β-七叶皂苷钠对SSST大鼠脑水肿、血脑屏障及AQP-4的影响

发布时间：2018-04-30 16:06

本文选题：SSST + AQP-4　；参考：《南华大学》2014年硕士论文

【摘要】：目的：研究大鼠上矢状窦中1/3及其回流静脉结扎（SSS+RV）后脑水肿、AQP-4及血脑屏障破坏程度三者之间的关系；并探讨β-七叶皂苷钠是否能通过调节SSS+RV后窦旁脑组织中AQP-4的表达而减轻脑水肿程度。方法：96只健康雄性SD大鼠，按照完全随机原则分为假手术组(ShamOperation，SO组)6只；上矢状窦中l/3及其回流静脉结扎组(SSS+RV组/SR组),SR组按处死时间点即结扎后6h、12h、24h、48h、72h分为5亚组，每个亚组6只；干预组60只分为生理盐水干预组(NS+SSS+RV组/NS组)和β-七叶皂苷钠干预组（SA+SSS+RV组/SA组)，各组又细分为6h、12h、24h、48h、72h组5个亚组，每亚组6只。各组大鼠于相应时间点处死，处死前观察大鼠神经功能缺损程度，光镜下观察窦旁脑组织病理变化、干湿重法检测窦旁脑组织含水量、检测窦旁脑组织EB漏出量反映血-脑屏障通透性、免疫组化法检测AQP-4蛋白的表达。结果：1）SO组未出现神经功能缺损，而SR、NS组均于结扎SSS中1/3及其回流静脉后6h开始出现神经功能障碍，且随着时间的延长，神经功能缺损越严重，于24h神经功能评分最高，此后，神经功能评分逐渐降低，72h神经功能评分较24h明显降低；SA各亚组较SR、NS组神经功能缺损程度明显减轻，且差异具有统计学意义（P0.05）； 2）SR组、NS组大鼠SSS+RV后6h，HE染色开始出现神经元水肿、变性、坏死，Nissl染色显示尼氏小体数量减少，二者均于24h时最明显，此后随时间延长神经元变性较前好转、尼氏体数量逐渐增加，与SR、NS组比较，SA组上述变化明显减轻； 3）大鼠SSS+RV6h时，各组脑组织含水量开始增加（与SO组比较，P0.05），并于结扎后24h达高峰，此后随时间延长逐渐降低，72h时脑组织含水量较24h明显减少；而予以生理盐水干预组（NS组），各时间点相应亚组脑组织含水量与SR组中对应亚组比较，无明显差异，给予β-七叶皂苷钠干预组（SA组）中，脑组织含水量则明显减少（P0.05）； 4）大鼠SSS+RV6h时，各组AQP-4表达量开始增加（与SO组比较，P0.05），于结扎后24h达高峰，随后其表达逐渐减少，72h时AQP-4含量较24h明显减少；而予以生理盐水干预组（NS组），各时间点相应亚组脑组织AQP-4蛋白含量与SR组中对应亚组比较，无明显差异，给予β-七叶皂苷钠干预组（SA组）中，AQP-4表达量则明显减低（P0.05）； 5）大鼠SSS+RV6h时，各组窦旁脑组织EB漏出量开始增加（与SO组比较，P0.05），并于结扎后24h窦旁脑组织EB含量最高，此后随时间延长逐渐减少,72h时EB含量较24h明显减少；而予以生理盐水干预组（NS组），各时间点相应亚组脑组织EB含量与SR组中对应亚组比较，无明显差异，给予β-七叶皂苷钠干预组（SA组）中，脑组织EB漏出量则明显减少（P0.05）； 6）大鼠SSS+RV结扎后，，脑含水量与EB含量呈显著正相关(r=0.796,p0.01)，脑含水量与AQP-4蛋白的表达呈显著正相关(r=0.773,p0.01)，且EB含量与AQP-4蛋白的表达也呈显著正相关(r=0.928,p0.01)。结论：1）大鼠SSS中1/3及其回流静脉结扎后，脑水肿程度、AQP-4含量及血脑屏障破坏三者之间密切相关； 2）β-七叶皂苷钠可通过下调脑组织中AQP-4的表达而减轻血脑屏障的破坏程度、减轻脑水肿程度。
[Abstract]:Objective: To investigate the relationship between the cerebral edema after 1/3 and refluxing venous ligation (SSS+RV) in the superior sagittal sinus of rats, the relationship between AQP-4 and the damage degree of blood brain barrier (BBB) three, and to explore whether beta seven leaf saponins can reduce the degree of cerebral edema by regulating the expression of AQP-4 in the Paranal brain tissue of SSS+RV.
Methods: 96 healthy male SD rats were divided into 6 rats in the sham operation group (ShamOperation, group SO) according to the principle of complete randomization, l/3 in the upper sagittal sinus and the reflux vein ligature group (group /SR group SSS+RV). The SR group was divided into 5 subgroups, 12h, 24h, 48H, and 6 in each subgroup, and 60 in the intervention group. Group SSS+RV /NS group) and beta seven leaf saponin sodium intervention group (group /SA group SA+SSS+RV), each group was subdivided into 6h, 12h, 24h, 48h, and 72h group 5 subgroups, 6 in each subgroup. The rats were killed at the corresponding time point, the degree of nerve function defect was observed in the rats before death, the pathological changes of paranantral brain tissue were observed under light microscope, and the water content of paranantral brain tissue was detected by dry wet weight method. The leakage of EB in the paranasal brain tissue was detected to reflect the permeability of blood brain barrier, and the expression of AQP-4 protein was detected by immunohistochemistry.
Results: 1) there was no nerve function defect in SO group, and SR, NS group began to appear nerve dysfunction after 1/3 and its reflux vein in SSS, and with the prolongation of time, the nervous function defect was more serious, the neurological function score of 24h was the highest. After that, the neurological function score was gradually reduced, 72h nerve function score was lower than 24h, SA, and SA respectively. Compared with group SR and group NS, the degree of neurological deficit was significantly reduced, and the difference was statistically significant (P0.05).
2) SR group, group NS rats after SSS+RV 6h, HE staining began to appear neuron edema, degeneration, necrosis, Nissl staining showed the number of Nissl corpuscles decreased, the most obvious in the 24h, after the time prolonged neuronal degeneration, the number of Nissl body gradually increased, compared with SR, NS group, SA group changes obviously reduced.
3) when the rats were SSS+RV6h, the water content of the brain tissues began to increase (compared with the SO group, P0.05), and after the ligation, the 24h reached the peak, and then gradually decreased with the time, and the water content of the brain tissue decreased obviously at 72h, while the physiological saline intervention group (NS group), the water content in the corresponding subgroups of the corresponding subgroups of each time point was compared with the corresponding subgroup in the SR group. Obviously, the water content in brain tissue of the intervention group (group SA) was significantly decreased (P0.05) after giving the beta seven saponin sodium intervention group.
4) when rats were SSS+RV6h, the expression of AQP-4 began to increase (compared with that of group SO, P0.05). After ligation, 24h reached its peak, and then its expression decreased gradually, and the content of AQP-4 decreased obviously at 72h. The content of AQP-4 protein in the corresponding subgroups of the corresponding subgroups of the corresponding sub group of the physiological saline intervention group (NS group) was compared with the corresponding subgroup in the SR group, and there was no significant difference. In the intervention group (group SA), the expression of AQP-4 decreased significantly (P0.05) in the intervention group (seven groups).
5) the EB leakage of paranantral tissues in the parantral tissues of the rats began to increase (compared with that of the SO group, P0.05), and the content of EB in the paranus paranus was the highest after ligation, and then gradually decreased with time, and the content of EB in 72h decreased significantly than that in 72h, while the physiological saline intervention group (NS group), and the EB content of the corresponding subgroup of brain tissue in the corresponding subgroup of each time point and the SR group, was given. There was no significant difference between the subgroup and the group treated with beta - seven saponin sodium (group SA). The leakage of EB in brain tissue was significantly reduced (P0.05).
6) after the ligation of SSS+RV, the water content of the brain was significantly positively correlated with the content of EB (r=0.796, P0.01). The water content of the brain was positively correlated with the expression of AQP-4 protein (r=0.773, P0.01), and the content of EB was also positively correlated with the expression of AQP-4 protein (r=0.928, P0.01).
Conclusion: 1) there was a close correlation between the degree of brain edema, AQP-4 content and blood-brain barrier destruction in SSS rats after 1/3 and their veins were ligated.
2) beta seven sodium saponin can reduce the damage of blood-brain barrier and reduce the degree of brain edema by downregulating the expression of AQP-4 in brain tissue.

【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742

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