反复肢体缺血预处理对大鼠脑缺血再灌注组织NF-κB信号通路及炎症因子含量的影响
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摘要
目的观察肢体反复缺血预处理(limb ischemic preconditioning,LIP)对大鼠脑缺血再灌注损伤后脑组织中核转录因子NF-κB信号通路及肿瘤坏死因子-α(TNF-α)和白介素-6(IL-6)含量的影响,探讨LIP诱导的神经保护机制。方法 84只雄性SD大鼠随机分为3组;假手术组(sham组) 12只,局灶性脑缺血再灌注组(I/R) 36只,预处理+缺血再灌注组(LIP+I/R) 36只;根据再灌注时间不同,后两组分别又分为3个亚组(1,3,7 d),每组12只。分别于脑缺血再灌注后1,3,7 d对各组大鼠进行神经功能损伤评分,HE染色光镜下观察脑组织细胞形态,氯化三苯基四氮唑(TTC)染色方法测定脑梗死体积,免疫组化法测定脑组织中NF-κB/P56的表达,ELISA法检测脑组织中TNF-α、IL-6的含量测定。结果 sham组大鼠无脑梗死病灶及神经功能缺损症状,I/R与LIP+I/R组大鼠脑梗死病灶主要集中在基底节区及额颞顶皮层。与I/R组比较,LIP+I/R组大鼠神经缺损评分降低(P <0.05),脑组织细胞损伤程度减轻(P <0.05),脑梗死体积减小(P <0.05),NF-κB/P56的表达明显下降(P <0.05),TNF-α、IL-6含量明显降低(P <0.05),差异均有统计学意义。结论 LIP可以抑制NF-κB信号通路、降低TNF-α、IL-6的含量来实现内源性保护作用,这可能与减轻炎症反应有关。
Objective To observe the effects of limb ischemic preconditioning(LIP) on the NF-κB signaling pathway,tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels in rats with cerebral ischemia reperfusion injury,and explore the mechanism of LIPinduced neuroprotection. Methods A total of 84 male SD rats were randomly divided into three groups: sham operation group(n =12),cerebral ischemia reperfusion(I/R) group(n = 36),and LIP + I/R group(n = 36). The rats in I/R group and LIP + I/R group were divided into three subgroups(12 in each subgroup) according to different reperfusion time(1,3,7 d),respectively. The neurological injury scores were used to measure the degree of neurological impairment,the cell morphology of brain tissue was observed under the light microscope with HE staining,the volume of cerebral infarction was determined by TTC staining,the expression of NF-κB/P56 in brain tissue was determined by immunohistochemistry,and the contents of TNF-α and IL-6 in brain tissue were determined by ELISA at 1,3,7 d after cerebral ischemia and reperfusion. Results In sham group,rats had no symptoms of cerebral infarction lesion and nerve function injury. The cerebral infarction lesions in I/R group and LIP + I/R group were mainly concentrated in the basal ganglia and frontotemporal parietal cortex. Compared with I/R group,the neurological injury scores,the degree of brain tissue damage,the size of cerebral infarction,the expression of NF-κB/P56 and the contents of TNF-α and IL-6 were decreased significantly in LIP + I/R group(P < 0.05). Conclusion LIP can have endogenous protective effect by inhibiting the NF-κB signaling pathway,decreasing the content of TNF-α and IL-6,and reducing the inflammatory response.
Objective To observe the effects of limb ischemic preconditioning(LIP) on the NF-κB signaling pathway,tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) levels in rats with cerebral ischemia reperfusion injury,and explore the mechanism of LIPinduced neuroprotection. Methods A total of 84 male SD rats were randomly divided into three groups: sham operation group(n =12),cerebral ischemia reperfusion(I/R) group(n = 36),and LIP + I/R group(n = 36). The rats in I/R group and LIP + I/R group were divided into three subgroups(12 in each subgroup) according to different reperfusion time(1,3,7 d),respectively. The neurological injury scores were used to measure the degree of neurological impairment,the cell morphology of brain tissue was observed under the light microscope with HE staining,the volume of cerebral infarction was determined by TTC staining,the expression of NF-κB/P56 in brain tissue was determined by immunohistochemistry,and the contents of TNF-α and IL-6 in brain tissue were determined by ELISA at 1,3,7 d after cerebral ischemia and reperfusion. Results In sham group,rats had no symptoms of cerebral infarction lesion and nerve function injury. The cerebral infarction lesions in I/R group and LIP + I/R group were mainly concentrated in the basal ganglia and frontotemporal parietal cortex. Compared with I/R group,the neurological injury scores,the degree of brain tissue damage,the size of cerebral infarction,the expression of NF-κB/P56 and the contents of TNF-α and IL-6 were decreased significantly in LIP + I/R group(P < 0.05). Conclusion LIP can have endogenous protective effect by inhibiting the NF-κB signaling pathway,decreasing the content of TNF-α and IL-6,and reducing the inflammatory response.
引文
[1]Kitagawa K,Matsumoto M,Tagaya M,et al.Ischemic tolerance phenomenon found in the brain[J].Brain Res,1990,528(1):21-24.
[2]Abdul-Ghani S,Fleishman AN,Khaliulin I,et al.Remote ischemic preconditioning triggers changes in autonomic nervous system activity:implications for cardioprotection[J].Physiol Rep,2017,5(3):1-13.
[3]Zhang Y,Zhang X,Chi D,et al.Remote ischemic preconditioning for prevention of acute kidney injury in patients undergoing on pump cardiac surgery:a systematic review and meta-analysis[J].Medicine(Baltimore),2016,95(37):1465-1476.
[4]He N,Jia JJ,Li JH,et al.Remote ischemic perconditioning prevents liver transplantation induced ischemia/reperfusion injury in rats:role of ROS/RNS and e NOS[J].World J Gastroenterol,2017,23(5):830-841.
[5]Zhao HG,Sun XC,Xian XH,et al.The role of nitric oxide in the neuroprotection of limb ischemic preconditioning in rats[J].Neurochem Res,2007,32(11):1919-1926.
[6]Park UJ,Kim HT,Cho WH,et al.Remote ischemic preconditioning enhances the expression of genes encoding antioxidant enzymes and endoplasmic reticulum stress-related proteins in pat skeletal muscle[J].Vasc Specialist Int,2016,32(4):141-149.
[7]Mudaliar H,Rayner B,Billah M,et al.Remote ischemic preconditioning attenuates EGR-1 expression following myocardial ischemia reperfusion injury through activation of the JAK-STATpathway[J].Inter J Cardiol,2017,228:729-741.
[8]吴江,贾建平.神经病学[M].3版.北京:人民卫生出版社,2015:180.
[9]Chen Y,Zhang L,Ni J,et al.Lldt-8 protects against cerebral ischemia/reperfusion injury by suppressing post-stroke inflammation[J].J Pharmacol Sci,2016,131(2):131-137.
[10]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[11]Ren C,Wang P,Wang B,et al.Limb remote ischemic per-conditioning in combination with post-conditioning reduces brain damage and promotes neuroglobin expression in the rat brain after ischemic stroke[J].Restor Neurol Neurosci,2015,33(3):369-379.
[12]Pavlova S,Klucska K,Vasicek D,et al.The involvement of SIRT1and transcription factor NF-κB(p50/p65)in regulation of porcine ovarian cell function[J].Anim Reprod Sci,2013,140(3-4):180-188.
[13]Calzaseia T,Pellegrini M,Hall H,et al.TNF-alpha is critical for antitumor but not antiviral T cell immunity in mice[J].J Clin Invest,2007,117(12):3833-3845.
[14]Williams AJ,Hale SL,Moffett JR,et al.Delayed treatment with MLN519 reduces infarction and associated neurologic deficit caused by focal ischemic brain injury in rats via antiinflammatory mechanisms involving nuclear factor-KappaB activation,gliosis,and leukocyte infiltration[J].J Cereb Blood Flow Metab,2003,23(1):75-87.
[2]Abdul-Ghani S,Fleishman AN,Khaliulin I,et al.Remote ischemic preconditioning triggers changes in autonomic nervous system activity:implications for cardioprotection[J].Physiol Rep,2017,5(3):1-13.
[3]Zhang Y,Zhang X,Chi D,et al.Remote ischemic preconditioning for prevention of acute kidney injury in patients undergoing on pump cardiac surgery:a systematic review and meta-analysis[J].Medicine(Baltimore),2016,95(37):1465-1476.
[4]He N,Jia JJ,Li JH,et al.Remote ischemic perconditioning prevents liver transplantation induced ischemia/reperfusion injury in rats:role of ROS/RNS and e NOS[J].World J Gastroenterol,2017,23(5):830-841.
[5]Zhao HG,Sun XC,Xian XH,et al.The role of nitric oxide in the neuroprotection of limb ischemic preconditioning in rats[J].Neurochem Res,2007,32(11):1919-1926.
[6]Park UJ,Kim HT,Cho WH,et al.Remote ischemic preconditioning enhances the expression of genes encoding antioxidant enzymes and endoplasmic reticulum stress-related proteins in pat skeletal muscle[J].Vasc Specialist Int,2016,32(4):141-149.
[7]Mudaliar H,Rayner B,Billah M,et al.Remote ischemic preconditioning attenuates EGR-1 expression following myocardial ischemia reperfusion injury through activation of the JAK-STATpathway[J].Inter J Cardiol,2017,228:729-741.
[8]吴江,贾建平.神经病学[M].3版.北京:人民卫生出版社,2015:180.
[9]Chen Y,Zhang L,Ni J,et al.Lldt-8 protects against cerebral ischemia/reperfusion injury by suppressing post-stroke inflammation[J].J Pharmacol Sci,2016,131(2):131-137.
[10]Longa EZ,Weinstein PR,Carlson S,et al.Reversible middle cerebral artery occlusion without craniectomy in rats[J].Stroke,1989,20(1):84-91.
[11]Ren C,Wang P,Wang B,et al.Limb remote ischemic per-conditioning in combination with post-conditioning reduces brain damage and promotes neuroglobin expression in the rat brain after ischemic stroke[J].Restor Neurol Neurosci,2015,33(3):369-379.
[12]Pavlova S,Klucska K,Vasicek D,et al.The involvement of SIRT1and transcription factor NF-κB(p50/p65)in regulation of porcine ovarian cell function[J].Anim Reprod Sci,2013,140(3-4):180-188.
[13]Calzaseia T,Pellegrini M,Hall H,et al.TNF-alpha is critical for antitumor but not antiviral T cell immunity in mice[J].J Clin Invest,2007,117(12):3833-3845.
[14]Williams AJ,Hale SL,Moffett JR,et al.Delayed treatment with MLN519 reduces infarction and associated neurologic deficit caused by focal ischemic brain injury in rats via antiinflammatory mechanisms involving nuclear factor-KappaB activation,gliosis,and leukocyte infiltration[J].J Cereb Blood Flow Metab,2003,23(1):75-87.