rhEPO对缺氧缺血性脑损伤新生大鼠海马CA1区NMDA受体NRI亚单位的表达及青春期焦虑行为和学习记忆的影响

发布时间：2018-04-22 20:29

本文选题：重组人促红细胞生成素 + 缺氧缺血　；参考：《安徽医科大学》2017年硕士论文

【摘要】：目的:探讨重组人促红细胞生成素对于新生大鼠缺氧缺血性脑损伤受损侧海马CA1区NMDA受体NRI亚单位的表达及青春期焦虑行为、学习记忆能力的影响。方法:把7日龄120只Wistar大鼠按随机数字表随机分成对照组、HIBD组、HIBD+rh EPO组,每组均为40只。采用改良的Rice方法制备新生大鼠的HIBD动物模型,术后让大鼠休息1小时后,把新生大鼠置放在含92%氮气和8%氧气的缺氧舱中,让大鼠持续缺氧2h。对照组的处理是:仅游离大鼠的右侧颈总动脉,但不结扎颈总动脉,缝合皮肤后不予缺氧。HIBD+rh EPO组的处理是:在大鼠缺氧缺血后,立即腹腔内注射rh EPO,药物剂量为5000U/Kg。HIBD组与对照组在缺氧缺血后立即腹腔内注射与rh EPO体积相等的生理盐水。在所有实验大鼠中,按随机数字表给每组随机选择24只大鼠分别在术后3、12、24、48h四个时间点取大鼠的脑组织标本,并且采用免疫组织化学的染色方法检测三组大鼠在四个不同时点海马CA1区NMDA受体NRI亚单位的表达。每组12只大鼠再分别于35d、49d、63d三个时点完成青春期焦虑行为的实验研究。每组4只大鼠于35d完成青春期学习记忆能力的实验研究。结果:1体重生长发育情况术前三组实验大鼠的体重比较结果无差异统计学意义(P0.05)。从术后到实验结束,各组实验大鼠每日测量体重。术后24h HIBD组的大鼠,体重比手术前有所下降,HIBD组体重明显低于对照组和治疗组,HIBD组与对照组和治疗组比较结果有差异统计学意义(P0.05)。术后48h HIBD组实验大鼠的体重开始出现缓慢的增长,HIBD组各个时间点的体重均比对照组和治疗组的大鼠偏低。直至实验大鼠生后63d,HIBD组实验大鼠的体重仍然明显比照组和治疗组低,比较结果有差异统计学意义(P0.05)。在各个时间段点,对照组实验大鼠体重和治疗组实验大鼠体重比较,均无差异统计学意义(P0.05)。2免疫组织化学染色HIBD组右侧海马CA1区NR1阳性细胞数比较于HI后3h后开始出现升高,与对照组和治疗组比较,比较结果的差异有统计学意义(P0.05),HIBD组右侧海马CA1区NR1阳性细胞数比较于HI后12h明显升高,于HI后24h达高峰,3h、12h、24h、48h四个时点右侧受损海马CA1区NR1阳性细胞数与对照组、治疗组比较,都高于对照组,比较结果的差异有统计学意义(P0.05)。治疗组3h、12h、24h,48h各时点右侧海马CA1区NR1阳性细胞数低于HIBD组,但高于对照组,三组间结果的比较,都有差异统计学意义(P0.05)。HIBD组右侧海马CA1区NR1阳性细胞数比较于HI后48h明显降低,与对照组和治疗组比较,比较结果的差异有统计学意义(P0.05)。3焦虑行为(1)高架十字迷宫实验P35、P49、P63时三组间入臂总次数比较,差异均无统计学意义(P0.05)。HIBD组进入开放臂次数的百分比明显低于对照组和治疗组,差异有统计学意义(P0.05),对照组与治疗组进入开放臂次数的百分比的比较结果的差异无统计学意义(P0.05),HIBD组进入开放臂滞留时间的百分比明显低于对照组和治疗组,差异有统计学意义(P0.05),对照组与治疗组进入开放臂滞留时间的百分比无明显差异,差异无统计学意义(P0.05)。(2)开场实验P35、P49、P63时三组实验大鼠5分钟内水平运动总距离的相互比较结果的差异不明显,差异均无统计学意义(P0.05)。HIBD组进入中央区总次数明显少于对照组和治疗组,差异有统计学意义(P0.05),对照组与治疗组实验大鼠进入旷场的中央区总次数的比较结果无明显差异,差异无统计学意义(P0.05);HIBD组实验大鼠在旷场中央区停留时间明显比对照组和治疗组低,有明显的差异,比较结果的差异有统计学意义(P0.05),对照组与治疗组实验大鼠在旷场中央区停留时间的比较明显差异,比较结果的差异无统计学意义(P0.05)。4学习记忆能力P35时点时,HIBD组平均逃避潜伏期时间明显长于对照组和治疗组,差异均有统计学意义(P0.05);治疗组实验大鼠平均逃避潜伏期较对照组长,比较结果的差异有统计学意义(P0.05);P35实验大鼠在训练第6天进行穿环指数检测,HIBD组实验大鼠的穿环指数明显较对照组和治疗组低,比较结果的差异都有统计学意义(P0.05);治疗组实验大鼠的穿环指数较对照组低,比较结果的差异有统计学意义(P0.05)。结论:1.应用rh EPO干预治疗可以减少新生大鼠HIBD海马CA1区NMDA受体NRI亚单位的表达。2.HIBD后Wistar大鼠在青春期焦虑情绪增加,合理应用外源性rh EPO进行干预能够使它的焦虑情绪恢复到正常水平。3.HIBD后Wistar大鼠青春期学习记忆能力下降,合理应用外源性rh EPO进行干预可减轻脑损伤,改善学习记忆能力。但不能恢复至正常水平。
[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.

【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R742

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