【摘要】：目的:探討人參皂苷Rb1對(duì)自噬調(diào)節(jié)的作用,以期能尋找一種減輕腦缺血缺氧后腦組織損傷的中藥,進(jìn)而減輕腦缺血缺氧對(duì)機(jī)體帶來(lái)的影響。方法:將30只SD大鼠隨機(jī)分成6組,每組5只,分別為高劑量組、中劑量組、低劑量組、空白對(duì)照組、假手術(shù)組、未做處理組,通過(guò)永久性結(jié)扎雙側(cè)頸總動(dòng)脈模擬腦缺血缺氧模型,分別按照1Omg/kg、5mg/kg、2.5mg/kg的濃度在造模后1-28天對(duì)高劑量組、中劑量組、低劑量組進(jìn)行腹腔注射,每天一次,并對(duì)假手術(shù)組和空白對(duì)照組每天給予超純水0.5ml,未做處理組不進(jìn)行腹腔注射作為對(duì)照。同時(shí)1、分別在造模前、造模45天后、造模90天后對(duì)各組大鼠進(jìn)行為期10天的水迷宮實(shí)驗(yàn)(具體包括定位航行實(shí)驗(yàn)、工作記憶實(shí)驗(yàn))以通過(guò)觀察各組大鼠尋找平臺(tái)的功能來(lái)評(píng)估大鼠的學(xué)習(xí)記憶能力進(jìn)而評(píng)價(jià)其認(rèn)知功能。2、在最后一次水迷宮實(shí)驗(yàn)結(jié)束后對(duì)大鼠斷頭取腦,收集其額葉及海馬組織,①Western blot檢測(cè)大鼠腦組織里Caspase-3、LC3B的含量,從而了解大鼠腦組織的自噬水平。②將腦組織HE染色后在光學(xué)顯微鏡下觀察(1)腦:有沒(méi)有變性、壞死的皮質(zhì)神經(jīng)元,有沒(méi)有增生的膠質(zhì)細(xì)胞,髓質(zhì)有沒(méi)有出現(xiàn)水腫。(2)海馬:海馬各區(qū)域的椎體細(xì)胞有沒(méi)有變性及壞死等改變。同時(shí)對(duì)腦組織出現(xiàn)的各種以上改變進(jìn)行程度輕重的評(píng)分。結(jié)果:①與正常對(duì)照組、假手術(shù)組初次水迷宮后即獲得學(xué)習(xí)記憶能力,在第45天及90天后尋找平臺(tái)時(shí)間明顯縮短相比,模型組大鼠出現(xiàn)明顯認(rèn)知功能障礙,具體表現(xiàn)為不管是工作記憶還是定位航行,模型組大鼠尋找平臺(tái)的持續(xù)時(shí)間及潛伏期均延長(zhǎng)(P0.05),上臺(tái)前搜索距離明顯延長(zhǎng)(P0.001),這一表現(xiàn)在造模90天后表現(xiàn)更顯著。而人參皂苷Rb1可降低腦缺血缺氧大鼠尋找平臺(tái)的持續(xù)時(shí)間及潛伏期(P0.05),同時(shí)明顯縮短模型大鼠上臺(tái)前的搜索距離(P0.001),濃度越高效果越明顯。②所有腦缺血缺氧模型組的Caspase-3含量增高(P0.05),LC3B含量降低(P0.05),人參皂苷Rb1作用于模型鼠后可抑制Caspase-3的表達(dá)同時(shí)促進(jìn)LC3B的表達(dá),且濃度越高效果越明顯。③HE染色后觀察模型組大鼠腦組織后可見:腦組織結(jié)構(gòu)與未做處理組相比,大多數(shù)區(qū)域皮層結(jié)構(gòu)消失,神經(jīng)元壞死,局部組織因高度疏松而表現(xiàn)為網(wǎng)狀,可以看到血管及膠質(zhì)細(xì)胞增殖;1區(qū)及2區(qū)海馬結(jié)構(gòu)不清楚,大多數(shù)錐體細(xì)胞壞死、消失,殘留的個(gè)別壞死細(xì)胞結(jié)構(gòu)不明;3區(qū)大多錐體細(xì)胞形狀不規(guī)則、核固縮、體積變性;病灶改變程度評(píng)分顯示模型組大鼠腦組織出現(xiàn)極重度改變;高劑量組、中劑量組腦組織為輕度改變,低劑量組腦組織為中度改變,空白劑量組腦組織為極重度改變,假手術(shù)組腦組織為輕微改變。表明人參皂苷Rb1可改善模型組大鼠的腦組織壞死能力且濃度越高改善作用越明顯。結(jié)論:人參皂苷Rb1可通過(guò)改善腦缺血缺氧模型組大鼠的學(xué)習(xí)記憶能力、調(diào)節(jié)自噬因子的表達(dá)、減輕腦組織壞死等作用降低腦缺血缺氧大鼠的腦組織損傷程度。
[Abstract]:Objective: To explore the effect of ginseng soap Rb1 on autophagy, in order to find a traditional Chinese medicine for alleviating brain injury after cerebral ischemia and anoxia, and to reduce the effects of cerebral ischemia and hypoxia on organism. Methods: Thirty SD rats were randomly divided into 6 groups, 5 rats in each group, the high dose group, the middle dose group, the low dose group, the blank control group, the sham operation group and the untreated group. 2. The concentration of 5mg/ kg was injected intraperitoneally in high dose group, medium dosage group and low dose group for 1 to 28 days after the model molding, and 0.5ml of ultrapure water was administered to the sham operation group and the blank control group every day, and no abdominal cavity injection was performed in the untreated group as the control group. At the same time, after the model was established for 45 days, a 10-day water maze test was carried out on each group of rats (including positioning navigation experiment) after 90 days respectively. working memory experiment) to evaluate the learning and memory ability of rats by observing the function of each group of rat searching platform to evaluate its cognitive function. Western blot was used to detect the level of Caspase-3 and LC3B in brain tissue of rats. He stained the brain tissue with HE and observed (1) the brain under an optical microscope: whether there was degeneration, necrosis of the cortical neurons, the presence of proliferating glial cells, and the presence of edema in the medulla. (2) hippocampus: there is no change in the degeneration and necrosis of the vertebral body cells in each region of the hippocampus. At the same time, the severity of the changes in brain tissue was scored. Results: Compared with the normal control group and the sham operation group, after the initial water maze, the learning and memory ability was acquired. After 45 days and 90 days, there was obvious cognitive dysfunction in the model group. The duration and latency of search platform for model group were prolonged (P0.05). The duration and incubation period (P <0.01) of the model rats were significantly shortened (P <0.01), and the higher the concentration was, the higher the concentration was. The expression of Caspase-3 was decreased (P0.05). The expression of Caspase-3 was inhibited at the same time, and the higher the concentration was. After HE staining, it was found that the structure of the brain tissue disappeared, the neurons were necrotic, the local tissue was reticulate because of the high degree of osteoporosis, and the proliferation of vascular and glial cells could be seen. In 1 and 2 regions, the structure of hippocampal formation was not clear, most of the pyramidal cells were necrotic, disappeared, and the residual individual necrotic cells were unknown. In the high-dose group, the brain tissue of the middle-dose group was slightly changed, the brain tissue of the low-dose group was moderately changed, the brain tissue of the blank dose group was very severe, and the brain tissue of the sham-operated group was slightly changed. The results showed that Rb1 could improve the brain tissue necrosis and the higher the concentration. Conclusion: Ginseng soap Rb1 can improve the learning and memory ability of rats with cerebral ischemia and hypoxia model, regulate the expression of autophagy factor, reduce the brain tissue necrosis, and decrease the damage degree of brain tissue in rats with cerebral ischemia and hypoxia.
【學(xué)位授予單位】：南京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285.5

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