本文選題：金釵石斛總生物堿 + 散發(fā)性老年癡呆�。� 參考：《遵義醫(yī)學(xué)院》2017年碩士論文

【摘要】：目的:探索金釵石斛總生物堿(DNLA)對鏈脲佐菌素(STZ)誘導(dǎo)的大鼠學(xué)習(xí)記憶減退模型的作用。方法:將50只雄性SD大鼠隨機(jī)分成5組:空白對照組(Control)、空白給藥組(Control+DNLA40)、模型組(STZ)、低劑量組(STZ+DNLA20)和高劑量組(STZ+DNLA40)。每組10只大鼠。對大鼠進(jìn)行麻醉,后將其固定于立體定位儀上,定位注射時(shí)的坐標(biāo)位置:前囟后0.8 mm,左右旁開1.5 mm,顱骨下3.6mm。使用微量進(jìn)樣器對側(cè)腦室緩慢注STZ溶液(70.5μg/μL),以1μL/min速度注入,5 min注完,留針5 min,然后對另一側(cè)側(cè)腦室進(jìn)行相同操作,最終使每只用于模型的大鼠接受STZ劑量為3 mg/kg,制備散發(fā)性老年癡呆(sporadic Alzheimer’s Disease,SAD)動(dòng)物模型。模型制備完成后,對需要給藥的實(shí)驗(yàn)組每天定時(shí)給予20 mg/kg或40 mg/kg的DNLA,連續(xù)28天;而模型組和空白對照組則給予等體積的溶媒處理。本研究使用Morris水迷宮法檢測單次雙側(cè)側(cè)腦室注射鏈脲佐菌素(ICV-STZ)對大鼠的空間學(xué)習(xí)記憶功能的影響,從ICV-STZ后第23天開始連續(xù)進(jìn)行5天,并在該測試結(jié)束后第二天,進(jìn)行空間探索實(shí)驗(yàn)。經(jīng)蘇木素-伊紅染色及尼氏染色,光鏡觀察DNLA對大鼠海馬神經(jīng)元密度和形態(tài)的影響。采用Western blot法檢測大鼠海馬內(nèi)Tau蛋白,胰島素受體(IR),胰島素底物-1(IRS-1),蛋白激酶B(Akt)和糖原合成激酶-3β(GSK-3β)的蛋白表達(dá)水平和磷酸化程度,并檢測了蛋白磷酸酶2-α(PP2A-α)的蛋白表達(dá)水平。結(jié)果:行為學(xué)實(shí)驗(yàn)中,通過Morris水迷宮實(shí)驗(yàn)測試發(fā)現(xiàn),模型組的大鼠的逃逸潛伏期較空白對照組的從第4天開始表現(xiàn)為的顯著升高;給藥DNLA后,高劑量組的大鼠逃逸潛伏期較模型組在第五天出現(xiàn)顯著降低;而低劑量組的大鼠逃逸潛伏期較模型組無顯著差異;另外,對大鼠的游泳速度測試,各組大鼠未見顯著差異。組織切片實(shí)驗(yàn)中,經(jīng)HE和Nissl染色發(fā)現(xiàn),模型組的大鼠海馬CA3區(qū)的神經(jīng)元數(shù)量顯著減少,細(xì)胞排列層次減少,部分細(xì)胞出現(xiàn)深染、核固縮;給藥DNLA后,高劑量組的大鼠海馬CA3區(qū)正常神經(jīng)元數(shù)量較模型組明顯增多,排列層次增加,且未見明顯的細(xì)胞深染及核固縮現(xiàn)象;低劑量組的大鼠海馬CA3區(qū)亦可觀察到正常神經(jīng)元數(shù)量較模型組增多,深染及核固縮的細(xì)胞減少;免疫印跡實(shí)驗(yàn)中,經(jīng)Western blot法檢測發(fā)現(xiàn),在模型組的大鼠海馬內(nèi),與正常組比較,IR在酪氨酸1361位點(diǎn)和GSK-3β在絲氨酸9位點(diǎn)的磷酸化程度降低,GSK-3β在酪氨酸216位點(diǎn)的磷酸化水平以及PP2A-α蛋白水平無顯著改變;IRS-1在絲氨酸616位點(diǎn),Akt在絲氨酸473位點(diǎn),Tau蛋白在絲氨酸199、396、404、422位點(diǎn)、蘇氨酸616位點(diǎn)的磷酸化水平升高。給藥DNLA后,在高劑量組的大鼠海馬內(nèi),與模型組的大鼠比較,IR在酪氨酸1361位點(diǎn)和GSK-3β在絲氨酸9位點(diǎn)的磷酸化程度與模型組比較增加,GSK-3β在酪氨酸216位點(diǎn)的磷酸化水平以及PP2A-α蛋白水平無顯著改變;IRS-1在絲氨酸616位點(diǎn),Akt在絲氨酸473位點(diǎn),Tau蛋白在絲氨酸199、396、404、422位點(diǎn)、蘇氨酸616位點(diǎn)的磷酸化水平降低;而在低劑量組的大鼠海馬內(nèi),與模型組相比較,可見Tau蛋白在絲氨酸404、蘇氨酸231位點(diǎn)的磷酸化水平較模型組降低,Akt在絲氨酸473位點(diǎn)和GSK-3β在絲氨酸9位點(diǎn)磷酸化水平降低。結(jié)論:DNLA可減輕STZ誘導(dǎo)的大鼠學(xué)習(xí)記憶減退及神經(jīng)元損傷,該作用與DNLA改善STZ導(dǎo)致的大鼠海馬中的Tau蛋白過度磷酸化及胰島素信號通路障礙有關(guān)。
[Abstract]:Objective: To explore the effect of Dendrobium nobile total alkaloid (DNLA) on the learning and memory impairment model induced by streptozotocin (STZ) in rats. Methods: 50 male SD rats were randomly divided into 5 groups: blank control group (Control), blank administration group (Control+DNLA40), model group (STZ), low dose group (STZ+DNLA20) and high dose group (STZ+DNLA40). 10 rats in each group were large. Rats were anesthetized, then fixed to the stereotaxis, positioning the coordinates of the coordinates: 0.8 mm in the anterior Fontane, 1.5 mm beside the side of the skull, and the slow injection of STZ solution (70.5 mu g/ mu L) to the lateral ventricle using a micro injector for the lower 3.6mm. of the skull, 5 Min injection, 5 min, and the same exercise on the other side of the ventricle. As a result, each rat used for the model was eventually accepted STZ 3 mg/kg to prepare sporadic Alzheimer's (sporadic Alzheimer 's Disease, SAD) animal model. After the model preparation was completed, the experimental group needed to be given a dose of 20 mg/kg or 40 mg/kg DNLA for even 28 days, while the model group and the blank control group were given equal volume. The effect of single lateral lateral ventricle injection of streptozotocin (ICV-STZ) on the spatial learning and memory function of rats was detected by Morris water maze. The space exploration was carried out for 5 days from twenty-third days after ICV-STZ, and the space exploration experiment was carried out at the end of the test after the end of the test. The effects of DNLA on the density and morphology of hippocampal neurons in rats were observed. The protein expression level and phosphorylation level of Tau protein, insulin receptor (IR), insulin substrate -1 (IRS-1), protein kinase B (Akt) and glycogen kinase -3 beta (GSK-3 beta) were detected by Western blot method, and the protein phosphatase 2- alpha protein was detected. Results: in the behavior test, the Morris water maze test showed that the escape latency of the rats in the model group was significantly higher than that in the blank control group from fourth days. After DNLA, the escape latency of the rats in the high dose group was significantly lower than that of the model group at fifth days, while the rats in the low dose group fled. There was no significant difference between the model group and the model group. In addition, there was no significant difference in the swimming speed test of rats. In the tissue section experiment, HE and Nissl staining showed that the number of neurons in the hippocampus CA3 area of the rat model group decreased significantly, the level of cell arrangement decreased, some cells appeared deep dyed and nuclear pyknosis; after the administration of DNLA, a high dose of drug was used. The number of normal neurons in the hippocampal CA3 area of rats in the dose group increased significantly, the arrangement level increased, and there was no obvious cell deep staining and nuclear condensation. The number of normal neurons in the hippocampus CA3 area of the low dose group could also be observed to be more than the model group, and the deep staining and nuclear pyknosis cells decreased; in Western blot experiments, Western blo T assay found that in the rat hippocampus of the model group, the phosphorylation of IR at the tyrosine 1361 site and GSK-3 beta at the serine 9 site was lower than that in the normal group. The phosphorylation level of GSK-3 beta at the tyrosine 216 site and the level of PP2A- alpha protein were not significantly changed; IRS-1 was at the 616 site of serine, Akt at the serine 473 site, and Tau protein in silk The phosphorylation level of the threonine 616 site was increased at the 199396404422 site of the ammonia acid site. After DNLA, the phosphorylation of IR at the tyrosine 1361 site and the GSK-3 beta at the serine 9 site increased in the hippocampus of the high dose group, compared with the model group. The phosphorylation level of GSK-3 beta at the tyrosine 216 site and PP2A were higher than that of the model group. There was no significant change in alpha protein level; IRS-1 at the serine 616 site, Akt at the serine 473 site, Tau protein at the serine 199396404422 site, and the phosphorylation level of the threonine 616 loci, while in the hippocampus of the low dose group, the phosphorylation level of Tau protein at the serine 404 and the threonine 231 loci was compared with the model group. The model group decreased. Akt decreased the phosphorylation level of serine 473 and GSK-3 beta at the serine 9 site. Conclusion: DNLA can reduce the learning and memory impairment and neuronal damage induced by STZ in rats. This effect is related to the DNLA improvement of STZ induced overphosphorylation of Tau protein in the hippocampus and the obstruction of the isdin signaling pathway in the hippocampus of rats.

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R285.5

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;某些因素對大鼠胃肌電的影響[J];基礎(chǔ)醫(yī)學(xué)與臨床;2001年S1期

2 陳同度,張昌穎;素食大鼠的貧血現(xiàn)象[J];營養(yǎng)學(xué)報(bào);1957年04期

3 陳偉強(qiáng);趙善廣;;自制注射用大鼠固定裝置[J];上海實(shí)驗(yàn)動(dòng)物科學(xué);1992年04期

4 肖柳英,林培英,馮昭明,張丹;不同周齡的SD大鼠生理、生化及體重的正常值測定[J];中藥新藥與臨床藥理;1996年03期

5 李淑云;簡易大鼠灌胃器的制作[J];錦州醫(yī)學(xué)院學(xué)報(bào);2001年04期

6 楊明智,陳積圣;一種大鼠抓取與固定的新工具介紹[J];上海實(shí)驗(yàn)動(dòng)物科學(xué);2001年03期

7 戴英,陸群;復(fù)方H_(505)對Wistar大鼠外周血的血液流變學(xué)指標(biāo)的影響[J];中國血液流變學(xué)雜志;2001年01期

8 韋應(yīng)波,孫喜慶,曹新生,姚永杰,馮岱雅,楊長斌;+Gz暴露時(shí)間對大鼠記憶功能和行為的影響[J];航天醫(yī)學(xué)與醫(yī)學(xué)工程;2003年01期

9 呂學(xué)軍,郭俊生,李敏,周利梅,張永娟;暈船大鼠體內(nèi)鐵含量的變化[J];中國職業(yè)醫(yī)學(xué);2003年04期

10 湯仁仙,王迎偉,王慧,周峰;201A中藥合劑對大鼠抗腎小球基底膜腎炎病變的影響[J];徐州醫(yī)學(xué)院學(xué)報(bào);2003年06期

相關(guān)會(huì)議論文 前10條

1 尹音;孫振宇;胡敏;李冬霞;;持續(xù)性高正加速度對大鼠顳頜關(guān)節(jié)損傷的作用[A];第八屆全國顳下頜關(guān)節(jié)病學(xué)及（牙合）學(xué)大會(huì)論文匯編[C];2011年

2 祝~=驤;iJ梊霞;洃克琴;崔素英;文允摪;，

本文編號：1815263