本文選題：重組人促紅細胞生成素 + 缺氧缺血��； 參考：《安徽醫(yī)科大學》2017年碩士論文

【摘要】：目的:探討重組人促紅細胞生成素對于新生大鼠缺氧缺血性腦損傷受損側(cè)海馬CA1區(qū)NMDA受體NRI亞單位的表達及青春期焦慮行為、學習記憶能力的影響。方法:把7日齡120只Wistar大鼠按隨機數(shù)字表隨機分成對照組、HIBD組、HIBD+rh EPO組,每組均為40只。采用改良的Rice方法制備新生大鼠的HIBD動物模型,術后讓大鼠休息1小時后,把新生大鼠置放在含92%氮氣和8%氧氣的缺氧艙中,讓大鼠持續(xù)缺氧2h。對照組的處理是:僅游離大鼠的右側(cè)頸總動脈,但不結(jié)扎頸總動脈,縫合皮膚后不予缺氧。HIBD+rh EPO組的處理是:在大鼠缺氧缺血后,立即腹腔內(nèi)注射rh EPO,藥物劑量為5000U/Kg。HIBD組與對照組在缺氧缺血后立即腹腔內(nèi)注射與rh EPO體積相等的生理鹽水。在所有實驗大鼠中,按隨機數(shù)字表給每組隨機選擇24只大鼠分別在術后3、12、24、48h四個時間點取大鼠的腦組織標本,并且采用免疫組織化學的染色方法檢測三組大鼠在四個不同時點海馬CA1區(qū)NMDA受體NRI亞單位的表達。每組12只大鼠再分別于35d、49d、63d三個時點完成青春期焦慮行為的實驗研究。每組4只大鼠于35d完成青春期學習記憶能力的實驗研究。結(jié)果:1體重生長發(fā)育情況術前三組實驗大鼠的體重比較結(jié)果無差異統(tǒng)計學意義(P0.05)。從術后到實驗結(jié)束,各組實驗大鼠每日測量體重。術后24h HIBD組的大鼠,體重比手術前有所下降,HIBD組體重明顯低于對照組和治療組,HIBD組與對照組和治療組比較結(jié)果有差異統(tǒng)計學意義(P0.05)。術后48h HIBD組實驗大鼠的體重開始出現(xiàn)緩慢的增長,HIBD組各個時間點的體重均比對照組和治療組的大鼠偏低。直至實驗大鼠生后63d,HIBD組實驗大鼠的體重仍然明顯比照組和治療組低,比較結(jié)果有差異統(tǒng)計學意義(P0.05)。在各個時間段點,對照組實驗大鼠體重和治療組實驗大鼠體重比較,均無差異統(tǒng)計學意義(P0.05)。2免疫組織化學染色HIBD組右側(cè)海馬CA1區(qū)NR1陽性細胞數(shù)比較于HI后3h后開始出現(xiàn)升高,與對照組和治療組比較,比較結(jié)果的差異有統(tǒng)計學意義(P0.05),HIBD組右側(cè)海馬CA1區(qū)NR1陽性細胞數(shù)比較于HI后12h明顯升高,于HI后24h達高峰,3h、12h、24h、48h四個時點右側(cè)受損海馬CA1區(qū)NR1陽性細胞數(shù)與對照組、治療組比較,都高于對照組,比較結(jié)果的差異有統(tǒng)計學意義(P0.05)。治療組3h、12h、24h,48h各時點右側(cè)海馬CA1區(qū)NR1陽性細胞數(shù)低于HIBD組,但高于對照組,三組間結(jié)果的比較,都有差異統(tǒng)計學意義(P0.05)。HIBD組右側(cè)海馬CA1區(qū)NR1陽性細胞數(shù)比較于HI后48h明顯降低,與對照組和治療組比較,比較結(jié)果的差異有統(tǒng)計學意義(P0.05)。3焦慮行為(1)高架十字迷宮實驗P35、P49、P63時三組間入臂總次數(shù)比較,差異均無統(tǒng)計學意義(P0.05)。HIBD組進入開放臂次數(shù)的百分比明顯低于對照組和治療組,差異有統(tǒng)計學意義(P0.05),對照組與治療組進入開放臂次數(shù)的百分比的比較結(jié)果的差異無統(tǒng)計學意義(P0.05),HIBD組進入開放臂滯留時間的百分比明顯低于對照組和治療組,差異有統(tǒng)計學意義(P0.05),對照組與治療組進入開放臂滯留時間的百分比無明顯差異,差異無統(tǒng)計學意義(P0.05)。(2)開場實驗P35、P49、P63時三組實驗大鼠5分鐘內(nèi)水平運動總距離的相互比較結(jié)果的差異不明顯,差異均無統(tǒng)計學意義(P0.05)。HIBD組進入中央?yún)^(qū)總次數(shù)明顯少于對照組和治療組,差異有統(tǒng)計學意義(P0.05),對照組與治療組實驗大鼠進入曠場的中央?yún)^(qū)總次數(shù)的比較結(jié)果無明顯差異,差異無統(tǒng)計學意義(P0.05);HIBD組實驗大鼠在曠場中央?yún)^(qū)停留時間明顯比對照組和治療組低,有明顯的差異,比較結(jié)果的差異有統(tǒng)計學意義(P0.05),對照組與治療組實驗大鼠在曠場中央?yún)^(qū)停留時間的比較明顯差異,比較結(jié)果的差異無統(tǒng)計學意義(P0.05)。4學習記憶能力P35時點時,HIBD組平均逃避潛伏期時間明顯長于對照組和治療組,差異均有統(tǒng)計學意義(P0.05);治療組實驗大鼠平均逃避潛伏期較對照組長,比較結(jié)果的差異有統(tǒng)計學意義(P0.05);P35實驗大鼠在訓練第6天進行穿環(huán)指數(shù)檢測,HIBD組實驗大鼠的穿環(huán)指數(shù)明顯較對照組和治療組低,比較結(jié)果的差異都有統(tǒng)計學意義(P0.05);治療組實驗大鼠的穿環(huán)指數(shù)較對照組低,比較結(jié)果的差異有統(tǒng)計學意義(P0.05)。結(jié)論:1.應用rh EPO干預治療可以減少新生大鼠HIBD海馬CA1區(qū)NMDA受體NRI亞單位的表達。2.HIBD后Wistar大鼠在青春期焦慮情緒增加,合理應用外源性rh EPO進行干預能夠使它的焦慮情緒恢復到正常水平。3.HIBD后Wistar大鼠青春期學習記憶能力下降,合理應用外源性rh EPO進行干預可減輕腦損傷,改善學習記憶能力。但不能恢復至正常水平。
[Abstract]:Objective: To investigate the effect of recombinant human erythropoietin on the expression of NMDA receptor NRI subunit in the hippocampal CA1 region of hypoxic ischemic brain injury in neonatal rats and the influence of learning and memory ability. Methods: 120 Wistar rats of 7 days old were randomly divided into control group, HIBD group and HIBD+rh EPO group, each group was divided into each group. A modified Rice method was used to prepare the HIBD animal model of neonatal rats. After 1 hours of rest, the newborn rats were placed in a hypoxic capsule containing 92% nitrogen and 8% oxygen. The treatment of the rats with sustained anoxic 2h. control group was only the right cervical total arteria of the free rat, but the common carotid artery was not ligated and the skin was sutured without the defect. The treatment of oxygen.HIBD+rh EPO group was: after hypoxic and ischemia of rats, Rh EPO was injected intraperitoneally immediately. The dosage of the drug was 5000U/Kg.HIBD and the control group was injected intraperitoneally with RH EPO immediately after the hypoxic ischemia. In all experimental rats, the random number table was given to each group of 24 rats randomly after the postoperative 3,12,2. 4,48h at four time points was taken from the brain tissue of rats, and immunohistochemical staining was used to detect the expression of NMDA receptor NRI subunit in the hippocampus CA1 region of four different groups of rats. 12 rats in each group completed the experimental study of adolescent anxiety behavior at the three time points of 35d, 49D and 63d. 4 rats in each group were completed in 35d. Experimental study on the learning and memory ability of puberty results: 1 the weight comparison of the three groups before the operation was not statistically significant (P0.05). From postoperative to the end of the experiment, the weight of the rats in each group was measured every day. The weight of the rats in group 24h HIBD after operation was lower than that before the operation, and the body weight of the group HIBD was significantly lower than that of the group. In the control group and the treatment group, there was a significant difference between the HIBD group and the control group and the treatment group (P0.05). The weight of the experimental rats in group 48h HIBD began to grow slowly, and the weight of each time point in group HIBD was lower than that of the control group and the treatment group. The weight of the rats in the group of HIBD was still 63d, and the weight of the experimental rats in the group HIBD was still the same. There was a significant difference in the comparison between the group and the treatment group (P0.05). At each time point, the weight of the experimental rats in the control group and the weight of the experimental group were not statistically significant (P0.05) the number of NR1 positive cells in the right hippocampal CA1 region of the HIBD group of.2 immuno histochemistry was higher than that of the 3h after HI. Compared with the control group and the treatment group, the difference of the results was statistically significant (P0.05). The number of NR1 positive cells in the right hippocampal CA1 area in the right hippocampus of the HIBD group was significantly higher than that after HI, and the 24h reached the peak at the HI after HI, 3h, 12h, 24h, and the number of positive cells in the right damaged hippocampus at the four point of 48h was compared with the control group, and the comparison was higher than the control group. The comparison was compared with the control group, and the comparison was higher than the control group. The difference of the results was statistically significant (P0.05). The number of NR1 positive cells in the right hippocampal CA1 region of the treatment group was lower than that of the HIBD group at the time points of 3H, 12h, 24h and 48h, but it was higher than the control group. The comparison of the results between the three groups was statistically significant (P0.05) in the CA1 region of the hippocampus, the number of NR1 Yang cells in the CA1 region of the hippocampus was significantly lower than that in the control and treatment groups. Comparison, the difference of the results was statistically significant (P0.05).3 anxiety behavior (1) the elevated cross maze test of P35, P49, P63, the total number of the three groups, the difference was not statistically significant (P0.05) the percentage of the.HIBD group entered the number of open arms was significantly lower than the control group and the treatment group, the difference was statistically significant (P0.05), the control group and the treatment. There was no significant difference in the percentage of the percentage of the group entering the open arm (P0.05). The percentage of the entry time in the open arm of the group HIBD was significantly lower than that in the control group and the treatment group (P0.05). There was no significant difference between the control group and the treatment group in the opening time of the open arm. There was no significant difference in the difference between the control group and the treatment group. P0.05) (2) (2) there was no significant difference between the three groups of experimental rats within 5 minutes of the opening experiment at P35, P49, and P63. The difference was not statistically significant (P0.05) in group.HIBD, the total frequency of entering the central area was significantly less than that of the control group and the treatment group, and the difference had the significance of the overall planning (P0.05), the control group and the treatment group entered the open area. There was no significant difference between the total number of times in the central area of the central area (P0.05). The retention time of the experimental rats in the HIBD group was significantly lower than the control group and the treatment group. There was a significant difference between the experimental group and the treatment group. The difference of the results was statistically significant (P0.05). The control group and the treatment group were in the central area of the open field central area. There was no significant difference. There was no significant difference in the comparison results (P0.05).4 learning and memory ability P35 time point, the average escape latency time of group HIBD was significantly longer than that of the control group and the treatment group, the difference was statistically significant (P0.05); the average escape latency of the experimental group was longer than that of the control group, and the difference of the results was statistically significant. Meaning (P0.05); P35 experimental rats were tested for sixth days of piercing index. The ring index of group HIBD rats was significantly lower than that of the control group and the treatment group, and the difference of the results was statistically significant (P0.05); the ring index of the experimental rats was lower than that of the control group, and the difference between the results was statistically significant (P0.05). Conclusion: the 1. application The intervention therapy of RH EPO can reduce the expression of NMDA receptor NRI subunit in the HIBD hippocampus CA1 region of neonatal rats and increase the anxiety mood of Wistar rats in puberty. The rational application of exogenous RH EPO to intervene can make its anxiety restore to the normal level.3.HIBD Wistar rats' learning and memory ability decline, reasonable application. Exogenous RH EPO intervention can reduce brain damage and improve learning and memory ability, but it can not return to normal level.

【學位授予單位】：安徽醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R742

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