本文選題：人參皂苷Rb1 + 癲癇��； 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：探討人參皂苷Rb1對(duì)青霉素誘導(dǎo)癲癇大鼠海馬神經(jīng)元凋亡和β-catenin蛋白表達(dá)的影響。方法：雄性Sprague-Dawley(SD)大鼠30只（體重250±50g），隨機(jī)分為4組：空白對(duì)照(BC)組（n=6）、人參皂苷Rb1對(duì)照(GC)組（n=8）、青霉素致癇(PE)組（n=8）和人參皂苷Rb1干預(yù)致癇(GPE)組（n=8）。人參皂苷Rb1對(duì)照組和人參皂苷Rb1干預(yù)致癇組大鼠在模型制備前1h腹腔注射1%人參皂苷Rb1注射液3ml/Kg (30mg/Kg)，空白對(duì)照組和青霉素致癇組大鼠則腹腔注射等劑量生理鹽水。大鼠癲癇模型的制備采用腹腔注射青霉素760萬(wàn)U/Kg的方法，即青霉素致癇組和人參皂苷Rb1干預(yù)致癇組大鼠腹腔注射80萬(wàn)U/ml青霉素注射液9.5ml/Kg，空白對(duì)照組和人參皂苷Rb1對(duì)照組大鼠則腹腔注射等劑量的生理鹽水。模型制備后觀察大鼠行為學(xué)改變，并記錄癲癇發(fā)作等級(jí)。模型制備后3h時(shí)，各組大鼠腹腔注射3％戊巴比妥鈉1ml/Kg麻醉，開胸暴露心臟，剪開右心房，將穿刺針經(jīng)左心室刺入升主動(dòng)脈，用生理鹽水快速灌注沖洗，然后用4%多聚甲醛500ml灌注固定，開顱，取出腦組織浸泡于4%多聚甲醛8～12h后固定。常規(guī)酒精脫水，石蠟包埋，連續(xù)切片（厚度4μm）。蘇木精-伊紅染色法（HE）染色，光鏡下觀察海馬神經(jīng)元組織學(xué)結(jié)構(gòu)變化；脫氧核糖核苷酸末端轉(zhuǎn)移酶介導(dǎo)的缺口末端標(biāo)記（TUNEL）法測(cè)海馬神經(jīng)元凋亡；免疫組化法測(cè)海馬神經(jīng)元β-catenin蛋白的陽(yáng)性表達(dá)。結(jié)果：⑴大鼠癲癇發(fā)作的行為學(xué)改變：空白對(duì)照組和人參皂苷Rb1對(duì)照組大鼠無(wú)癇樣發(fā)作表現(xiàn)；青霉素致癇組大鼠癲癇發(fā)作明顯，發(fā)作等級(jí)達(dá)Ⅲ級(jí)及以上，其中Ⅲ級(jí)3只(3/8), Ⅳ級(jí)2只(2/8)，Ⅴ級(jí)3只(3/8)；人參皂苷Rb1干預(yù)致癇組大鼠癲癇發(fā)作程度較青霉素致癇組明顯減輕，發(fā)作等級(jí)僅為Ⅰ-Ⅱ級(jí)，，其中Ⅰ級(jí)3只(3/8)，Ⅱ級(jí)5只(5/8)。⑵海馬神經(jīng)元組織結(jié)構(gòu)改變：空白對(duì)照組和人參皂苷Rb1對(duì)照組大鼠海馬神經(jīng)元形態(tài)結(jié)構(gòu)正常；青霉素致癇組和人參皂苷Rb1干預(yù)致癇組大鼠海馬神經(jīng)元出現(xiàn)變性和壞死損傷，但后者損傷程度較輕。(3)海馬神經(jīng)元細(xì)胞凋亡變化：各組大鼠海馬CA1、CA2、CA3和齒狀回區(qū)均有神經(jīng)元凋亡�？瞻讓�(duì)照組、人參皂苷Rb1對(duì)照組、青霉素致癇組和人參皂苷Rb1干預(yù)致癇組大鼠海馬CA1區(qū)凋亡神經(jīng)元計(jì)數(shù)（個(gè)）分別是2.5±1.05、1.75±0.71、3.5±0.93和1.63±0.74；CA2區(qū)凋亡神經(jīng)元計(jì)數(shù)分別是3.33±1.03、1.50±0.76、6.13±1.23、2.75±0.71； CA3區(qū)凋亡神經(jīng)元計(jì)數(shù)分別是3.83±1.17、3.75±0.71、8.38±1.3、5.38±1.19；齒狀回凋亡神經(jīng)元計(jì)數(shù)分別是3.5±1.38、3.38±0.92、21.75±3.85、10.88±3.36。與空白對(duì)照組相比，青霉素致癇組和人參皂苷Rb1干預(yù)致癇組大鼠海馬區(qū)凋亡陽(yáng)性細(xì)胞數(shù)明顯增高，差異具有統(tǒng)計(jì)學(xué)意義（P0.05）。與青霉素致癇組比較，人參皂苷Rb1干預(yù)致癇組海馬區(qū)凋亡陽(yáng)性細(xì)胞數(shù)明顯減少（P0.05)。與人參皂苷Rb1對(duì)照組比較，人參皂苷Rb1干預(yù)致癇組海馬區(qū)凋亡陽(yáng)性細(xì)胞數(shù)明顯增高（P0.05)。(4)海馬神經(jīng)元β-catenin蛋白表達(dá)變化：空白對(duì)照組、人參皂苷Rb1對(duì)照組、青霉素致癇組和人參皂苷Rb1干預(yù)致癇組大鼠海馬β-catenin蛋白表達(dá)OD值的平均值（AO）分別是(16.94±0.17)×10-2、(20.39±2.64)×10-2、(18.79±0.67)×10-2和(17.07±0.5)×10-2。與空白對(duì)照組相比，人參皂苷Rb1對(duì)照組和青霉素致癇組β-catenin蛋白陽(yáng)性表達(dá)高于空白對(duì)照組（P0.05）；與青霉素致癇組相比，人參皂苷Rb1干預(yù)致癇組β-catenin蛋白陽(yáng)性表達(dá)明顯減低（P0.05）；與人參皂苷Rb1對(duì)照組比較，人參皂苷Rb1干預(yù)致癇組β-catenin蛋白陽(yáng)性表達(dá)明顯減低（P0.05），差異均具有統(tǒng)計(jì)學(xué)意義。結(jié)論：青霉素誘導(dǎo)癲癇大鼠海馬神經(jīng)元凋亡增加和β-catenin蛋白表達(dá)升高；人參皂苷Rb1能夠減輕青霉素致癇大鼠的癲癇發(fā)作、減少海馬神經(jīng)元細(xì)胞凋亡和抑制β-catenin蛋白表達(dá)的上調(diào)。
[Abstract]:Objective: To investigate the effect of ginsenoside Rb1 on the apoptosis and the expression of beta -catenin protein in the hippocampus of penicillin induced epileptic rats. Methods: 30 male Sprague-Dawley (SD) rats (weight 250 + 50g) were randomly divided into 4 groups: blank control (BC) group (n=6), ginsenoside Rb1 control (n=8), penicillin induced epilepsy (PE) group (n=8) and ginsenoside 1 intervention induced epilepsy (GPE) group (n=8). Ginsenoside Rb1 control group and ginsenoside Rb1 intervention induced epileptic group rats before the model preparation of 1H intraperitoneal injection of 1% ginsenoside Rb1 injection 3ml/Kg (30mg/Kg), the blank control group and penicillin induced epilepsy rats were intraperitoneally injected with equal dose of physiological salt water. The preparation of the rat model of epilepsy was injected penicillin 7 in the abdominal cavity. 600 thousand U/Kg method, i. e. penicillin induced epilepsy group and ginsenoside Rb1 intervention, rats were intraperitoneally injected with 800 thousand U/ml penicillin injection 9.5ml/Kg, the blank control group and the ginsenoside Rb1 control group were intraperitoneally injected with equal dose of saline. After the model preparation, the rat behavior changes were observed and the seizure grade was recorded. Model preparation was made. After 3h, the rats were injected with 3% pentobarbital sodium 1ml/Kg anaesthesia, open the heart to expose the heart, cut open the right atrium, puncture the needle through the left ventricle into the ascending aorta and rinse with the saline rapid perfusion, then use 4% polyoxymethylene 500ml perfusion fixation, craniotomy, and remove the brain tissue after 4% polyformaldehyde 8 ~ 12h fixation. Conventional alcohol dehydration, Paraffin embedded, continuous slice (thickness of 4 m), hematoxylin eosin staining (HE) staining, observation of histological structure of hippocampal neurons under light microscope, apoptosis of hippocampal neurons by deoxyribonucleotide terminal transferase mediated nick end labeling (TUNEL), and immunohistochemical method to detect the positive expression of beta -catenin protein in hippocampal neurons. The behavioral changes of epileptic seizures in rats: there was no epileptic seizure in the blank control group and the ginsenoside Rb1 control group; the epileptic seizures in the group of penicillin induced epileptic rats were obvious, the level of the seizures reached grade III and above, of which 3 levels (3/8), grade IV (2/8), grade V 3 (3/8), and ginsenoside Rb1 intervention in epileptic group induced epileptic seizures in rats Compared with penicillin induced epilepsy group, the level of seizures was only grade I - II, including grade I 3 (3/8) and grade II 5 (5/8). (2) hippocampal neuron structure change: blank control group and ginsenoside Rb1 control group were normal in hippocampal neurons; penicillin induced epilepsy group and ginsenoside Rb1 intervention in epileptic group rats hippocampus neurons Degeneration and necrosis were observed, but the damage degree of the latter was mild. (3) apoptosis of hippocampal neurons: neuron apoptosis in hippocampus CA1, CA2, CA3 and dentate gyrus of rats in each group. The blank control group, the ginsenoside Rb1 control group, the penicillin induced epilepsy group and the Rb1 dry preeclampsia group of ginsenoside Rb1 rats' hippocampal CA1 region apoptotic neuron count () The number of apoptotic neurons in the CA2 region was 3.33 + 1.03,1.50 + 0.76,6.13 + 1.23,2.75 + 0.71, respectively, 2.5 + 1.05,1.75 + 0.71,3.5 + 0.93 and 1.63 + 0.74 respectively. The apoptotic neuron count in CA3 region was 3.83 + 1.17,3.75 + 0.71,8.38 + 1.3,5.38 + 1.19, and the number of apoptotic neurons in the dentate gyrus was 3.5 +. Compared with the blank control group, the number of apoptotic cells in hippocampus of epilepsy group induced by penicillin and ginsenoside Rb1 was significantly higher than that in epilepsy group (P0.05). Compared with penicillin induced epilepsy group, the number of apoptotic cells in hippocampus of epilepsy group induced by ginsenoside Rb1 was significantly decreased (P0.05). The control group of ginsenoside Rb1 was compared with the group of ginsenoside Rb1 (P0.05). The number of apoptotic cells in hippocampus of epileptic group was significantly increased by ginsenoside Rb1 (P0.05). (4) the changes in the expression of beta -catenin protein in hippocampal neurons: blank control group, ginsenoside Rb1 control group, penicillin induced epilepsy group and ginsenoside Rb1 intervention in epileptic group rats hippocampus beta -catenin protein expression o value (AO) was (16.94) + 0.17) * 10-2, (20.39 + 2.64) x 10-2, (18.79 + 0.67) x 10-2 and (17.07 + 0.5) x 10-2. compared with the blank control group, the positive expression of beta -catenin protein in the ginsenoside Rb1 control group and penicillin induced epilepsy group was higher than that in the blank control group (P0.05). Compared with the penicillin induced epilepsy group, the positive expression of beta -catenin protein in the epileptic group induced by ginsenoside Rb1 was significantly reduced. Lower (P0.05); compared with the ginsenoside Rb1 control group, the positive expression of beta -catenin protein in the epileptic group induced by ginsenoside Rb1 was significantly decreased (P0.05), and the difference was statistically significant. Conclusion: the apoptosis of hippocampus neurons in the epileptic rats induced by penicillin was increased and the beta -catenin protein table was increased, and ginsenoside Rb1 could reduce the large amount of penicillin induced epilepsy. Seizures in rats reduce the apoptosis of hippocampal neurons and inhibit the up regulation of the expression of beta -catenin protein.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R742.1

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 邢建華,仲崇斐,王曉潔;人參皂苷對(duì)胃腸腫瘤敏感性測(cè)定的實(shí)驗(yàn)研究[J];中國(guó)藥學(xué)雜志;2000年04期

2 初敏,許傳蓮,王秀全,丁立文;人參皂苷提取生產(chǎn)線設(shè)計(jì)[J];中藥研究與信息;2003年01期

3 李洪剛 ,楊義 ,何克江 ,欒宏偉 ,楊凌;分組和單體人參皂苷的批量制備[J];首都醫(yī)藥;2004年14期

4 劉奕明;楊柳;曾星;鄧遠(yuǎn)輝;馮怡;梁偉雄;;液相色譜-質(zhì)譜聯(lián)用法測(cè)定人參皂苷Re在健康人血漿的濃度[J];中國(guó)臨床藥理學(xué)雜志;2005年05期

5 陳云波;張大鵬;馮梅;王奇;程淑意;梁偉雄;溫澤淮;;人參皂苷Rg1對(duì)Aβ_(25-35)誘導(dǎo)的神經(jīng)細(xì)胞核因子-κB活化的影響[J];中國(guó)藥理學(xué)通報(bào);2007年05期

6 胡瑜;陳浩凡;臧林泉;巫瑋;;人參皂苷Rb_3對(duì)大鼠局灶性腦缺血損傷的保護(hù)作用[J];廣東藥學(xué)院學(xué)報(bào);2008年06期

7 李娜;孫印石;孫彥君;李偉;鄭毅男;;人參皂苷次生代謝產(chǎn)物的合成研究[J];吉林中醫(yī)藥;2008年07期

8 趙自明;潘華山;馮毅

本文編號(hào)：2078566