黄芩苷对新生大鼠缺氧缺血性脑损伤神经保护作用的研究

发布时间：2018-06-26 06:13

本文选题：黄芩苷 + 缺氧缺血性脑损伤　；参考：《南昌大学》2012年硕士论文

【摘要】：目的： 1．研究黄芩苷(baicalin, BC)对缺氧缺血性脑损伤(hypoxic-ischemic braindamage, HIBD)新生大鼠脑细胞Fas、FasL表达的影响。 2．比较黄芩苷与神经生长因子(nerve growth factor, NGF)对新生大鼠缺氧缺血性脑损伤的神经保护作用。方法： 1．实验分组：128只7日龄SD大鼠随机分为四组(每组32只)：假手术对照组(Sham组)；缺氧缺血性脑损伤对照组(HIBD组)；神经生长因子对照组(NGF组)；黄芩苷治疗组(BC组)。 2．模型制作及药物干预：参照Rice法制成HIBD模型，其中Sham组仅游离出左颈总动脉，不结扎，不缺氧。NGF组于缺氧缺血(hypoxia-ischemia, HI)、复氧恢复处理后，立刻给予腹腔注射NGF注射液50μg/kg，每天1次，连用3天；BC组于HI、复氧恢复处理后，立刻给予腹腔注射BC注射液16mg/kg，每天1次，连用3天；Sham组和HIBD组每日腹腔注射生理盐水10ml/kg，连用3天。 3．观测指标：观察新生大鼠行为改变、存活率和体重增长；各组均于HI后24小时、48小时、72小时、7天四个时间点(每个时间点各8只)分批取脑组织，HE染色后光镜下观察受损脑组织病理形态改变；S-P免疫组织化学方法测定脑细胞Fas、FasL表达。结果： 1．行为改变：实验大鼠在HI后约30分钟开始出现倦怠，嗜睡，肌张力异常等不同程度的脑损伤症状，复氧后可见大鼠向结扎侧旋转。 2．存活率：Sham组实验大鼠无死亡，，存活率为100%(32/32)；HIBD组死亡5只，存活率为84.38%(27/32)；NGF组死亡2只，存活率为93.75%(30/32)，BC组死亡3只，存活率为90.63%(29/32)。各组间存活率比较，差异无统计学意义(P0.05)。 3．7天体重增长：HIBD组大鼠体重增长最慢，7天体重增长值明显小于其他三组(P0.05)，BC组与NGF组体重增长相近，两组之间比较，差异无统计学意义(P0.05)，但显著落后于Sham组(P0.01)。 4．脑组织病理形态学改变：Sham组大鼠脑细胞结构正常，排列整齐；HIBD组可见脑细胞排列紊乱，细胞水肿明显，部分细胞出现坏死，呈空泡样改变(HI后48小时最为显著)；相同时间点：BC组和NGF组较HIBD组上述改变均明显减轻，而且两者改变相近。 5．脑细胞Fas、FasL表达：Sham组脑细胞可见少量Fas、FasL表达，各时间点差异无统计学意义(P0.05)；其余各组HI后24小时出现Fas、FasL表达，以HI后48小时最明显，之后逐渐减少，持续至7天仍显著高于Sham组，各时间点之间比较，差异有统计学意义(P0.01)；相同时间点：NGF组和BC组之间Fas、FasL表达比较，差异无统计学意义(P0.05)。 6．Fas与FasL相关性分析：新生大鼠HIBD脑细胞Fas表达和FasL表达之间存在直线相关关系(r=0.863，P=0.0000.01)。结论： 1．新生大鼠HIBD脑细胞Fas、FasL表达明显增加，以HI后48小时最为明显，之后逐渐减少，持续至HI后7天仍可见阳性表达，并且正常新生大鼠脑细胞也可见少量Fas、FasL表达。 2．黄芩苷可以通过减少新生大鼠HIBD脑细胞Fas、FasL表达，抑制细胞凋亡，从而发挥脑神经保护作用。 3．黄芩苷对新生大鼠HIBD的神经保护作用与神经生长因子相近，有可能替代神经生长因子成为临床上治疗新生儿HIBD的一种新药物。
[Abstract]:Purpose :

1 . To study the effects of baicalin ( BC ) on the expression of Fas and FasL in brain cells of neonatal rats with hypoxic - ischemic brain damage ( HIBD ) .

2 . To compare the protective effects of baicalin and nerve growth factor ( NGF ) on hypoxic - ischemic brain injury in neonatal rats .

Method :

1 . Experimental group : 128 SD rats were randomly divided into four groups ( 32 in each group ) : sham operation control group ( Sham group ) ;
hypoxic - ischemic brain injury control group ( HIBD group ) ;
Nerve growth factor control group ( NGF group ) ;
baicalin treatment group ( BC group ) .

2 . Model production and drug intervention : The HIBD model was established by reference to Rice method , in which Sham group only free left common carotid artery , no ligation , no hypoxia . NGF group was injected intraperitoneally with 50 渭g / kg NGF injection once a day for 3 days .
BC group was injected with the BC injection at 16 mg / kg once a day for 3 days .
Sham group and HIBD group were injected with normal saline ( 10ml / kg ) daily for 3 days .

3 . Observation index : observe the behavior change , survival rate and body weight gain of neonatal rats ;
The pathological changes of the injured brain tissues were observed under light microscope at 24 hours , 48 hours , 72 hours and 7 days after HI ( 8 rats at each time point ) .
The expression of Fas and FasL in brain cells was measured by S - P immunohistochemical method .

Results :

1 . Behavior change : After HI , the rats began to suffer from various degrees of brain injury , such as lassitude , drowsiness , abnormal muscle tension and so on , and the rats were seen to rotate to the ligation side after hyperbaric oxygenation .

2 . Survival rate : The survival rate was 100 % ( 32 / 32 ) in Sham group .
The survival rate of HIBD group was 84.38 % ( 27 / 32 ) .
The survival rate was 93.75 % ( 30 / 32 ) and the survival rate was 90.63 % ( 29 / 32 ) .

3 . 7 - day body weight gain : the weight gain of the HIBD group was the slowest , and the body weight gain value was significantly lower than that in other three groups ( P0.05 ) . The weight gain of the BC group was similar to that of the NGF group , but the difference was not statistically significant ( P0.05 ) , but the difference was significantly lower than Sham group ( P0.01 ) .

4 . Pathological changes of brain tissue : The brain cells of Sham group were normal and arranged orderly ;
In HIBD group , the brain cells were disordered , cell edema was obvious , some cells appeared necrosis and vacuolar change ( 48 hours after HI was the most significant ) ;
At the same time : BC group and NGF group were significantly lighter than HIBD group , and the changes were similar .

5 . The expression of Fas and FasL in brain cells showed that the expression of Fas and FasL in brain cells of Sham group was not significant ( P0.05 ) .
The expression of Fas and FasL appeared 24 hours after HI , and the expression of Fas and FasL was most obvious at 48 hours after HI . After HI , the expression of Fas and FasL was significantly higher than Sham group after HI , and the difference was significant ( P0.01 ) .
At the same time point : the expression of Fas and FasL between NGF group and BC group was not significant ( P0.05 ) .

6 . Fas and FasL correlation analysis : There was a linear correlation between Fas expression and FasL expression in neonatal rat HIBD brain cells ( r = 0.863 , P = 0.0000 . 01 ) .

Conclusion :

1 . The expression of Fas and FasL in HIBD brain cells was significantly increased in neonatal rats . After HI , the expression of Fas and FasL was gradually decreased and the expression of Fas and FasL was observed in normal neonatal rat brain cells .

2 . baicalin can inhibit the expression of Fas and FasL in HIBD brain cells of neonatal rats , inhibit the apoptosis of cells , and play the role of neuroprotection .

3 . The neuroprotection effect of baicalin on HIBD in neonatal rats is similar to that of nerve growth factor , and it is possible to replace nerve growth factor as a new drug for clinical treatment of HIBD in neonatal rats .
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R722.1

【参考文献】