ω-3多不饱和脂肪酸对大鼠创伤性脑损伤后神经功能的影响
ω-3다불포화지방산대대서창상성뇌손상후신경공능적영향
Effect of omega-3 polyunsaturated fatty acids supplementation on traumatic brain injury in a rat model
  • 万方数据
    中华临床营养杂志 중화림상영양잡지
    CHINESE JOURNAL OF CLINICAL NUTRITION
    2015年 2期 110-115 ,共6页

    陈祥荣%谢宝缘%骆良钦%李亚松%曾以勒

    진상영%사보연%락량흠%리아송%증이륵

    创伤性脑损伤%ω-3多不饱和脂肪酸%自噬%神经保护 創傷性腦損傷%ω-3多不飽和脂肪痠%自噬%神經保護
    창상성뇌손상%ω-3다불포화지방산%자서%신경보호
    Traumatic brain injury%Omega-3 polyunsaturated fatty acids%Autophagy%Neuroprotection
    目的 探讨ω-3多不饱和脂肪酸(ω-3 PUFA)对大鼠创伤性脑损伤(TBI)后脑水肿、自噬反应及神经功能的影响及其机制.方法 采用改良Feeney法建立大鼠TBI模型,将72只SD大鼠采用随机数字表法分为假手术组、TBI组、TBI+ ω-3 PUFA处理组(TBI+ ω-3组)和TBI+自噬抑制剂3-甲基腺嘌呤组(TBI+3-MA组)(n均=18),各组再根据3个时间点(创伤后第1、3、7天)分设3个亚组(n=6).于不同时间点采用改良神经功能评分表(mNSS)进行神经功能评分,采用干湿重法测量损伤区脑组织水含量,于建模后第3天采用免疫荧光、PCR和Westerrnblot法测定自噬相关因子(LC3-Ⅱ、Beclin-1)的mRNA及蛋白表达水平.结果 损伤后第1、3、7天,与假手术组比较,TBI组、TBI+ ω-3组和TBI+ 3-MA组的mNSS评分(TBI组:12.42 ±0.27比1.34±0.32,12.07±0.27比1.16±0.29,10.22±0.39比1.22±0.30;TBI+ ω-3组:12.05±0.23比1.34±0.32,11.38±0.21比1.16±0.29,8.20±0.21比1.22±0.30;TBI+ 3-MA组:11.93±0.20比1.34±0.32,11.09±0.19比1.16±0.29,7.93±0.17比1.22±0.30;P均=0.00)和脑水含量均显著升高[TBI组:(79.82±0.61)％比(71.87±0.43)％,(83.04±0.42)％比(72.13±0.53)％,(75.12±0.72)％比(71.78±0.38)％;TBI +ω-3组:(76.81±0.63)％比(71.87±0.43)％,(79.39±0.59)％比(72.13±0.53)％,(73.86±0.38)％比(71.78±0.38)％;TBI+ 3-MA组:(75.98±0.49)％比(71.87±0.43)％,(77.14±0.46)％比(72.13±0.53)％,(72.24±0.37)％比(71.78±0.38)％;P均=0.00].模型建立后第3天,TBI组、TBI+ ω-3组和TBI+ 3-MA组脑组织LC3-Ⅱ和Beclin-1的mRNA及蛋白表达均显著上调(P均=0.00).模型建立后第3、7天,TBI+ ω-3组和TBI+ 3-MA组的mNSS评分均显著低于TBI组(TBI+ ω-3组:11.38±0.21比12.07±0.27、P=0.04,8.20±0.21比10.22±0.39、P=0.01;TBI+ 3-MA组:11.09±0.19比12.07±0.27、P=0.01,7.93±0.17比10.22±0.39、P=0.00);模型建立后第1、3、7天脑组织含水量均显著低于TBI组[TBI+ ω-3组:(76.81±0.63)％比(79.82±0.61)％、P=0.04,(79.39±0.59)％比(83.04±0.42)％、P=0.01,(73.86±0.38)％比(75.12±0.72)％、P=0.03;TBI+ 3-MA组:(75.98±0.49)％比(79.82±0.61)％、P=0.01,(77.14±0.46)％比(83.04±0.42)％、P=0.00,(72.24±0.37)％比(75.12±0.72)％、P=0.02];第3天脑组织LC3-Ⅱ和Beclin-1的mRNA(TBI+ ω-3组:P=0.04,P=0.01;TBI+ 3-MA组:P=0.01,P=0.00)及蛋白表达(TBI+ω-3组:P=0.01,P=0.03;TBI+3-MA组:P均=0.00)均显著低于TBI组.结论 ω-3 PUFA可减轻大鼠TBI后脑水肿,改善创伤后神经功能,具有神经保护作用,其机制可能与降低颅脑损伤后神经细胞自噬反应相关.
    목적 탐토ω-3다불포화지방산(ω-3 PUFA)대대서창상성뇌손상(TBI)후뇌수종、자서반응급신경공능적영향급기궤제.방법 채용개량Feeney법건립대서TBI모형,장72지SD대서채용수궤수자표법분위가수술조、TBI조、TBI+ ω-3 PUFA처리조(TBI+ ω-3조)화TBI+자서억제제3-갑기선표령조(TBI+3-MA조)(n균=18),각조재근거3개시간점(창상후제1、3、7천)분설3개아조(n=6).우불동시간점채용개량신경공능평분표(mNSS)진행신경공능평분,채용간습중법측량손상구뇌조직수함량,우건모후제3천채용면역형광、PCR화Westerrnblot법측정자서상관인자(LC3-Ⅱ、Beclin-1)적mRNA급단백표체수평.결과 손상후제1、3、7천,여가수술조비교,TBI조、TBI+ ω-3조화TBI+ 3-MA조적mNSS평분(TBI조:12.42 ±0.27비1.34±0.32,12.07±0.27비1.16±0.29,10.22±0.39비1.22±0.30;TBI+ ω-3조:12.05±0.23비1.34±0.32,11.38±0.21비1.16±0.29,8.20±0.21비1.22±0.30;TBI+ 3-MA조:11.93±0.20비1.34±0.32,11.09±0.19비1.16±0.29,7.93±0.17비1.22±0.30;P균=0.00)화뇌수함량균현저승고[TBI조:(79.82±0.61)％비(71.87±0.43)％,(83.04±0.42)％비(72.13±0.53)％,(75.12±0.72)％비(71.78±0.38)％;TBI +ω-3조:(76.81±0.63)％비(71.87±0.43)％,(79.39±0.59)％비(72.13±0.53)％,(73.86±0.38)％비(71.78±0.38)％;TBI+ 3-MA조:(75.98±0.49)％비(71.87±0.43)％,(77.14±0.46)％비(72.13±0.53)％,(72.24±0.37)％비(71.78±0.38)％;P균=0.00].모형건립후제3천,TBI조、TBI+ ω-3조화TBI+ 3-MA조뇌조직LC3-Ⅱ화Beclin-1적mRNA급단백표체균현저상조(P균=0.00).모형건립후제3、7천,TBI+ ω-3조화TBI+ 3-MA조적mNSS평분균현저저우TBI조(TBI+ ω-3조:11.38±0.21비12.07±0.27、P=0.04,8.20±0.21비10.22±0.39、P=0.01;TBI+ 3-MA조:11.09±0.19비12.07±0.27、P=0.01,7.93±0.17비10.22±0.39、P=0.00);모형건립후제1、3、7천뇌조직함수량균현저저우TBI조[TBI+ ω-3조:(76.81±0.63)％비(79.82±0.61)％、P=0.04,(79.39±0.59)％비(83.04±0.42)％、P=0.01,(73.86±0.38)％비(75.12±0.72)％、P=0.03;TBI+ 3-MA조:(75.98±0.49)％비(79.82±0.61)％、P=0.01,(77.14±0.46)％비(83.04±0.42)％、P=0.00,(72.24±0.37)％비(75.12±0.72)％、P=0.02];제3천뇌조직LC3-Ⅱ화Beclin-1적mRNA(TBI+ ω-3조:P=0.04,P=0.01;TBI+ 3-MA조:P=0.01,P=0.00)급단백표체(TBI+ω-3조:P=0.01,P=0.03;TBI+3-MA조:P균=0.00)균현저저우TBI조.결론 ω-3 PUFA가감경대서TBI후뇌수종,개선창상후신경공능,구유신경보호작용,기궤제가능여강저로뇌손상후신경세포자서반응상관.
    Objective To investigate the effects of omega-3 polyunsaturated fatty acids (ω-3 PUFA)supplementation on brain edema,autophagy response and neurobehavioral outcome after traumatic brain injury (TBI) in rats and the related mechanisms.Methods TBI rat models were established using Feeney's method.Seventy-two SD rats were divided into 4 groups using random number table:sham operation group,TBI group,ω-3 PUFA supplementation group (TBI + ω-3 group) and autophagy inhibitor 3-methyladenine group (TBI + 3-MA group) (all n =18),each group was further divided into 3 sub-groups (n =6) corresponding to 3 time points (days 1,3,and 7 after TBI).On each of the 3 time points,we measured rat behavioral outcomes with modified neurologic severity score (mNSS) tests;brain water content was measured with wet-dry weight method.The mRNA and protein expressions of autophagy-related factors (LC3-Ⅱ and Beclin-1) in TBI cerebral cortex were determined by immunohistochemistry staining,reverse transcription-polymerase chain reaction and Western blot on day 3 after TBI.Results Compared with the sham group,on days 1,3,and 7 after injuary,the TBI group,the TBI + ω-3 group,and the TBI + 3-MA group had significantly higher mNSS scores (TBI group:12.42±0.27vs.1.34±0.32,12.07±0.27vs.1.16±0.29,10.22±0.39vs.1.22±0.30;TBI+ω-3 group:12.05 ±0.23 vs.1.34 ±0.32,11.38 ±0.21 vs.1.16±0.29,8.20 ±0.21 vs.1.22±0.30;TBI +3-MA group:11.93 ±0.20 vs.1.34 ±0.32,11.09 ±0.19 vs.1.16 ±0.29,7.93 ±0.17 vs.1.22 ± 0.30;all P =0.00) and brain water content [TBI group:(79.82 ± 0.61) ％ vs.(71.87 ± 0.43) ％,(83.04±0.42)％ vs.(72.13 ±0.53)％,(75.12 ±0.72)％ vs.(71.78 ±0.38)％;TBI+ω-3 group:(76.81 ±0.63)％ vs.(71.87 ±0.43)％,(79.39 ±0.59)％ vs.(72.13 ±0.53)％,(73.86 ±0.38)％ vs.(71.78 ±0.38)％;TBI+3-MAgroup:(75.98 ±0.49)％ vs.(71.87 ±0.43)％,(77.14 ±0.46)％ vs.(72.13 ±0.53)％,(72.24 ±0.37)％ vs.(71.78 ±0.38)％;all P =0.00].The mRNA and protein expressions of LC3-Ⅱ and Beclin-1 in the brain were also significantly higher on day 3 in the TBI group,the TBI + ω-3 group,and the TBI + 3-MA group (all P =0.00).Compared with the TB1 group,on day 3 and day 7 after injury,the TBI + ω-3 group and the TBI + 3-MA group had significantly lower mNSS scores (TBI + ω-3 group:11.38±0.21 vs.12.07±0.27,P=0.04,8.20±0.21 vs.10.22±0.39,P=0.01;TBI+3-MA group:11.09±0.19vs.12.07 ± 0.27,P=0.01,7.93 ± 0.17 vs.10.22±0.39,P=0.00).Ondays1,3,and 7,compared with the TBI group,the TBI + ω-3 group and the TBI + 3-MA group had significantly lower brain water content [TBI + ω-3 group:(76.81 ± 0.63) ％ vs.(79.82 ± 0.61) ％,P =0.04,(79.39 ±0.59)％ vs.(83.04±0.42)％,P=0.01,(73.86±0.38)％ vs.(75.12±0.72)％,P=0.03;TBI+3-MAgroup:(75.98 ±0.49)％ vs.(79.82 ±0.61)％,P=0.01,(77.14 ±0.46)％ vs.(83.04 ±0.42)％,P =0.00,(72.24 ± 0.37) ％ vs.(75.12 ± 0.72) ％,P =0.02].On day 3,the TBI + ω-3 group and the TBI + 3-MA group had significantly reduced LC3-Ⅱ and Beclin-1 mRNA expression compared with the TBI group (TBI +ω-3 group:P=0.04,P =0.01;TBI +3-MA group:P =0.01,P =0.00) and protein expression (TBI+ω-3 group:P=0.01,P=0.03;TBI +3-MA group:both P=0.00).Conclusion ω-3 PUFA supplementation could markedly reduce brain edema and improve neurological functions after TBI,showing a neuroprotective effect,possibly through inhibiting TBI-induced autophagy responses.
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