红景天苷预处理对大鼠移植肾缺血再灌注损伤的保护
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摘要
目的探讨红景天苷预处理对移植肾缺血再灌注损伤的保护机制。方法建立近交系雄性SD大鼠同基因肾移植模型。共设3个组,假手术组、红景天苷预处理组、移植缺血再灌注模型组,每组24只。观察红景天苷预处理对大鼠移植肾缺血再灌注过程中血清尿素氮(BUN)、肌酐(SCr)、过氧化物歧化酶(SOD)、脂质过氧化产物丙二醛(MDA)水平变化的影响;取移植肾组织HE染色后光镜观察肾组织病理学改变;荧光探针DCFH-DA检测活性氧(ROS)含量;采用免疫组织化学染色和放射免疫法分别检测肾组织中核转录因子κB(NF-κB)阳性细胞数目和肿瘤坏死因子-α(TNF-α)、白介素1(IL-1)、白介素-6(IL-6)的表达。结果红景天苷预处理组大鼠BUN、SCr、MDA在术后轻度升高,与假手术组比较无统计学差异(P>0.05),与模型组比较有统计学差异(P<0.01);SOD在术后24 h增加最明显,与假手术组比较有统计学差异(P<0.05),与模型组比较有统计学差异(P<0.05)。ROS含量在红景天苷预处理组术后上升,与假手术组比较有统计学差异(P<0.05),但不如模型组升高明显,两组比较有统计学差异(P<0.01)。红景天苷预处理组肾脏病理表现损伤较轻,与模型组比较有统计学差异(P<0.05)。NF-κB阳性细胞数目在红景天苷预处理组术后表达略增加,在模型组表达明显增加,两组比较有统计学差异(P<0.01)。肾组织匀浆中TNF-α、IL-1和IL-6含量在假手术组和红景天苷预处理组各时间段均未见明显变化,两组间比较无统计学差异(P>0.05),而模型组含量明显增加,与其他两组比较差异有显著性(P<0.01)。结论红景天苷预处理通过抑制氧化应激和减轻炎性因子对大鼠肾脏移植缺血再灌注后损伤具有一定的预防和保护作用。
Objective To explore the mechanism of prevention of salidroside pre-treatment against ischemia-reperfusion injury in rats after kidney transplantation.Methods The allogeneic kidney transplantation model inbred male SD(Sprague Dawley,SD) rats was established.All of rats were randomly divided into 3 groups(24 one in each group): sham-operation group,salidroside pretreatment group and ischemia-reperfusion transplantation model group.Effects of salidroside pre-treatment on the levels of blood urea nitrogen(BUN),serum creatinine(SCr),superoxide dismutase(SOD) and malondialdehyde(MDA) during ischemia-reperfusion in the transplanted kidney were observed in the rats.The transplanted kidney tissue was taken to conduct HE staining,and then pathological changes in the kidney tissue were observed under an optical microscope.Content of reactive oxygen species(ROS) was detected with the fluorescent probe DCFH-DA.Immunohistochemical staining and immunoradiometric assay method were adopted to detect the number of nuclear transcription factor κB(NF-κB) positive cells and expression of tumor necrosis factor-α(TNF-α),interleukin 1(IL-1) and interleukin 6(IL-6) in the kidney tissue.Results After operation,BUN,SCr and MDA in the rats in the salidroside pretreatment group increased slightly;and,compared to the sham-operation group there was no statistical difference(P>0.05),compared to the model group there was statistical difference(P<0.01).SOD values in the salidroside pretreatment group were increased most significantly at 24 h after operation;and,compared to the sham-operation group there was statistical difference(P<0.05),and compared to the model group there was statistical difference(P<0.05).Content of ROS in the salidroside pretreatment group increased after operation;and,compared to the sham-operation group there was statistical difference(P<0.05).But the increase was not as significant as that in the model group,and there was statistical difference between the two groups(P<0.01).Renal pathology in the salidroside pretreatment group showed less injury,and compared to the model group there was statistical difference(P<0.05).Number of NF-κB positive cell in the salidroside pretreatment group showed slight increase and significant increase in the model group,and there was statistical difference between the two groups(P<0.01).Contents of TNF-a,IL-1 and IL-6 in the homogenate of kidney tissues had not any significant changes at each time period in both the sham-operation group and the salidroside pretreatment group,with no statistical difference between the two groups salidroside pretreatment group;while in the model group the contents increased significantly,with statistical significance of difference compared to the other two groups.Conclusions Salidroside injection pretreatment can prevent and protect against ischemia-reperfusion injury in rats after kidney transplantation by inhibiting oxidative stress reactions and inflammatory cascade reactions.
Objective To explore the mechanism of prevention of salidroside pre-treatment against ischemia-reperfusion injury in rats after kidney transplantation.Methods The allogeneic kidney transplantation model inbred male SD(Sprague Dawley,SD) rats was established.All of rats were randomly divided into 3 groups(24 one in each group): sham-operation group,salidroside pretreatment group and ischemia-reperfusion transplantation model group.Effects of salidroside pre-treatment on the levels of blood urea nitrogen(BUN),serum creatinine(SCr),superoxide dismutase(SOD) and malondialdehyde(MDA) during ischemia-reperfusion in the transplanted kidney were observed in the rats.The transplanted kidney tissue was taken to conduct HE staining,and then pathological changes in the kidney tissue were observed under an optical microscope.Content of reactive oxygen species(ROS) was detected with the fluorescent probe DCFH-DA.Immunohistochemical staining and immunoradiometric assay method were adopted to detect the number of nuclear transcription factor κB(NF-κB) positive cells and expression of tumor necrosis factor-α(TNF-α),interleukin 1(IL-1) and interleukin 6(IL-6) in the kidney tissue.Results After operation,BUN,SCr and MDA in the rats in the salidroside pretreatment group increased slightly;and,compared to the sham-operation group there was no statistical difference(P>0.05),compared to the model group there was statistical difference(P<0.01).SOD values in the salidroside pretreatment group were increased most significantly at 24 h after operation;and,compared to the sham-operation group there was statistical difference(P<0.05),and compared to the model group there was statistical difference(P<0.05).Content of ROS in the salidroside pretreatment group increased after operation;and,compared to the sham-operation group there was statistical difference(P<0.05).But the increase was not as significant as that in the model group,and there was statistical difference between the two groups(P<0.01).Renal pathology in the salidroside pretreatment group showed less injury,and compared to the model group there was statistical difference(P<0.05).Number of NF-κB positive cell in the salidroside pretreatment group showed slight increase and significant increase in the model group,and there was statistical difference between the two groups(P<0.01).Contents of TNF-a,IL-1 and IL-6 in the homogenate of kidney tissues had not any significant changes at each time period in both the sham-operation group and the salidroside pretreatment group,with no statistical difference between the two groups salidroside pretreatment group;while in the model group the contents increased significantly,with statistical significance of difference compared to the other two groups.Conclusions Salidroside injection pretreatment can prevent and protect against ischemia-reperfusion injury in rats after kidney transplantation by inhibiting oxidative stress reactions and inflammatory cascade reactions.
引文
[1] Ramessur Chandran S, Han Y, Tesch GH, et al. Inhibition of spleen tyrosine kinase reduces renal allograft injury in a rat model of acute antibody-mediated rejection in sensitized recipients[J]. Transplantation, 2017, 101(8): e240-e248.
[2] Bao DS, Wu YK, Fu SJ, et al. Hyperbaric oxygenation protects against ischemia-reperfusion injury in transplanted rat kidneys by triggering autophagy and inhibiting inflammatory response[J]. Ann Transplant, 2017, 2(10): 22, 75-82.
[3] 李剑, 范维琥, 罗心平, 等.预灌红景天对心梗大鼠心功能及Flk-lmRNA表达的影响[J]. 复旦学报(医学版),2006,33(3): 384-387.
[4] 赵正维, 王为忠, 陈宏, 等.红景天对大鼠小肠缺血再灌注损伤的保护作用[J].中国临床康复,2006,10(43):117-119.
[5] 隋汝波, 李熙东, 刘媛媛, 等.大鼠局灶性脑缺血再灌注血浆细胞因子和内皮素对红景天干预的反应[J]. 中国临床康复,2006,10(35):23-25.
[6] Lautraite S, Bigot-Lasserre D, Bars R, et al. Optimisation of cell-based assays for medium throughput screening of oxidative stress[J]. Toxicol In Vitro, 2003, 17(2): 207-220.
[7] Malek M, Nematbakhsh M, Renal ischemia/reperfusion injury: from pathophysiology to treatment[J]. J Renal Inj Prev, 2015, 4(2): 20-27.
[8] Alberú-Gómez JI, Hernández-Méndez EA, Oropeza-Barrera I, et al. Incidence of acute rejection in patients with renal graft dysfunction[J]. Rev Invest Clin, 2013, 65(5): 412-419.
[9] Tsuda H, Kawada N, Kaimori JY, et al. Febuxostat suppressed renal ischemia-reperfusion injury via reduced oxidative stress[J]. Biochem Biophys Res Commun, 2012, 427(2): 266-272.
[10] Cao CC, Ding XQ, Ou ZL, et al. In vivo transfection of NF-kappa B decoy oligodeoxynucleotides attenuate renal ischem, i a/reperfusion injury in rats[J]. Kidney Int, 2004,65(3): 834-845.
[11] Yuan Y, Wu SJ, Liu X, et al. Antioxidant effect of salidroside and its protective effect against furan-induced hepatocyte damage in mice[J].Food Funct,2013, 4(5):763-769.
[12] Han T. Effects of salidroside pretreatment on expression of tumor necrosis factor-alpha and permeability of blood brain barrier in rat model of focal cerebral ischemia-reperfusion injury[J]. Asian Pac J Trop Med, 2013, 6(2): 156-158.
[13] Li D, Fu Y, Zhang W, et al. Salidroside attenuates inflammatory responses by suppressing nucleal factor-κB and mitogen activated protein kinases activation in lipopolysaccharide-induced mastitis in mice[J]. Inflammation Res, 2012, 62(2): 9-15.
[14] 邹毅清, 蔡志扬, 李小宝, 等. 红景天苷预处理对大鼠全脑缺血再灌注后炎症反应的影响[J]. 现代中西医结合杂志, 2013, 22(3): 253-255.
[15] Slegtenhorst BR, Dor FJ, Rodrriguez H, et al. Ischemia/reperfusion injury and its consequences on immunity and inflammation[J]. Curr transplant REP, 2014, 1(3): 147-154.
[16] Guan S, Feng H, Song B, et al. Salidroside attenuates LPS-induced pro-inflammatory cytokine responses and improves survival in murine endotoxemia[J]. Int Immunopharmacol, 2011, 11(12): 2194-2199.
[2] Bao DS, Wu YK, Fu SJ, et al. Hyperbaric oxygenation protects against ischemia-reperfusion injury in transplanted rat kidneys by triggering autophagy and inhibiting inflammatory response[J]. Ann Transplant, 2017, 2(10): 22, 75-82.
[3] 李剑, 范维琥, 罗心平, 等.预灌红景天对心梗大鼠心功能及Flk-lmRNA表达的影响[J]. 复旦学报(医学版),2006,33(3): 384-387.
[4] 赵正维, 王为忠, 陈宏, 等.红景天对大鼠小肠缺血再灌注损伤的保护作用[J].中国临床康复,2006,10(43):117-119.
[5] 隋汝波, 李熙东, 刘媛媛, 等.大鼠局灶性脑缺血再灌注血浆细胞因子和内皮素对红景天干预的反应[J]. 中国临床康复,2006,10(35):23-25.
[6] Lautraite S, Bigot-Lasserre D, Bars R, et al. Optimisation of cell-based assays for medium throughput screening of oxidative stress[J]. Toxicol In Vitro, 2003, 17(2): 207-220.
[7] Malek M, Nematbakhsh M, Renal ischemia/reperfusion injury: from pathophysiology to treatment[J]. J Renal Inj Prev, 2015, 4(2): 20-27.
[8] Alberú-Gómez JI, Hernández-Méndez EA, Oropeza-Barrera I, et al. Incidence of acute rejection in patients with renal graft dysfunction[J]. Rev Invest Clin, 2013, 65(5): 412-419.
[9] Tsuda H, Kawada N, Kaimori JY, et al. Febuxostat suppressed renal ischemia-reperfusion injury via reduced oxidative stress[J]. Biochem Biophys Res Commun, 2012, 427(2): 266-272.
[10] Cao CC, Ding XQ, Ou ZL, et al. In vivo transfection of NF-kappa B decoy oligodeoxynucleotides attenuate renal ischem, i a/reperfusion injury in rats[J]. Kidney Int, 2004,65(3): 834-845.
[11] Yuan Y, Wu SJ, Liu X, et al. Antioxidant effect of salidroside and its protective effect against furan-induced hepatocyte damage in mice[J].Food Funct,2013, 4(5):763-769.
[12] Han T. Effects of salidroside pretreatment on expression of tumor necrosis factor-alpha and permeability of blood brain barrier in rat model of focal cerebral ischemia-reperfusion injury[J]. Asian Pac J Trop Med, 2013, 6(2): 156-158.
[13] Li D, Fu Y, Zhang W, et al. Salidroside attenuates inflammatory responses by suppressing nucleal factor-κB and mitogen activated protein kinases activation in lipopolysaccharide-induced mastitis in mice[J]. Inflammation Res, 2012, 62(2): 9-15.
[14] 邹毅清, 蔡志扬, 李小宝, 等. 红景天苷预处理对大鼠全脑缺血再灌注后炎症反应的影响[J]. 现代中西医结合杂志, 2013, 22(3): 253-255.
[15] Slegtenhorst BR, Dor FJ, Rodrriguez H, et al. Ischemia/reperfusion injury and its consequences on immunity and inflammation[J]. Curr transplant REP, 2014, 1(3): 147-154.
[16] Guan S, Feng H, Song B, et al. Salidroside attenuates LPS-induced pro-inflammatory cytokine responses and improves survival in murine endotoxemia[J]. Int Immunopharmacol, 2011, 11(12): 2194-2199.