中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2015年
1期
66-70
,共5页
陶晓刚%刘佰运%陈学涛%郝淑煜%田润发
陶曉剛%劉佰運%陳學濤%郝淑煜%田潤髮
도효강%류백운%진학도%학숙욱%전윤발
颅脑损伤%聚腺苷二磷酸核糖聚合酶%NF-κB%炎性反应%小鼠
顱腦損傷%聚腺苷二燐痠覈糖聚閤酶%NF-κB%炎性反應%小鼠
로뇌손상%취선감이린산핵당취합매%NF-κB%염성반응%소서
Craniocerebral trauma%Poly (ADP-ribose) polymerase%Nuclear factor-κB%Inflammatory response%Mice
目的 探讨聚腺苷二磷酸核糖聚合酶(PARP)抑制剂PJ34对小鼠创伤性颅脑损伤(TBI)的保护作用.方法 将BALB/c雄性小鼠分为假手术组、模型组和PJ34组,建立小鼠可控性皮质打击伤(CCI)模型.分别在打击后6h、24h从运动、感觉、反射和平衡等方面评价小鼠的神经功能缺损程度,并测定脑挫裂伤体积.打击后24h时通过HE染色评价受损脑区神经元的损伤.分别在打击后6h、24h采用髓过氧化物酶(MPO)活性测定试剂盒测定损伤脑组织内MPO的活性;并采用Western blot方法分别检测细胞质和细胞核NF-κB p65亚单位的含量以及细胞质内炎性反应因子TNF-α和IL-1β的含量.结果 PJ34组打击后6h、24h神经功能评分分别为(8.20 ±0.25)分和(7.80±0.25)分,均低于模型组(12.40-±0.38)分和(11.70±0.22)分(均P＜0.01);脑挫裂伤体积分别为(10.25 ±0.61) mm3和(11.55±1.16) mm3,均小于模型组(25.07±1.45) mm3和(27.24±1.51) mm3(均P＜0.01);MPO活性分别为(0.013 ±0.001) U/g和(0.018-±0.001) U/g,均低于模型组(0.024 ±0.001) U/g和(0.023 ±0.001) U/g(均P ＜0.05);细胞质内NF-κB的表达水平均低于模型组(均P＜0.05).PJ34组打击后24h细胞核内NF-κB的表达水平低于模型组(P＜0.01),而6h细胞核内NF-κB水平与模型组之间的差异无统计学意义.PJ34组打击后6h和24h的TNF-α及IL-1β的表达水平均低于模型组,差异均有统计学意义(P＜0.05或P＜0.01).结论 PARP-NF-κB炎性反应通路在TBI后的继发性损伤中起重要作用,PJ34可以阻断该通路,抑制TBI后的炎性反应,从而起到脑保护的作用.
目的 探討聚腺苷二燐痠覈糖聚閤酶(PARP)抑製劑PJ34對小鼠創傷性顱腦損傷(TBI)的保護作用.方法 將BALB/c雄性小鼠分為假手術組、模型組和PJ34組,建立小鼠可控性皮質打擊傷(CCI)模型.分彆在打擊後6h、24h從運動、感覺、反射和平衡等方麵評價小鼠的神經功能缺損程度,併測定腦挫裂傷體積.打擊後24h時通過HE染色評價受損腦區神經元的損傷.分彆在打擊後6h、24h採用髓過氧化物酶(MPO)活性測定試劑盒測定損傷腦組織內MPO的活性;併採用Western blot方法分彆檢測細胞質和細胞覈NF-κB p65亞單位的含量以及細胞質內炎性反應因子TNF-α和IL-1β的含量.結果 PJ34組打擊後6h、24h神經功能評分分彆為(8.20 ±0.25)分和(7.80±0.25)分,均低于模型組(12.40-±0.38)分和(11.70±0.22)分(均P＜0.01);腦挫裂傷體積分彆為(10.25 ±0.61) mm3和(11.55±1.16) mm3,均小于模型組(25.07±1.45) mm3和(27.24±1.51) mm3(均P＜0.01);MPO活性分彆為(0.013 ±0.001) U/g和(0.018-±0.001) U/g,均低于模型組(0.024 ±0.001) U/g和(0.023 ±0.001) U/g(均P ＜0.05);細胞質內NF-κB的錶達水平均低于模型組(均P＜0.05).PJ34組打擊後24h細胞覈內NF-κB的錶達水平低于模型組(P＜0.01),而6h細胞覈內NF-κB水平與模型組之間的差異無統計學意義.PJ34組打擊後6h和24h的TNF-α及IL-1β的錶達水平均低于模型組,差異均有統計學意義(P＜0.05或P＜0.01).結論 PARP-NF-κB炎性反應通路在TBI後的繼髮性損傷中起重要作用,PJ34可以阻斷該通路,抑製TBI後的炎性反應,從而起到腦保護的作用.
목적 탐토취선감이린산핵당취합매(PARP)억제제PJ34대소서창상성로뇌손상(TBI)적보호작용.방법 장BALB/c웅성소서분위가수술조、모형조화PJ34조,건립소서가공성피질타격상(CCI)모형.분별재타격후6h、24h종운동、감각、반사화평형등방면평개소서적신경공능결손정도,병측정뇌좌렬상체적.타격후24h시통과HE염색평개수손뇌구신경원적손상.분별재타격후6h、24h채용수과양화물매(MPO)활성측정시제합측정손상뇌조직내MPO적활성;병채용Western blot방법분별검측세포질화세포핵NF-κB p65아단위적함량이급세포질내염성반응인자TNF-α화IL-1β적함량.결과 PJ34조타격후6h、24h신경공능평분분별위(8.20 ±0.25)분화(7.80±0.25)분,균저우모형조(12.40-±0.38)분화(11.70±0.22)분(균P＜0.01);뇌좌렬상체적분별위(10.25 ±0.61) mm3화(11.55±1.16) mm3,균소우모형조(25.07±1.45) mm3화(27.24±1.51) mm3(균P＜0.01);MPO활성분별위(0.013 ±0.001) U/g화(0.018-±0.001) U/g,균저우모형조(0.024 ±0.001) U/g화(0.023 ±0.001) U/g(균P ＜0.05);세포질내NF-κB적표체수평균저우모형조(균P＜0.05).PJ34조타격후24h세포핵내NF-κB적표체수평저우모형조(P＜0.01),이6h세포핵내NF-κB수평여모형조지간적차이무통계학의의.PJ34조타격후6h화24h적TNF-α급IL-1β적표체수평균저우모형조,차이균유통계학의의(P＜0.05혹P＜0.01).결론 PARP-NF-κB염성반응통로재TBI후적계발성손상중기중요작용,PJ34가이조단해통로,억제TBI후적염성반응,종이기도뇌보호적작용.
Objective To investigate whether PARP inhibitor,PJ34,participates in the inflammation related to PARP-NF-κB pathway in a mouse model of controlled cortical impact (CCI).Methods One hundred and thirty-six BALB/c male mice were divided into 3 groups:sham-operated group,vehicle-treated group and PJ34-treated group.Injury to the cerebral cortex was produced with controlled cortical impact.Evaluation of the neurological deficits was performed at 6 h and 24 h after CCI,which included the motor,sensory,reflex testing and beam balance testing.The contusion volumes were measured after HE staining at 6 h and 24 h after CCI.The activities of MPO in contusion cortex were examined by using MPO ELISA kit.Western blot was performed to detect the levels of NF-κB p65 in cytosolic and nuclear fractions and to detect TNF-α and IL-1β in cytosolic fractions.Results Neurological severity scores at 6 h and 24 h after CCI in PJ34-treated group(8.20 ±0.25 and 7.80 ±0.25,respectively) are less than those in vehicle-treated group(12.40 ±0.38 and 11.70 ±0.22,respectively) (all P ＜0.01).The contusion volume at 6 h and 24 h after CCI in PJ34-treated group[(10.25 ± 0.61) mm3 and (11.55 ±1.16) mm3,respectively] is less than that in vehicle-treated group[(25.07 ± 1.45)mm3 and (27.24 ± 1.51)mm3,respectively] (all P ＜ 0.01).The activities of MPO at 6 h and 24 h after CCI in PJ34-treated group [(0.013 ±0.001) U/g and (0.018 ± 0.001) U/g,respectively] is less than those in vehicle-treated group [(0.024 ± 0.001) U/g and (0.023 ± 0.001) U/g,respectively] (all P ＜ 0.05).Moreover,the expression level of NF-κB p65 in cytoplasm at 6 h and 24 h after CCI in PJ34-treated group is less than that in vehicle-treated group (all P ＜ 0.05).Compared with vehicle-treated group,the expression level of NF-κB p65 in cell nuclei at 24 h after CCI in PJ34-treated group is higher (P ＜0.01).There is no significant difference between the expression level of NF-κB p65 in cell nuclei at 6 h after CCI in PJ34-treated group and in vehicle-treated group.PJ34 also reduced the expression level of TNF-α and IL-13 at 6 h or 24 h after CCI(P ＜ 0.05 or P ＜ 0.01).Conclusions PARP-NF-κB inflammatory pathway plays an important role during the secondary injury after traumatic brain injury(TBI).PJ34 could block the pathway,inhibit inflammatory response and protect the brain against TBI.
