective:To investigate mitochondrial-dependent molecular mechanisms of a novel agonistic anti-human death receptor 5 (DR5) monoclonal antibody(mDRA-6) inducing apoptosis in Jurkat cell.Methods:The dose-dependent and time-dependent cell growth suppression of mDRA-6 in Jurkat cells was determined by MTT assay.The measurement of the mitochondrial transmembrane potential(ΔΨm) of Jurkat cells was detected by flow cytometry with JC-1 single staining.Caspase-8,9 as well as Bid,Bax,Bcl-2 and Cyto c of apoptotic Jurkat cells were analyzed by Western blot after mDRA-6 treatment.Results:The mDRA-6 induced cell growth suppression and cytotoxicity in dose-dependent manner and time-dependent manner.After mDRA-6 treatment at 2.0 μg/ml for15 min,30 min,60 min and 120 min,the change in ΔΨm were 20.14 %,19.34 %,21.11% and 30.90% respectively by JC-1 single staining.Western blot revealed that the level of active fragments of Caspase-8,9 and Bid,Bax,Bcl-2 and Cyto c respectively,and the amount of Cyto c was increased in cytosol concomitant with the related attenuation of Cyto c in mitochondria.Conclusion: Apoptotic pathway of Jurkat cells induced by mDRA-6 is initiated upon DR5 ligatian to mDRA-6 and exogenic Caspase-dependent cell apoptotic cascades is activated,and endogenic mitochondrial-dependent cell apoptosis pathway is activated.mDRA-6 may be a useful agent in investigating human leukemia therapy by using TRAIL/DR5.
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Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.
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Objective:To observe anti-DR5 monoclonal antibody on apoptosis and CDC effect in EC109 cells.Methods:The anti-DR5 monoclonal antibody was prepared by hybridoma technique.Tumoricidal effects and complement-dependent cytotoxicity of the McAb to EC109 cells was screened by MTT assay.The apoptosis of EC109 cells was detected by flow cytometry with annexin Ⅴ-FITC/PI staining.Morphological change of EC109 was observed by microphotograph.Results:An anti-DR5 monoclonal antibody was obtained.It induced apoptosis of EC109 dose dependently.The cytotoxic action was notably enhanced by addition of complement.The cells growth inhibition ratios reached 83.04%.The apoptotic body and cathepsis were seen in microphotograph.Conclusion:The anti-DR5 monoclonal antibody could induce EC109 cells apoptosis and cause the complement dependent cytotoxic (CDC) effects powerfully.