本文選題：犀角地黃湯 + 腦缺血再灌注　； 參考：《南京中醫(yī)藥大學(xué)》2017年碩士論文

【摘要】：目的:探討犀角地黃湯對(duì)局灶性腦缺血再灌注大鼠的腦保護(hù)作用及其對(duì)血管新生、神經(jīng)再生相關(guān)蛋白表達(dá)的影響,為犀角地黃湯治療腦梗死提供實(shí)驗(yàn)依據(jù)和藥理學(xué)基礎(chǔ)。方法:將健康雄性SPF級(jí)SD大鼠隨機(jī)分為5組:假手術(shù)組(Sham group),模型組(Model group),尼莫地平組(Nim group),犀角地黃湯高劑量組(XJDH-H group),犀角地黃湯低劑量組(XJDH-L group)。動(dòng)物在造模前1周開始每天給藥:犀角地黃湯高劑量組和低劑量組分別灌胃給予22.5g/kg·d和11.25g/kg·d的犀角地黃湯,尼莫地平組給予尼莫地平1mg/kg·d,假手術(shù)組和模型組給予等體積的生理鹽水,末次給藥2 h后進(jìn)行手術(shù)造模。采用線栓法阻塞大鼠中動(dòng)脈(middle cerebral artery occlusion,MCAO),建立局灶性腦缺血再灌注大鼠模型,造模2h后實(shí)現(xiàn)再灌注。24h大鼠自然清醒后,進(jìn)行神經(jīng)功能評(píng)分,HE染色觀察組織形態(tài)改變,TTC染色檢測(cè)腦梗死體積,采用Western Blot檢測(cè)大鼠缺血腦組織中 VEGF、bFGF、CD34、BDNF、GDNF 的表達(dá)。結(jié)果:1.犀角地黃湯對(duì)腦缺血再灌注大鼠神經(jīng)行為學(xué)的影響假手術(shù)組大鼠行為學(xué)無(wú)明顯改變,評(píng)分為0分;與假手術(shù)組比較,模型組大鼠神經(jīng)功能評(píng)分具有顯著性差異(p=0.000)。犀角地黃湯高劑量組神經(jīng)功能評(píng)分在第2天、第3天、第4天較模型組顯著降低(p0.01)。犀角地黃湯低劑量組神經(jīng)功能評(píng)分在第2天較模型組顯著降低(p0.01)。尼莫地平組神經(jīng)功能評(píng)分在第2天、第3天較模型組顯著降低(p0.01,p0.05)。2.犀角地黃湯對(duì)腦缺血再灌注損傷大鼠腦組織病理學(xué)的影響組織形態(tài)學(xué)結(jié)果顯示,假手術(shù)組的大鼠腦組織細(xì)胞形態(tài)規(guī)則,連接緊密。模型組神經(jīng)細(xì)胞結(jié)構(gòu)發(fā)生嚴(yán)重改變,形態(tài)結(jié)構(gòu)紊亂,大小不等,細(xì)胞核深染固縮,核仁、核膜不清,神經(jīng)細(xì)胞周圍間隙增寬。各給藥組可見(jiàn)神經(jīng)細(xì)胞缺血性改變均減輕。其中犀角地黃湯高劑量組神經(jīng)細(xì)胞缺血性改變明顯減輕。3.犀角地黃湯對(duì)大鼠腦缺血再灌注損傷腦梗死體積的影響假手術(shù)組的腦組織無(wú)梗死。與假手術(shù)組相比,模型組大鼠可見(jiàn)明顯的腦梗死。與模型組相比,犀角地黃湯高劑量組、尼莫地平組腦梗死體積明顯縮小(P0.01,P0.05),犀角地黃湯高劑量組效果優(yōu)于尼莫地平組(P0.05)。犀角地黃湯低劑量組腦梗死體積縮小不明顯(P0.05)。4.犀角地黃湯對(duì)腦缺血再灌注大鼠血管新生相關(guān)蛋白表達(dá)的影響在假手術(shù)組中,腦組織VEGF、bFGF、CD34的表達(dá)較低。模型組VEGF與bFGF的表達(dá)均顯著高于假手術(shù)組(p0.05),CD34表達(dá)無(wú)明顯差異(p0.05)。與模型組比較,犀角地黃湯低、高劑量組VEGF、bFGF、CD34的表達(dá)水平明顯升高(p0.05,p0.01),且呈一定的劑量依賴性。尼莫地平組與模型組比較也可顯著升高VEGF、bFGF、CD34的表達(dá)(p0.01)。5.犀角地黃湯對(duì)腦缺血再灌注大鼠神經(jīng)再生相關(guān)蛋白表達(dá)的影響與假手術(shù)組比較,模型組BDNF、GDNF表達(dá)升高不明顯(p0.05);與模型組比較,犀角地黃湯高劑量組和尼莫地平組BDNF、GDNF水平明顯升高(p0.01)。犀角地黃湯低劑量組BDNF水平明顯升高(p0.05),但GDNF升高不明顯(p0.05)。結(jié)論:犀角地黃湯可改善腦缺血再灌注后大腦的組織形態(tài)學(xué)變化,縮小腦梗死面積,對(duì)腦缺血再灌注損傷具有保護(hù)作用,其機(jī)制可能與上調(diào)VEGF、bFGF、CD34、BDNF、GDNF的表達(dá),促進(jìn)神經(jīng)和血管再生相關(guān)。
[Abstract]:Objective: To explore the protective effect of rhinoceros Dihuang Decoction on cerebral ischemia reperfusion in rats and its effect on angiogenesis and expression of nerve regeneration related protein, and provide experimental basis and pharmacological basis for the treatment of cerebral infarction by rhinoceros Dihuang Decoction. Methods: 5 healthy male SPF SD rats were randomly divided into 5 groups: sham operation group (Sham group), model Group (Model group), nimodipine group (Nim group), high dose group (XJDH-H group) of rhinoceros Dihuang Decoction (XJDH-H group) and low dose group (XJDH-L group) of rhinoceros Dihuang Decoction (XJDH-L group). The high dose group and low dose group of rhinoceros Dihuang Decoction group and low dose group were given the rhinoceros horn Rehmannia Soup for 22.5g/kg D and 11.25g/kg D, and nimodipine group was given to nimodipine group. The modipine 1mg/kg D, the sham operation group and the model group were given equal volume of physiological saline, and the last dose was 2 h after the operation. The rat middle artery (middle cerebral artery occlusion, MCAO) was blocked by the thread thrombus method. The rat model of focal cerebral ischemia reperfusion was established. After the model 2H was created, the reperfusion of.24h rats was naturally awake and nerve was carried out. HE staining was used to observe the changes of tissue morphology, TTC staining was used to detect the volume of cerebral infarction, and the expression of VEGF, bFGF, CD34, BDNF and GDNF in the ischemic brain tissue of rats was detected by Western Blot. Results: the neurobehavioral effects of 1. rhinoceros Dihuang Decoction on cerebral ischemia reperfusion rats were not significantly changed in the sham operation group, and the score was 0. The nerve function score of the model group was significantly different (p=0.000). The nerve function score in the high dose group of the rhinoceros Dihuang decoction was second days, third days and fourth days compared with the model group (P0.01). The nerve function score of the low dose group of rhinoceros Dihuang decoction was significantly lower than the model group (P0.01) in the second day. Nimodipine group's nerve function The score in second days and third days was significantly lower than that of the model group (P0.01, P0.05). The histopathological results of.2. rhinoceros Dihuang Decoction on cerebral ischemia reperfusion injury in rats showed that the cell morphology of the rat brain tissue in the sham operation group was regular and closely connected. The nuclear deep staining, nucleolus, nucleolus, nuclear membrane unclear, the peripheral space of the nerve cells widened. The ischemic changes of nerve cells in each group were reduced. The ischemic change of the nerve cell in the high dose group of rhinoceros Dihuang Decoction obviously alleviated the effect of.3. rhinoceros Dihuang Decoction on the volume of cerebral ischemia reperfusion injury in rats. Compared with the model group, the cerebral infarction volume in the nimodipine group was significantly reduced (P0.01, P0.05), and the effect of the high dose group of rhinoceros Dihuang decoction was better than that of the nimodipine group (P0.05). The effect of P0.05.4. rhinoceros Dihuang Decoction on the expression of angiogenesis related protein in cerebral ischemia reperfusion rats was lower in the sham operation group, and the expression of VEGF, bFGF and CD34 in the brain tissue was lower. The expression of VEGF and bFGF in the model group was significantly higher than that of the sham operation group (P0.05), and the expression of CD34 was not significantly different (P0.05). The expression level of VEGF, bFGF and CD34 increased significantly (P0.05, P0.01), and was dose-dependent. Nimodipine group and model group could also significantly increase VEGF, bFGF, CD34 expression (P0.01).5. (P0.01).5. rhino Dihuang Decoction on the expression of neural regeneration in cerebral ischemia reperfusion rats compared with the sham operation group, the model group BDNF, GDNF. The elevation of the expression was not obvious (P0.05). Compared with the model group, the level of BDNF and GDNF in the high dose group of rhinoceros Dihuang Decoction and the nimodipine group increased significantly (P0.01). The level of BDNF in the low dose group of rhinoceros Dihuang decoction was significantly increased (P0.05), but the increase of GDNF was not obvious (P0.05). Conclusion: rhinoceros Dihuang Decoction can improve the histomorphological changes of the brain after cerebral ischemia reperfusion. Reducing the area of cerebral infarction has protective effect on cerebral ischemia reperfusion injury. Its mechanism may be related to the up regulation of VEGF, bFGF, CD34, BDNF, GDNF, and the promotion of nerve and vascular regeneration.
【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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