AIM: To investigate the role of P-selectin, intercellular adhesion molecule-1 (ICAM-1) and dendritic cells (DCs)in liver/kidney of rats with hepatic/renal ischemiareperfusion injury and the preventive effect of anti-P...AIM: To investigate the role of P-selectin, intercellular adhesion molecule-1 (ICAM-1) and dendritic cells (DCs)in liver/kidney of rats with hepatic/renal ischemiareperfusion injury and the preventive effect of anti-Pselectin lectin-EGF domain monoclonal antibody (anti-PsLEGFmAb) on the injury.METHODS: Rat models of hepatic and renal ischemiareperfusion were established. The rats were then divided into two groups, one group treated with anti-PsL-EGFmAb(n = 20) and control treated with saline (n = 20). Both groups were subdivided into four groups according to reperfusion time (1, 3, 6 and 24 h). The sham-operated group (n = 5) served as a control group. DCs were observed by the microscopic image method, while P-selectin and ICAM-1 were analyzed by immunohistochemistry.RESULTS: P-selectin increased significantly in hepatic sinusoidal endothelial cells and renal tubular epithelial cells 1 h after ischemia-reperfusion, and the expression of ICAM-1 was up-regulated in hepatic sinusoid and renal vessels after 6 h. CD1a+CD80+DCs gradually increased in hepatic sinusoidal endothelium and renal tubules and interstitium 1 h after ischemia-reperfusion, and there was the most number of DCs in 24-h group. The localization of DCs was associated with rat hepatic/renal function.These changes became less significant in rats treated with anti-PsL-EGFmAb.CONCLUSION: DCs play an important role in immune pathogenesis of hepatic/renal ischemia-reperfusion injury.Anti-PsL-EGFmAb may regulate and inhibit local DC immigration and accumulation in liver/kidney.展开更多
Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes ...Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.The gene expression profiles of CD11 b^(+)/Ly6 C^(intermediate)macrophages of the sham surgery mice,and the mice 4 h,24 h and 9 days after renal IRI were downloaded from the Gene Expression Omnibus database.Analysis of m RNA expression profiles was conducted to identify differentially expressed genes(DEGs),biological processes and pathways by the series test of cluster.Protein-protein interaction network was constructed and analysed to discover the key genes.A total of 6738 DEGs were identified and assigned to 20 model profiles.DEGs in profile 13 were one of the predominant expression profiles,which are involved in immune cell chemotaxis and proliferation.Signet analysis showed that Atp5 a1,Atp5 o,Cox4 i,Cdc42,Rac2 and Nhp2 were the key genes involved in oxidation-reduction,apoptosis,migration,M1-M2 differentiation,and proliferation of macrophages.RPS18 may be an appreciate reference gene as it was stable in macrophages.The identified DEGs and their enriched pathways investigate factors that may participate in the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.Moreover,the vital gene Nhp2 may involve the polarization of macrophages,which may be a new target to affect the process of AKI.展开更多
Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis...Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis.However,the regulatory impact of TRPC6 on renal fibrosis remains unclear.Methods In vivo,TRPC6 knockout(TRPC6−/−)mice and age-matched 129 SvEv(WT)mice underwent unilateral renal ischemia-reperfusion(uIR)injury surgery on the left renal pedicle or sham operation.Kidneys and serum were collected on days 7,14,21,and 28 after euthanasia.In vitro,primary tubular epithelial cells(PTECs)were isolated from TRPC6−/−and WT mice,followed by treatment with transforming growth factorβ1(TGFβ1)for 72 h.The anti-fibrotic effect of TRPC6−/−and the underlying mechanisms were assessed through hematoxylin-eosin staining,Masson staining,immunostaining,qRT-PCR,and Western blotting.Results Increased TRPC6 expression was observed in uIR mice and PTECs treated with TGFβ1.TRPC6−/−alleviated renal fibrosis by reducing the expression of fibrotic markers(Col-1,α-SMA,and vimentin),as well as decreasing the apoptosis and inflammation of PTECs during fibrotic progression both in vivo and in vitro.Additionally,we found that the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/glycogen synthase kinase 3 beta(GSK3β)signaling pathway,a pivotal player in renal fibrosis,was down-regulated following TRPC6 deletion.Conclusion These results suggest that the ablation of TRPC6 may mitigate renal fibrosis by inhibiting the apoptosis and inflammation of PTECs through down-regulation of the PI3K/AKT/GSK3βpathway.Targeting TRPC6 could be a novel therapeutic strategy for preventing chronic kidney disease.展开更多
Donation after circulatory-determined death(DCD)is an important part of renal transplantation.Therefore,DCD renal transplantation animal model should be established to study the mechanism of organ injury.Here,we estab...Donation after circulatory-determined death(DCD)is an important part of renal transplantation.Therefore,DCD renal transplantation animal model should be established to study the mechanism of organ injury.Here,we established a stable DCD rat renal transplantation model and investigated the dynamic regulation of graft self-repairing and antioxidant capacities with different non-heart-beating times(NHBTs).Male Sprague-Dawley rats were randomly divided into four groups with the NHBT of the donors from 0 to 15,30,and 45 minutes.Recipients in long NHBT groups had a significantly lower survival rate and poorer graft function than those in short NHBT groups.Grafts from the 15-minute and 30-minute NHBT groups showed light and severe injury respectively at an early stage after transplantation and recovered within 7 days after transplantation,whereas the self-repairing of the grafts in the 45-minute NHBT group was delayed.The expressions of proliferating cell nuclear antigen(PCNA)and von Willebrand factor(vWF)were dependent on NHBT.The expression of antioxidant proteins paralleled graft recovery.In conclusion,the recipients can up-regulate antioxidant capacity to enhance graft self-repairing in DCD renal transplantation.Prolonged NHBT can delay the self-repairing and antioxidation of grafts.展开更多
Objective： To investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet （复方肾华片, SHP） on the expression of Toll-like receptors （TLRs） during renal ischemia-reperfusion ...Objective： To investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet （复方肾华片, SHP） on the expression of Toll-like receptors （TLRs） during renal ischemia-reperfusion injury （IRI）-induced acute kidney injury （AKI） in rats. Methods： A total of 28 Wistar rats were randomly divided into five groups： （1） pseudo-operation control group, （2） ischemia-reperfusion model group, （3） Astragaloside group, （4） high-dose SHP group, and （5） low-dose SHP group. There were four rats in the pseudo-operation group and six rats in each of the other groups. The accepted ischemia-reperfusion model was established after a 7-day gavage intervention, and pathological changes and renal function were observed, using an enzyme-linked immunosorbent assay （ELISA） to detect interleukin 8 （IL-8） and interferon gamma （IFN-r） levels, as well as immunohistochemical staining to detect altered levels of TLR2 and TLR4 expression in renal tissue. Results： After 24 h, renal pathological damage and the expression levels of serum creatinine （Scr）, IL-8, IFN- r, TLR2, and TLR4 were significantly higher in the model group as compared with the pseudo-operation group （P〈0.05）. In addition, at 24 h the above indicators decreased significantly in the Astragaloside group, high- dose SHP group and low-dose SHP group as compared with the ischemia-reperfusion model group （P〈0.05）. TLR2 and TLR4 expression levels were significantly reduced in the SHP treatment and Astragaloside group as compared with the pseudo-operation group （P〈0.05）. Further, the high-dose SHP group showed significantly less renal damage score and decreased levels of TLR expression than those of low-dose SHP group and Astragaloside group （all P〈0.05）. Conclusion： SHP can alleviate the renal structural and functional damage caused by IRI-induced AKI in rats by reducing the damage of renal pathology, which may reduce inflammatory cytokine levels by downregulating the expression of TLRs in renal tissue in a dose-dependent manner.展开更多
肾脏缺血时,坏死和凋亡的肾小管上皮及血管内皮细胞会释放出高迁移率族蛋白(high mobility group box 1,HMGB1)。HMGB1可能是诱导肾脏缺血再灌注损伤(renal ischemia and reperfusion injury,RIRI)早期的内源性危险信号。本研究主要阐...肾脏缺血时,坏死和凋亡的肾小管上皮及血管内皮细胞会释放出高迁移率族蛋白(high mobility group box 1,HMGB1)。HMGB1可能是诱导肾脏缺血再灌注损伤(renal ischemia and reperfusion injury,RIRI)早期的内源性危险信号。本研究主要阐述了在RIRI早期胞外HMGB1的来源以及与RIRI的关系。HMGB1可促使天然调节型T细胞(nTreg)丧失炎症抑制作用甚至可能翻转为促炎性细胞,该分子同时诱导巨噬细胞分泌大量炎性因子,并作用于活化的巨噬细胞使其自分泌HMGB1,二者共同作用造成肾脏缺血再灌注早期炎症损伤。如在损伤早期阻断HMGB1的多重作用,不仅可以消除巨噬细胞的炎症反应,而且可能恢复nTreg的炎症抑制作用从而减轻炎症反应,可能会对肾脏起到更好的保护作用。展开更多
Ischemic postconditioning was defined as rapid intermittent interruptions of blood flow in the early phase of reperfusion,which has been found to be protective against renal ischemia-reperfusion injury(IRI)in animal m...Ischemic postconditioning was defined as rapid intermittent interruptions of blood flow in the early phase of reperfusion,which has been found to be protective against renal ischemia-reperfusion injury(IRI)in animal models but not in clinical trials.We describe a case that the allograft renal vein was twisted because of the surgeon’s mistake,which caused the warm ischemia of allograft after reperfusion.The allograft restored blood flow without second reperfusion and cold preservation after 9 min of warm ischemia.The patient was followed up for 3 months and the allograft worked well without complications.展开更多
基金Supported by Grants from the National Natural Science Foundation of China, No. 39970340the Scientific Fund of the Chinese Ministry of Health, 98-2-283the Natural Science Foundation of Shanghai,No. 02ZB14041 and 034119916
文摘AIM: To investigate the role of P-selectin, intercellular adhesion molecule-1 (ICAM-1) and dendritic cells (DCs)in liver/kidney of rats with hepatic/renal ischemiareperfusion injury and the preventive effect of anti-Pselectin lectin-EGF domain monoclonal antibody (anti-PsLEGFmAb) on the injury.METHODS: Rat models of hepatic and renal ischemiareperfusion were established. The rats were then divided into two groups, one group treated with anti-PsL-EGFmAb(n = 20) and control treated with saline (n = 20). Both groups were subdivided into four groups according to reperfusion time (1, 3, 6 and 24 h). The sham-operated group (n = 5) served as a control group. DCs were observed by the microscopic image method, while P-selectin and ICAM-1 were analyzed by immunohistochemistry.RESULTS: P-selectin increased significantly in hepatic sinusoidal endothelial cells and renal tubular epithelial cells 1 h after ischemia-reperfusion, and the expression of ICAM-1 was up-regulated in hepatic sinusoid and renal vessels after 6 h. CD1a+CD80+DCs gradually increased in hepatic sinusoidal endothelium and renal tubules and interstitium 1 h after ischemia-reperfusion, and there was the most number of DCs in 24-h group. The localization of DCs was associated with rat hepatic/renal function.These changes became less significant in rats treated with anti-PsL-EGFmAb.CONCLUSION: DCs play an important role in immune pathogenesis of hepatic/renal ischemia-reperfusion injury.Anti-PsL-EGFmAb may regulate and inhibit local DC immigration and accumulation in liver/kidney.
基金supported by grants from the National Natural Science Foundation of China(No.81670634)Graduate student scientific research innovation projects in Jiangsu province(No.KYLX15_0981)Nanjing Medical University Science and Technology Development Fund(No.2016NJMU065)
文摘Renal ischemia-reperfusion injury(IRI)is a major cause of acute kidney injury(AKI),which could induce the poor prognosis.The purpose of this study was to characterize the molecular mechanism of the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.The gene expression profiles of CD11 b^(+)/Ly6 C^(intermediate)macrophages of the sham surgery mice,and the mice 4 h,24 h and 9 days after renal IRI were downloaded from the Gene Expression Omnibus database.Analysis of m RNA expression profiles was conducted to identify differentially expressed genes(DEGs),biological processes and pathways by the series test of cluster.Protein-protein interaction network was constructed and analysed to discover the key genes.A total of 6738 DEGs were identified and assigned to 20 model profiles.DEGs in profile 13 were one of the predominant expression profiles,which are involved in immune cell chemotaxis and proliferation.Signet analysis showed that Atp5 a1,Atp5 o,Cox4 i,Cdc42,Rac2 and Nhp2 were the key genes involved in oxidation-reduction,apoptosis,migration,M1-M2 differentiation,and proliferation of macrophages.RPS18 may be an appreciate reference gene as it was stable in macrophages.The identified DEGs and their enriched pathways investigate factors that may participate in the functional changes of CD11 b^(+)/Ly6 C^(intermediate)macrophages after renal IRI.Moreover,the vital gene Nhp2 may involve the polarization of macrophages,which may be a new target to affect the process of AKI.
基金supported by grants from the National Natural Science Foundation of China(Nos.31171087,30970662,and 82100715)the Henan Medical Science and Technology Research Program(Joint Construction,No.LHGJ20190246).
文摘Objective Renal fibrosis is the ultimate pathway of various forms of acute and chronic kidney damage.Notably,the knockout of transient receptor potential channel 6(TRPC6)has shown promise in alleviating renal fibrosis.However,the regulatory impact of TRPC6 on renal fibrosis remains unclear.Methods In vivo,TRPC6 knockout(TRPC6−/−)mice and age-matched 129 SvEv(WT)mice underwent unilateral renal ischemia-reperfusion(uIR)injury surgery on the left renal pedicle or sham operation.Kidneys and serum were collected on days 7,14,21,and 28 after euthanasia.In vitro,primary tubular epithelial cells(PTECs)were isolated from TRPC6−/−and WT mice,followed by treatment with transforming growth factorβ1(TGFβ1)for 72 h.The anti-fibrotic effect of TRPC6−/−and the underlying mechanisms were assessed through hematoxylin-eosin staining,Masson staining,immunostaining,qRT-PCR,and Western blotting.Results Increased TRPC6 expression was observed in uIR mice and PTECs treated with TGFβ1.TRPC6−/−alleviated renal fibrosis by reducing the expression of fibrotic markers(Col-1,α-SMA,and vimentin),as well as decreasing the apoptosis and inflammation of PTECs during fibrotic progression both in vivo and in vitro.Additionally,we found that the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/glycogen synthase kinase 3 beta(GSK3β)signaling pathway,a pivotal player in renal fibrosis,was down-regulated following TRPC6 deletion.Conclusion These results suggest that the ablation of TRPC6 may mitigate renal fibrosis by inhibiting the apoptosis and inflammation of PTECs through down-regulation of the PI3K/AKT/GSK3βpathway.Targeting TRPC6 could be a novel therapeutic strategy for preventing chronic kidney disease.
基金supported by funds from the National Natural Science Foundation of China(Grant No.81570613 and No.81370853)Jiangsu Provincial Social Development Project(Grant No.BE2017615)2016 Jiangsu Provincial Medical Innovation Team(Grant No.0536).
文摘Donation after circulatory-determined death(DCD)is an important part of renal transplantation.Therefore,DCD renal transplantation animal model should be established to study the mechanism of organ injury.Here,we established a stable DCD rat renal transplantation model and investigated the dynamic regulation of graft self-repairing and antioxidant capacities with different non-heart-beating times(NHBTs).Male Sprague-Dawley rats were randomly divided into four groups with the NHBT of the donors from 0 to 15,30,and 45 minutes.Recipients in long NHBT groups had a significantly lower survival rate and poorer graft function than those in short NHBT groups.Grafts from the 15-minute and 30-minute NHBT groups showed light and severe injury respectively at an early stage after transplantation and recovered within 7 days after transplantation,whereas the self-repairing of the grafts in the 45-minute NHBT group was delayed.The expressions of proliferating cell nuclear antigen(PCNA)and von Willebrand factor(vWF)were dependent on NHBT.The expression of antioxidant proteins paralleled graft recovery.In conclusion,the recipients can up-regulate antioxidant capacity to enhance graft self-repairing in DCD renal transplantation.Prolonged NHBT can delay the self-repairing and antioxidation of grafts.
基金Supported by the National Natural Science Foundation of China(No.81072914 and 81273968)Major Project Foundation of National Science and Technology(No.2010ZX9102-204)Traditional Chinese Medicine Research Grant for Military Organization(No.10ZYZ255)
文摘Objective： To investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet （复方肾华片, SHP） on the expression of Toll-like receptors （TLRs） during renal ischemia-reperfusion injury （IRI）-induced acute kidney injury （AKI） in rats. Methods： A total of 28 Wistar rats were randomly divided into five groups： （1） pseudo-operation control group, （2） ischemia-reperfusion model group, （3） Astragaloside group, （4） high-dose SHP group, and （5） low-dose SHP group. There were four rats in the pseudo-operation group and six rats in each of the other groups. The accepted ischemia-reperfusion model was established after a 7-day gavage intervention, and pathological changes and renal function were observed, using an enzyme-linked immunosorbent assay （ELISA） to detect interleukin 8 （IL-8） and interferon gamma （IFN-r） levels, as well as immunohistochemical staining to detect altered levels of TLR2 and TLR4 expression in renal tissue. Results： After 24 h, renal pathological damage and the expression levels of serum creatinine （Scr）, IL-8, IFN- r, TLR2, and TLR4 were significantly higher in the model group as compared with the pseudo-operation group （P〈0.05）. In addition, at 24 h the above indicators decreased significantly in the Astragaloside group, high- dose SHP group and low-dose SHP group as compared with the ischemia-reperfusion model group （P〈0.05）. TLR2 and TLR4 expression levels were significantly reduced in the SHP treatment and Astragaloside group as compared with the pseudo-operation group （P〈0.05）. Further, the high-dose SHP group showed significantly less renal damage score and decreased levels of TLR expression than those of low-dose SHP group and Astragaloside group （all P〈0.05）. Conclusion： SHP can alleviate the renal structural and functional damage caused by IRI-induced AKI in rats by reducing the damage of renal pathology, which may reduce inflammatory cytokine levels by downregulating the expression of TLRs in renal tissue in a dose-dependent manner.
文摘肾脏缺血时,坏死和凋亡的肾小管上皮及血管内皮细胞会释放出高迁移率族蛋白(high mobility group box 1,HMGB1)。HMGB1可能是诱导肾脏缺血再灌注损伤(renal ischemia and reperfusion injury,RIRI)早期的内源性危险信号。本研究主要阐述了在RIRI早期胞外HMGB1的来源以及与RIRI的关系。HMGB1可促使天然调节型T细胞(nTreg)丧失炎症抑制作用甚至可能翻转为促炎性细胞,该分子同时诱导巨噬细胞分泌大量炎性因子,并作用于活化的巨噬细胞使其自分泌HMGB1,二者共同作用造成肾脏缺血再灌注早期炎症损伤。如在损伤早期阻断HMGB1的多重作用,不仅可以消除巨噬细胞的炎症反应,而且可能恢复nTreg的炎症抑制作用从而减轻炎症反应,可能会对肾脏起到更好的保护作用。
文摘Ischemic postconditioning was defined as rapid intermittent interruptions of blood flow in the early phase of reperfusion,which has been found to be protective against renal ischemia-reperfusion injury(IRI)in animal models but not in clinical trials.We describe a case that the allograft renal vein was twisted because of the surgeon’s mistake,which caused the warm ischemia of allograft after reperfusion.The allograft restored blood flow without second reperfusion and cold preservation after 9 min of warm ischemia.The patient was followed up for 3 months and the allograft worked well without complications.
