Neuroinflammation is a major pathophysiological factor that results in the development of brain injury after cerebral ischemia/reperfusion.Downregulation of microRNA(miR)-455-5p after ischemic stroke has been consider...Neuroinflammation is a major pathophysiological factor that results in the development of brain injury after cerebral ischemia/reperfusion.Downregulation of microRNA(miR)-455-5p after ischemic stroke has been considered a potential biomarker and therapeutic target for neuronal injury after ischemia.However,the role of miR-455-5p in the post-ischemia/reperfusion inflammatory response and the underlying mechanism have not been evaluated.In this study,mouse models of cerebral ischemia/reperfusion injury were established by transient occlusion of the middle cerebral artery for 1 hour followed by reperfusion.Agomir-455-5p,antagomir-455-5p,and their negative controls were injected intracerebroventricularly 2 hours before or 0 and 1 hour after middle cerebral artery occlusion(MCAO).The results showed that cerebral ischemia/reperfusion decreased miR-455-5p expression in the brain tissue and the peripheral blood.Agomir-455-5p pretreatment increased miR-455-5p expression in the brain tissue,reduced the cerebral infarct volume,and improved neurological function.Furthermore,primary cultured microglia were exposed to oxygen-glucose deprivation for 3 hours followed by 21 hours of reoxygenation to mimic cerebral ischemia/reperfusion.miR-455-5p reduced C-C chemokine receptor type 5 mRNA and protein levels,inhibited microglia activation,and reduced the production of the inflammatory factors tumor necrosis factor-αand interleukin-1β.These results suggest that miR-455-5p is a potential biomarker and therapeutic target for the treatment of cerebral ischemia/reperfusion injury and that it alleviates cerebral ischemia/reperfusion injury by inhibiting C-C chemokine receptor type 5 expression and reducing the neuroinflammatory response.展开更多
Certain microRNAs(miRNAs)can function as neuroprotective factors after reperfusion/ischemia brain injury.miRNA-142-3p can participate in the occurrence and development of tumors and myocardial ischemic injury by negat...Certain microRNAs(miRNAs)can function as neuroprotective factors after reperfusion/ischemia brain injury.miRNA-142-3p can participate in the occurrence and development of tumors and myocardial ischemic injury by negatively regulating the activity of Rac1,but it remains unclear whether miRNA-142-3p also participates in cerebral ischemia/reperfusion injury.In this study,a model of oxygen-glucose deprivation/re-oxygenation in primary cortical neurons was established and the neurons were transfected with miR-142-3p agomirs or miR-142-3p antagomirs.miR-142-3p expression was down-regulated in neurons when exposed to oxygen-glucose deprivation/re-oxygenation.Over-expression of miR-142-3p using its agomir remarkably promoted cell death and apoptosis induced by oxygen-glucose deprivation/re-oxygenation and improved mitochondrial biogenesis and function,including the expression of peroxisome proliferator-activated receptor-γcoactivator-1α,mitochondrial transcription factor A,and nuclear respiratory factor 1.However,the opposite effects were produced if miR-142-3p was inhibited.Luciferase reporter assays verified that Rac Family Small GTPase 1(Rac1)was a target gene of miR-142-3p.Over-expressed miR-142-3p inhibited NOX2 activity and expression of Rac1 and Rac1-GTPase(its activated form).miR-142-3p antagomirs had opposite effects after oxygen-glucose deprivation/re-oxygenation.Our results indicate that miR-142-3p down-regulates the expression and activation of Rac1,regulates mitochondrial biogenesis and function,and inhibits oxygen-glucose deprivation damage,thus exerting a neuroprotective effect.The experiments were approved by the Committee of Experimental Animal Use and Care of Central South University,China(approval No.201703346)on March 7,2017.展开更多
Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute...Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury （RIRI）. Methods： Fifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/（kg.d）], SHT low-dose group [1.5 g/（kg,d）] and SHT high-dose group [3.0 g/（kg.d）], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope. Results： Compared with the sham surgery group, the serum creatinine （SCr） and blood urea nitrogen （BUN） levels were significantly increased, Na＋-K＋-ATPase protein level was decreased, and kidney injury molecule-1 （KIM-1） protein level was increased in the model group after the surgery （P〈0.01 or P〈0.05）. Compared with the model group, the SCr, BUN, pathological scores, Na＋-K＋-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery （P〈0.05 or P〈0.01）. And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group （P〈0.05）. The Iocalizations of Na＋-K＋-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group. Conclusions： The SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.展开更多
Background： Inflammation is supposed to play a key role in the pathophysiological processes of intestinal ischemia-reperlhsion injury （IIRI）, and Candida albicans in human gut commonly elevates inflammatory cytokin...Background： Inflammation is supposed to play a key role in the pathophysiological processes of intestinal ischemia-reperlhsion injury （IIRI）, and Candida albicans in human gut commonly elevates inflammatory cytokines in intestinal mucosa. This study aimed to explore the effect of C. albicans on IIRI. Methods： Fifty female Wistar rats were divided into five groups according to the status of C. a/bicans infection and IIRI operation： group blank and sham; group blank and IIRI; group cetbperazone plus IIR1; group C. albicans plus cetbperazone and IIRI （CCI）; and group C. albicans plus cefbperazone and sham. The levels of inflammatory factors tumor necrosis factor （TNF）-a, interleukin （IL）-6, IL- 1 β, and diamine oxidase （DAO） measured by enzyme-linked immunosorbent assay were used to evaluate the inflammation reactivity as well as the integrity of small intestine. Histological scores were used to assess the mucosal damage, and the C. albicans blood translocation was detected to judge the permeability of intestinal mucosal barrier. Results： The levels of inflammatory factors TNF-a, IL-6, and IL-1β in serum and intestine were higher in rats undergone both C. albicans infection and IIRI operation compared with rats in other groups. The levels of DAO （serum： 44.13 ± 4.30 pg/ml, intestine： 346.21 ± 37.03 pg/g） and Chiu scores （3.41 ± 1.09） which reflected intestinal mucosal disruption were highest in group CCI after the operation. The number ofC. albicans translocated into blood was most in group CCI （[33.80 ± 6.60] x 10-2 colony forming unit （CFU）/ml）. Conclusion： Intestinal C. albicans infection worsened the llRl-induced disruption of intestinal mucosal barrier and facilitated the subsequent C. alhicans translocation and dissemination.展开更多
Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI....Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI.However,the mechanisms underlying the regulatory effect of autophagy in AKI among patients undergoing cardiac surgeries are poorly understood.In this study,we aimed to test the hypothesis that exosomal microRNAs(miRNAs)regulate autophagy in tubular epithelial cells after AKI.Methods:Plasma exosomal RNA was extracted from young and elderly AKI patients undergoing cardiac surgery,and the miRNAs expression during the perioperative period were analyzed using next-generation sequencing.The screened miRNAs and their target genes were subjected to gene oncology function and Kyoto Encyclopedia of Genes and Genome enrichment analyses.Renal tubular epithelial cell line(HK-2 cells)was cultured and hypoxia/reoxygenation(H/R)model was established,which is an in vitro renal ischemia/reperfusion(I/R)model.We used Western blot analysis,cell viability assay,transfection,luciferase assay to investigate the mechanisms underlying the observed increases in the levels of renal I/R injury-mediated exosomal miRNAs and their roles in regulating HK-2 cells autophagy.Results:miR-590-3p was highly enriched in the plasma exosomes of young AKI patients after cardiac surgery.Increased levels of miR-590-3p led to the increases in the expression of autophagy marker proteins,including Beclin-1 and microtubule associated protein 1 light chain 3 beta(LC3II),and prolonged the autophagic response in HK-2 cells after H/R treatment.These effects were achieved mainly via increases in the exosomal miR-590-3p levels,and the tumor necrosis factor receptor-associated factor 6 protein was shown to play a key role in I/R injury-mediated autophagy induction.Conclusion:Exosomes released from HK-2 cells after renal I/R injury regulate autophagy by transferring miR-590-3p in a paracrine manner,which suggests that increasing the miR-590-3p levels in HK-2 cell-derived exosomes may increase autophagy and protect against kidney injury after renal I/R injury.展开更多
Retinal ischemia-reperfusion injury(RIRI)is a common cause of visual impairment and blindness.At the cellular level,ischemic and reperfuion retinal injury consists of a self-reinforcing destructive cascade involving o...Retinal ischemia-reperfusion injury(RIRI)is a common cause of visual impairment and blindness.At the cellular level,ischemic and reperfuion retinal injury consists of a self-reinforcing destructive cascade involving oxidative stress initiated by energy failure,inflammatory reaction,calcium influx,increased glutamatergic stimulation and neuronal depolarisation and apoptosis.A number of animal models and analytical techniques have been used to study the retinal ischemia-reperfusion injury,we now understand much better than ever before in the mechanism of RIRI,and an increase in the therapeutic strategies has been developed experimentally to attenuate the detrimental effects of retinal ischemia-reperfusion injury.Thus far,however,success in the laboratory has not been translated to the clinic.Given the increasing understanding of the events involved in ischemia-reperfusion neuronal injury,it is hoped that clinically effective treatments for retinal ischemia-reperfusion injury will soon be available.展开更多
基金supported by the National Natural Science Foundation of China,Nos.82071283(to QH)and 81671130(to QH)Medical Engineering Cross Research Foundation of Shanghai Jiao Tong University of China,No.YG2017MS83(to QH)from Shanghai Municipal Science and Technology Commission Medical Guidance Science and Technology Support Project of China,No.19411968400(to QYM).
文摘Neuroinflammation is a major pathophysiological factor that results in the development of brain injury after cerebral ischemia/reperfusion.Downregulation of microRNA(miR)-455-5p after ischemic stroke has been considered a potential biomarker and therapeutic target for neuronal injury after ischemia.However,the role of miR-455-5p in the post-ischemia/reperfusion inflammatory response and the underlying mechanism have not been evaluated.In this study,mouse models of cerebral ischemia/reperfusion injury were established by transient occlusion of the middle cerebral artery for 1 hour followed by reperfusion.Agomir-455-5p,antagomir-455-5p,and their negative controls were injected intracerebroventricularly 2 hours before or 0 and 1 hour after middle cerebral artery occlusion(MCAO).The results showed that cerebral ischemia/reperfusion decreased miR-455-5p expression in the brain tissue and the peripheral blood.Agomir-455-5p pretreatment increased miR-455-5p expression in the brain tissue,reduced the cerebral infarct volume,and improved neurological function.Furthermore,primary cultured microglia were exposed to oxygen-glucose deprivation for 3 hours followed by 21 hours of reoxygenation to mimic cerebral ischemia/reperfusion.miR-455-5p reduced C-C chemokine receptor type 5 mRNA and protein levels,inhibited microglia activation,and reduced the production of the inflammatory factors tumor necrosis factor-αand interleukin-1β.These results suggest that miR-455-5p is a potential biomarker and therapeutic target for the treatment of cerebral ischemia/reperfusion injury and that it alleviates cerebral ischemia/reperfusion injury by inhibiting C-C chemokine receptor type 5 expression and reducing the neuroinflammatory response.
基金supported by the National Natural Science Foundation of China,No.81771422(to ZY)
文摘Certain microRNAs(miRNAs)can function as neuroprotective factors after reperfusion/ischemia brain injury.miRNA-142-3p can participate in the occurrence and development of tumors and myocardial ischemic injury by negatively regulating the activity of Rac1,but it remains unclear whether miRNA-142-3p also participates in cerebral ischemia/reperfusion injury.In this study,a model of oxygen-glucose deprivation/re-oxygenation in primary cortical neurons was established and the neurons were transfected with miR-142-3p agomirs or miR-142-3p antagomirs.miR-142-3p expression was down-regulated in neurons when exposed to oxygen-glucose deprivation/re-oxygenation.Over-expression of miR-142-3p using its agomir remarkably promoted cell death and apoptosis induced by oxygen-glucose deprivation/re-oxygenation and improved mitochondrial biogenesis and function,including the expression of peroxisome proliferator-activated receptor-γcoactivator-1α,mitochondrial transcription factor A,and nuclear respiratory factor 1.However,the opposite effects were produced if miR-142-3p was inhibited.Luciferase reporter assays verified that Rac Family Small GTPase 1(Rac1)was a target gene of miR-142-3p.Over-expressed miR-142-3p inhibited NOX2 activity and expression of Rac1 and Rac1-GTPase(its activated form).miR-142-3p antagomirs had opposite effects after oxygen-glucose deprivation/re-oxygenation.Our results indicate that miR-142-3p down-regulates the expression and activation of Rac1,regulates mitochondrial biogenesis and function,and inhibits oxygen-glucose deprivation damage,thus exerting a neuroprotective effect.The experiments were approved by the Committee of Experimental Animal Use and Care of Central South University,China(approval No.201703346)on March 7,2017.
基金Supported by National Science and Major Project(No.2010ZX09102-204)National Natural Sciences Foundation of China(No.81072914 and 81273968)the Medicine and Health Foundation of PLA(No.10ZYZ255)
文摘Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury （RIRI）. Methods： Fifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/（kg.d）], SHT low-dose group [1.5 g/（kg,d）] and SHT high-dose group [3.0 g/（kg.d）], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope. Results： Compared with the sham surgery group, the serum creatinine （SCr） and blood urea nitrogen （BUN） levels were significantly increased, Na＋-K＋-ATPase protein level was decreased, and kidney injury molecule-1 （KIM-1） protein level was increased in the model group after the surgery （P〈0.01 or P〈0.05）. Compared with the model group, the SCr, BUN, pathological scores, Na＋-K＋-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery （P〈0.05 or P〈0.01）. And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group （P〈0.05）. The Iocalizations of Na＋-K＋-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group. Conclusions： The SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.
文摘Background： Inflammation is supposed to play a key role in the pathophysiological processes of intestinal ischemia-reperlhsion injury （IIRI）, and Candida albicans in human gut commonly elevates inflammatory cytokines in intestinal mucosa. This study aimed to explore the effect of C. albicans on IIRI. Methods： Fifty female Wistar rats were divided into five groups according to the status of C. a/bicans infection and IIRI operation： group blank and sham; group blank and IIRI; group cetbperazone plus IIR1; group C. albicans plus cetbperazone and IIRI （CCI）; and group C. albicans plus cefbperazone and sham. The levels of inflammatory factors tumor necrosis factor （TNF）-a, interleukin （IL）-6, IL- 1 β, and diamine oxidase （DAO） measured by enzyme-linked immunosorbent assay were used to evaluate the inflammation reactivity as well as the integrity of small intestine. Histological scores were used to assess the mucosal damage, and the C. albicans blood translocation was detected to judge the permeability of intestinal mucosal barrier. Results： The levels of inflammatory factors TNF-a, IL-6, and IL-1β in serum and intestine were higher in rats undergone both C. albicans infection and IIRI operation compared with rats in other groups. The levels of DAO （serum： 44.13 ± 4.30 pg/ml, intestine： 346.21 ± 37.03 pg/g） and Chiu scores （3.41 ± 1.09） which reflected intestinal mucosal disruption were highest in group CCI after the operation. The number ofC. albicans translocated into blood was most in group CCI （[33.80 ± 6.60] x 10-2 colony forming unit （CFU）/ml）. Conclusion： Intestinal C. albicans infection worsened the llRl-induced disruption of intestinal mucosal barrier and facilitated the subsequent C. alhicans translocation and dissemination.
基金National Natural Science Foundation of China(No. 81970344)
文摘Background:Acute kidney injury(AKI)is a common complication in patients,especially elderly patients,who undergo cardiac surgery with cardiopulmonary bypass.Studies have indicated a protective role of autophagy in AKI.However,the mechanisms underlying the regulatory effect of autophagy in AKI among patients undergoing cardiac surgeries are poorly understood.In this study,we aimed to test the hypothesis that exosomal microRNAs(miRNAs)regulate autophagy in tubular epithelial cells after AKI.Methods:Plasma exosomal RNA was extracted from young and elderly AKI patients undergoing cardiac surgery,and the miRNAs expression during the perioperative period were analyzed using next-generation sequencing.The screened miRNAs and their target genes were subjected to gene oncology function and Kyoto Encyclopedia of Genes and Genome enrichment analyses.Renal tubular epithelial cell line(HK-2 cells)was cultured and hypoxia/reoxygenation(H/R)model was established,which is an in vitro renal ischemia/reperfusion(I/R)model.We used Western blot analysis,cell viability assay,transfection,luciferase assay to investigate the mechanisms underlying the observed increases in the levels of renal I/R injury-mediated exosomal miRNAs and their roles in regulating HK-2 cells autophagy.Results:miR-590-3p was highly enriched in the plasma exosomes of young AKI patients after cardiac surgery.Increased levels of miR-590-3p led to the increases in the expression of autophagy marker proteins,including Beclin-1 and microtubule associated protein 1 light chain 3 beta(LC3II),and prolonged the autophagic response in HK-2 cells after H/R treatment.These effects were achieved mainly via increases in the exosomal miR-590-3p levels,and the tumor necrosis factor receptor-associated factor 6 protein was shown to play a key role in I/R injury-mediated autophagy induction.Conclusion:Exosomes released from HK-2 cells after renal I/R injury regulate autophagy by transferring miR-590-3p in a paracrine manner,which suggests that increasing the miR-590-3p levels in HK-2 cell-derived exosomes may increase autophagy and protect against kidney injury after renal I/R injury.
文摘Retinal ischemia-reperfusion injury(RIRI)is a common cause of visual impairment and blindness.At the cellular level,ischemic and reperfuion retinal injury consists of a self-reinforcing destructive cascade involving oxidative stress initiated by energy failure,inflammatory reaction,calcium influx,increased glutamatergic stimulation and neuronal depolarisation and apoptosis.A number of animal models and analytical techniques have been used to study the retinal ischemia-reperfusion injury,we now understand much better than ever before in the mechanism of RIRI,and an increase in the therapeutic strategies has been developed experimentally to attenuate the detrimental effects of retinal ischemia-reperfusion injury.Thus far,however,success in the laboratory has not been translated to the clinic.Given the increasing understanding of the events involved in ischemia-reperfusion neuronal injury,it is hoped that clinically effective treatments for retinal ischemia-reperfusion injury will soon be available.
