Apelin-13對大鼠局灶性腦缺血—再灌注損傷的保護(hù)作用及其機(jī)制
發(fā)布時(shí)間：2019-02-16 12:24
【摘要】：目的：觀察Apelin-13對大鼠局灶性腦缺血-再灌注損傷的保護(hù)作用并探討其可能的機(jī)制 方法：采用線栓法建立S-D雄性大鼠局灶性缺血-再灌注損傷模型,缺血2小時(shí)再灌注72小時(shí)。進(jìn)行側(cè)腦室埋管,Apelin-13(0.1、1.0和10.0μg/kg)再灌注前15分鐘進(jìn)行側(cè)腦室注射。術(shù)后在不同時(shí)間點(diǎn)進(jìn)行神經(jīng)功能缺損評(píng)分。測量并計(jì)算大鼠腦組織含水量。采用2,3,5-氯化三苯基四氮唑(2,3,5-three phenyl tetrazolium chloride, TTC)染色觀察并計(jì)算腦梗死的體積和比例。H-E染色觀察大鼠腦組織切片的病理學(xué)改變。取缺血周邊腦組織,制備腦組織勻漿上清液,采用分光光度法檢測腦組織勻漿上清液中活性氧(reactive oxygen species, ROS)、丙二醛(malonaldehyde, MDA)含量和超氧化物歧化酶(superoxide dismutase, SOD)的活性；采用雙抗體夾心法測定腦組織勻漿上清液中總抗氧化力(total antioxidant capacity, T-AOC)的水平；采用ELISA法測定腦組織勻漿上清液中還原型谷胱甘肽(reduced glutathione hormon, GSH)的水平。采用實(shí)時(shí)定量PCR和Western blot檢測缺血周邊腦組織中腦源性神經(jīng)營養(yǎng)因子(brain derived neurotrophic factor, BDNF)和酪氨酸激酶B (tyrosine kinase B, TrkB) mRNA和蛋白的表達(dá)。 結(jié)果：(1)與缺血-再灌注損傷模型組比較,Apelin-13(1.0和10.0μg/kg)組大鼠神經(jīng)功能缺損明顯改善,肌力明顯增強(qiáng),神經(jīng)功能評(píng)分顯著性降低(均P0.05)。(2)與缺血-再灌注損傷模型組比較,Apelin-13(1.0和10.0μg/kg)處理顯著降低了腦組織的含水量、腦梗死體積和比例,呈現(xiàn)劑量依賴性(均P0.05)。(3)假手術(shù)組大鼠腦組織結(jié)構(gòu)正常,神經(jīng)元的細(xì)胞核居中,沒有神經(jīng)細(xì)胞的變性和壞死。而與假手術(shù)組比較,模型組大鼠腦組織中神經(jīng)細(xì)胞的數(shù)目明顯減少,可見明顯的組織水腫、神經(jīng)細(xì)胞變性和壞死,神經(jīng)元的細(xì)胞核出現(xiàn)不同程度的核固縮和溶解。與模型組相比,Apelin-13(1.0和10.0μg/kg)處理均可明顯改善缺血-再灌注后大鼠腦組織中神經(jīng)細(xì)胞的形態(tài),減輕核固縮和胞體腫脹程度,抑制神經(jīng)細(xì)胞的變性和壞死。(4)與缺血-再灌注損傷模型組比較,Apelin-13(1.0和10.0μg/kg)處理顯著降低了缺血周邊腦組織勻漿上清液中ROS水平和MDA含量(均P0.05),顯著增加了缺血周邊腦組織勻漿上清液中T-AOC水平、SOD的活性和GSH水平,呈現(xiàn)劑量依賴性(均P0.05)。(5)與缺血-再灌注損傷模型組比較,Apelin-13(1.0和10.0μg/kg)處理顯著增加了缺血周邊腦組織中BDNF和TrkB mRNA和蛋白的表達(dá),呈現(xiàn)劑量依賴性(P0.05)。 結(jié)論：Apelin-13對大鼠局灶性腦缺血-再灌注損傷有保護(hù)作用,其機(jī)制可能與Apelin-13抑制氧化應(yīng)激、上調(diào)BDNF及受體TrkB的表達(dá)有關(guān)。圖12幅,表1個(gè),參考文獻(xiàn)73篇。
[Abstract]:Objective: to observe the protective effect of Apelin-13 on focal cerebral ischemia-reperfusion injury in rats and explore its possible mechanism. Ischemia for 2 hours and reperfusion for 72 hours. Apelin-13 (0.1 渭 g/kg and 10.0 渭 g/kg) was injected 15 minutes before reperfusion. Neurological impairment scores were evaluated at different time points after operation. The water content of brain tissue was measured and calculated. The volume and proportion of cerebral infarction were observed and calculated by using the phenyl tetrazolium chloride, TTC) staining and H-E staining to observe the pathological changes of brain sections in rats. The supernatant of brain homogenate was prepared from the ischemic peripheral brain tissue. The content of reactive oxygen species (reactive oxygen species, ROS),) malondialdehyde (malonaldehyde, MDA) and the activity of superoxide dismutase (superoxide dismutase, SOD) in the supernatant of brain tissue homogenate were detected by spectrophotometry. The level of total antioxidant activity (total antioxidant capacity, T-AOC) in brain tissue homogenate supernatant was determined by double antibody sandwich method, and the level of reduced glutathione (reduced glutathione hormon, GSH) in brain tissue homogenate supernatant was measured by ELISA method. The expression of brain-derived neurotrophic factor (brain derived neurotrophic factor, BDNF) and tyrosine kinase (B (tyrosine kinase B, TrkB) mRNA) and protein in peripheral ischemic brain tissues were detected by real-time quantitative PCR and Western blot. Results: (1) compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) group significantly improved the nerve function defect and enhanced the muscle strength. Compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly decreased the water content of brain tissue, the volume and proportion of cerebral infarction, compared with the model group of ischemia-reperfusion injury (P0.05). (2). In a dose-dependent manner (P0.05). (3), the brain structure of the rats in the sham-operation group was normal, the nucleus of the neurons was in the middle, and there was no degeneration and necrosis of the neurons. Compared with the sham-operated group, the number of nerve cells in the brain tissue of the model group was significantly reduced, and obvious tissue edema, degeneration and necrosis of the neurons and pyknosis and dissolution of the nucleus of the neurons were observed in the model group. Compared with the model group, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly improved the morphology of neurons in the brain tissue after ischemia-reperfusion, and alleviated the degree of nuclear pyknosis and somatic swelling. Inhibition of degeneration and necrosis of nerve cells. (4) compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly decreased the level of ROS and MDA in the supernatant of ischemic brain tissue homogenate (P0.05), and significantly increased the T-AOC level in the supernatant of ischemic peripheral brain tissue homogenate. The activity of SOD and the level of GSH in a dose-dependent manner (P0.05). (5) were compared with those in the model group of ischemia-reperfusion injury. Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly increased the expression of BDNF, TrkB mRNA and protein in the ischemic peripheral brain tissue in a dose-dependent manner (P0.05). Conclusion: Apelin-13 has protective effect on focal cerebral ischemia-reperfusion injury in rats, and its mechanism may be related to the inhibition of oxidative stress and up-regulation of BDNF and receptor TrkB expression by Apelin-13. There are 12 figures, 1 table and 73 references.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3
本文編號(hào)：2424449
[Abstract]:Objective: to observe the protective effect of Apelin-13 on focal cerebral ischemia-reperfusion injury in rats and explore its possible mechanism. Ischemia for 2 hours and reperfusion for 72 hours. Apelin-13 (0.1 渭 g/kg and 10.0 渭 g/kg) was injected 15 minutes before reperfusion. Neurological impairment scores were evaluated at different time points after operation. The water content of brain tissue was measured and calculated. The volume and proportion of cerebral infarction were observed and calculated by using the phenyl tetrazolium chloride, TTC) staining and H-E staining to observe the pathological changes of brain sections in rats. The supernatant of brain homogenate was prepared from the ischemic peripheral brain tissue. The content of reactive oxygen species (reactive oxygen species, ROS),) malondialdehyde (malonaldehyde, MDA) and the activity of superoxide dismutase (superoxide dismutase, SOD) in the supernatant of brain tissue homogenate were detected by spectrophotometry. The level of total antioxidant activity (total antioxidant capacity, T-AOC) in brain tissue homogenate supernatant was determined by double antibody sandwich method, and the level of reduced glutathione (reduced glutathione hormon, GSH) in brain tissue homogenate supernatant was measured by ELISA method. The expression of brain-derived neurotrophic factor (brain derived neurotrophic factor, BDNF) and tyrosine kinase (B (tyrosine kinase B, TrkB) mRNA) and protein in peripheral ischemic brain tissues were detected by real-time quantitative PCR and Western blot. Results: (1) compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) group significantly improved the nerve function defect and enhanced the muscle strength. Compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly decreased the water content of brain tissue, the volume and proportion of cerebral infarction, compared with the model group of ischemia-reperfusion injury (P0.05). (2). In a dose-dependent manner (P0.05). (3), the brain structure of the rats in the sham-operation group was normal, the nucleus of the neurons was in the middle, and there was no degeneration and necrosis of the neurons. Compared with the sham-operated group, the number of nerve cells in the brain tissue of the model group was significantly reduced, and obvious tissue edema, degeneration and necrosis of the neurons and pyknosis and dissolution of the nucleus of the neurons were observed in the model group. Compared with the model group, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly improved the morphology of neurons in the brain tissue after ischemia-reperfusion, and alleviated the degree of nuclear pyknosis and somatic swelling. Inhibition of degeneration and necrosis of nerve cells. (4) compared with the model group of ischemia-reperfusion injury, Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly decreased the level of ROS and MDA in the supernatant of ischemic brain tissue homogenate (P0.05), and significantly increased the T-AOC level in the supernatant of ischemic peripheral brain tissue homogenate. The activity of SOD and the level of GSH in a dose-dependent manner (P0.05). (5) were compared with those in the model group of ischemia-reperfusion injury. Apelin-13 (1.0 渭 g/kg and 10.0 渭 g/kg) significantly increased the expression of BDNF, TrkB mRNA and protein in the ischemic peripheral brain tissue in a dose-dependent manner (P0.05). Conclusion: Apelin-13 has protective effect on focal cerebral ischemia-reperfusion injury in rats, and its mechanism may be related to the inhibition of oxidative stress and up-regulation of BDNF and receptor TrkB expression by Apelin-13. There are 12 figures, 1 table and 73 references.
【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3
【參考文獻(xiàn)】
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