临床儿科杂志
臨床兒科雜誌
림상인과잡지
2015年
4期
368-372
,共5页
徐发林%王彩红%张彦华%郭佳佳%程慧清
徐髮林%王綵紅%張彥華%郭佳佳%程慧清
서발림%왕채홍%장언화%곽가가%정혜청
枸橼酸咖啡因%5-溴脱氧尿嘧啶核苷%活化半胱氨酸天冬氨酸蛋白酶-3%大鼠
枸櫞痠咖啡因%5-溴脫氧尿嘧啶覈苷%活化半胱氨痠天鼕氨痠蛋白酶-3%大鼠
구연산가배인%5-추탈양뇨밀정핵감%활화반광안산천동안산단백매-3%대서
caffeine citrate%5-bromo-2’-deoxyuridine%cleaved caspase-3%rat
目的：研究枸橼酸咖啡因(CC)对新生大鼠缺氧缺血性脑损伤(HIBD)后神经细胞增生与凋亡及长期学习记忆能力的影响。方法7日龄SD新生大鼠随机分为假手术组、HIBD组、CC组，每组16只；HIBD组及CC组经左颈总动脉结扎并缺氧制作HIBD模型，CC组在HI前、HI后0 min、24 h、48 h、72 h给予CC 20 mg/kg腹腔注射，假手术组和HIBD组分别在同一时间点给予等量生理盐水腹腔注射；同时从生后10日龄起腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)标记新生细胞，剂量50 mg/kg，每12 h 1次，共5次。12日龄时每组随机选取8只大鼠处死，免疫组织化学染色检测海马齿状回颗粒下层5-溴脱氧尿嘧啶核苷(BrdU)和海马CA1区活化半胱氨酸天冬氨酸蛋白酶(活化Caspase-3)的表达情况， TUNEL法测定海马CA1区神经细胞凋亡情况，其余各组大鼠28日龄时行Y迷宫学习和记忆能力测试。结果三组新生大鼠脑组织中均可见BrdU阳性细胞，差异有统计学意义(F=101.38，P<0.01)；HIBD组、CC组BrdU阳性细胞数均较假手术组增多，差异有统计学意义(P<0.05)。三组新生大鼠海马CA1区均可见活化 Caspase-3阳性细胞，差异有统计学意义(F=379.77，P<0.01)；CC组活化 Caspase-3阳性细胞较HIBD组显著减少，但仍多于假手术组，差异有统计学意义(P<0.05)。三组新生大鼠海马CA1区中均可见TUNEL阳性细胞，差异有统计学意义(F=505.92，P<0.01)，以HIBD组最多，假手术组最少，两两比较差异均有统计学意义(P<0.05)。Y迷宫实验结果，各组大鼠达标所需训练总次数的差异有统计学意义(F=32.05，P<0.01)，以HIBD组所需训练次数最多。24 h后正确反应率在三组间的差异也有统计学意义(F=24.99，P<0.01)，以HIBD组大鼠正确反应率最低。结论枸橼酸咖啡因可以改善HIBD大鼠长期学习记忆力，其机制可能与通过减少缺氧缺血后神经细胞的凋亡有关。
目的：研究枸櫞痠咖啡因(CC)對新生大鼠缺氧缺血性腦損傷(HIBD)後神經細胞增生與凋亡及長期學習記憶能力的影響。方法7日齡SD新生大鼠隨機分為假手術組、HIBD組、CC組，每組16隻；HIBD組及CC組經左頸總動脈結扎併缺氧製作HIBD模型，CC組在HI前、HI後0 min、24 h、48 h、72 h給予CC 20 mg/kg腹腔註射，假手術組和HIBD組分彆在同一時間點給予等量生理鹽水腹腔註射；同時從生後10日齡起腹腔註射5-溴脫氧尿嘧啶覈苷(BrdU)標記新生細胞，劑量50 mg/kg，每12 h 1次，共5次。12日齡時每組隨機選取8隻大鼠處死，免疫組織化學染色檢測海馬齒狀迴顆粒下層5-溴脫氧尿嘧啶覈苷(BrdU)和海馬CA1區活化半胱氨痠天鼕氨痠蛋白酶(活化Caspase-3)的錶達情況， TUNEL法測定海馬CA1區神經細胞凋亡情況，其餘各組大鼠28日齡時行Y迷宮學習和記憶能力測試。結果三組新生大鼠腦組織中均可見BrdU暘性細胞，差異有統計學意義(F=101.38，P<0.01)；HIBD組、CC組BrdU暘性細胞數均較假手術組增多，差異有統計學意義(P<0.05)。三組新生大鼠海馬CA1區均可見活化 Caspase-3暘性細胞，差異有統計學意義(F=379.77，P<0.01)；CC組活化 Caspase-3暘性細胞較HIBD組顯著減少，但仍多于假手術組，差異有統計學意義(P<0.05)。三組新生大鼠海馬CA1區中均可見TUNEL暘性細胞，差異有統計學意義(F=505.92，P<0.01)，以HIBD組最多，假手術組最少，兩兩比較差異均有統計學意義(P<0.05)。Y迷宮實驗結果，各組大鼠達標所需訓練總次數的差異有統計學意義(F=32.05，P<0.01)，以HIBD組所需訓練次數最多。24 h後正確反應率在三組間的差異也有統計學意義(F=24.99，P<0.01)，以HIBD組大鼠正確反應率最低。結論枸櫞痠咖啡因可以改善HIBD大鼠長期學習記憶力，其機製可能與通過減少缺氧缺血後神經細胞的凋亡有關。
목적：연구구연산가배인(CC)대신생대서결양결혈성뇌손상(HIBD)후신경세포증생여조망급장기학습기억능력적영향。방법7일령SD신생대서수궤분위가수술조、HIBD조、CC조，매조16지；HIBD조급CC조경좌경총동맥결찰병결양제작HIBD모형，CC조재HI전、HI후0 min、24 h、48 h、72 h급여CC 20 mg/kg복강주사，가수술조화HIBD조분별재동일시간점급여등량생리염수복강주사；동시종생후10일령기복강주사5-추탈양뇨밀정핵감(BrdU)표기신생세포，제량50 mg/kg，매12 h 1차，공5차。12일령시매조수궤선취8지대서처사，면역조직화학염색검측해마치상회과립하층5-추탈양뇨밀정핵감(BrdU)화해마CA1구활화반광안산천동안산단백매(활화Caspase-3)적표체정황， TUNEL법측정해마CA1구신경세포조망정황，기여각조대서28일령시행Y미궁학습화기억능력측시。결과삼조신생대서뇌조직중균가견BrdU양성세포，차이유통계학의의(F=101.38，P<0.01)；HIBD조、CC조BrdU양성세포수균교가수술조증다，차이유통계학의의(P<0.05)。삼조신생대서해마CA1구균가견활화 Caspase-3양성세포，차이유통계학의의(F=379.77，P<0.01)；CC조활화 Caspase-3양성세포교HIBD조현저감소，단잉다우가수술조，차이유통계학의의(P<0.05)。삼조신생대서해마CA1구중균가견TUNEL양성세포，차이유통계학의의(F=505.92，P<0.01)，이HIBD조최다，가수술조최소，량량비교차이균유통계학의의(P<0.05)。Y미궁실험결과，각조대서체표소수훈련총차수적차이유통계학의의(F=32.05，P<0.01)，이HIBD조소수훈련차수최다。24 h후정학반응솔재삼조간적차이야유통계학의의(F=24.99，P<0.01)，이HIBD조대서정학반응솔최저。결론구연산가배인가이개선HIBD대서장기학습기억력，기궤제가능여통과감소결양결혈후신경세포적조망유관。
ObjectiveTo investigate the effects of caffeine citrate (CC) on the neuronal proliferation and apoptosis and long-term learning ability in neonatal rats with hypoxia-ischemic brain damage (HIBD).MethodsForty-eight 7-day-old Sprague-Dawley neonatal rats were randomly divided into 3 groups: sham operation group (n=16), HIBD group (n=16), HIBD + caffeine citrate group (CC group,n=16). Rats in HIBD and CC groups received ligation of left common carotid artery followed by 2 hours of hypoxia to establish HIBD model. Rats in CC group were injected intraperitoneally with CC (20 mg/kg) before and at 0 min, 24 h, 48 h, and 72 h after hypoxia-ischemic (HI), and rats in the other two groups were injected intraperitoneally with an equal volume of normal saline at the corresponding time. Meanwhile, from postnatal day 10, each rat was injected intraperitoneally with 5-bromo-2’-de-oxyuridine (BrdU) (50 mg/kg) for 5 consecutive times, once every 12 h. On postnatal day 12, BrdU in the hippocampal dentate gyrus and cleaved caspase-3 in the hippocampal CA1 area were detected by immunohistochemistry, and neuronal apoptosis in hippocampal CA1 area were detected by TUNEL staining. On postnatal day 28, long-term learning and memory ability of rats was tested by Y maze.ResultsThere was signiifcant difference in the number of BrdU-positive cells in brain tissues of rats among three groups (F=101.38,P<0.01). The BrdU-positive cells in HIBD group and CC group were signiifcantly more than those in sham operation group (P<0.05). There was signiifcant difference in the number of cleaved caspase-3-positive cells in hippocampal CA1 area among three groups (F=379.77,P<0.01). The cleaved caspase-3-positive cells in CC group were signiif-cantly fewer than those in HIBD group but signiifcantly more than those in sham operation group (P<0.05). The TUNEL-pos-itive cells in hippocampal CA1 area were signiifcantly different among three groups (F=505.92,P<0.01) which was most in HIBD group and fewest in sham operation group and signiifcant difference was found through multiple comparison (P<0.05). The total learning number of avoiding electric shock tested by Y maze was signiifcantly different among three groups (F=32.05, P<0.01) which was most in HIBD group. Correct response rate was significantly different among three groups (F=24.99, P<0.01) which was lowest in HIBD group.ConclusionsCaffeine citrate can improve the ability of long-term learning and memory in neonatal rats with hypoxia-ischemic brain damage, the mechanism of which may be related to reducing the neuronal apoptosis after hypoxia ischemia.
