本文關(guān)鍵詞： 視神經(jīng)切斷 電針 視網(wǎng)膜神經(jīng)節(jié)細胞 神經(jīng)元存活 谷氨酸 谷氨酸轉(zhuǎn)運體-1　出處：《第四軍醫(yī)大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的：研究不同穴位和頻率的電針刺激對成年大鼠視神經(jīng)切斷后視網(wǎng)膜神經(jīng)節(jié)細胞（節(jié)細胞）存活的作用，并探討電針刺激對受損節(jié)細胞的保護機制。 實驗一：隨機選取54只SD大鼠，切斷右側(cè)視神經(jīng)并用熒光金（FluoroGold, FG）逆行性標記節(jié)細胞后，隨機分為9組：對照組（含2、7和14d對照組）及不同穴位、頻率和傷后存活時間組合的電針組（含假穴4/20Hz電針7d組、百會穴4/20Hz電針7d組、睛明穴2Hz電針7d組、睛明穴4/20Hz電針2、7、14d組），每組6只動物。對應(yīng)時間點處死動物取右眼視網(wǎng)膜后計數(shù)被熒光金標記的節(jié)細胞并計算相應(yīng)的密度（細胞/mm2），數(shù)值用Mean±SEM表示。結(jié)果：（1）睛明穴4/20Hz電針7d組存活節(jié)細胞平均密度（1600±72）顯著高于對照（1104±41）、假穴（1222±68）和百會穴（1168±56）組（P0.01），而假穴和百會穴組與對照組間無顯著性差異；（2）睛明穴2Hz與睛明穴4/20Hz電針7d組節(jié)細胞密度雖無顯著性差異，但均顯著高于對照7d組（P0.01）；（3）分別以4/20Hz電針刺激睛明穴2、7、14d，與對照組相同時間點相比，僅在傷后7d出現(xiàn)節(jié)細胞密度的顯著增高（P0.01）。 實驗二：隨機選取60只大鼠并隨機分為10組，即正常組、假手術(shù)2、7、14d組（手術(shù)暴露右側(cè)視神經(jīng)但不切斷），，另外36只大鼠切斷右側(cè)視神經(jīng)后再隨機分為2、7、14d對照組和2、7、14d電針組（即2、7、14d睛明穴4/20Hz電針組），每組6只動物。于各組相應(yīng)時間點麻醉動物后取動物右眼收集房水，并利用高效液相色譜檢測其谷氨酸濃度（μmol/L），數(shù)值用Mean±SEM表示。結(jié)果：對照2、7、14d組（濃度分別為59.74±4.62、88.17±1.94、56.56±1.15）及電針2、7、14d組（濃度分別為47.62±4.29、54.72±1.98、52.27±1.16）房水谷氨酸濃度均顯著高于正常組及2、7、14d假手術(shù)組（濃度分別為43.54±0.53、43.71±0.66、43.21±0.94、43.04±0.44）（P0.01）；電針2、7d組房水谷氨酸濃度分別顯著低于對照組2、7d組（P0.01）；電針14d組房水谷氨酸濃度與對照14d組無差異。 實驗三：隨機選取54只大鼠切斷右側(cè)視神經(jīng)后，隨機分為6組：2、7、14d對照組和2、7、14d電針組（即2、7、14d睛明穴4/20Hz電針組），每組9只動物。分別利用免疫組織化學染色、Western-blot方法檢測大鼠視網(wǎng)膜中分別參與谷氨酸重吸收與代謝的谷氨酸轉(zhuǎn)運體-1和谷氨酰胺合成酶的表達。結(jié)果：電針7d組谷氨酸轉(zhuǎn)運體-1表達顯著高于對照7d組（P0.05）；對照2、14d組分別與電針2、14d組的谷氨酸轉(zhuǎn)運體-1表達無顯著差異。視神經(jīng)切斷后電針組和對照組谷氨酰胺合成酶表達沒有發(fā)生變化。 結(jié)論：切斷成年大鼠視神經(jīng)后每日以4/20Hz頻率的電針刺激睛明穴1次，可在傷后7d促進視網(wǎng)膜谷氨酸轉(zhuǎn)運體-1對傷側(cè)眼房水內(nèi)谷氨酸的重吸收，從而降低房水內(nèi)谷氨酸的含量，減輕谷氨酸興奮性毒性，延緩受損節(jié)細胞的死亡。
[Abstract]:Aim: to study the effects of electroacupuncture stimulation at different points and frequencies on the survival of retinal ganglion cells (ganglion cells) after optic nerve transection in adult rats and to explore the protective mechanism of electroacupuncture stimulation on injured ganglion cells. Experiment 1: Fifty-four Sprague-Dawley rats were randomly divided into 9 groups: control group (control group) and different acupoints after transection of the right optic nerve and fluorescent Fluoro Gold (FG) retrograde labeled ganglion cells. The frequency and survival time after injury were combined with electroacupuncture group (including 4 / 20 Hz electroacupuncture group for 7 days, Baihui point 4 / 20 Hz electroacupuncture group for 7 days, Jinming point 2 Hz electroacupuncture group for 7 days). There were 6 animals in each group after 4 / 20 Hz electroacupuncture for 14 days. The right eye retina was killed at the corresponding time point to count the fluorescent gold labeled ganglion cells and calculate the corresponding density (cell per mm2, expressed in Mean 鹵SEM. The result was 1: 1). The mean cell density of surviving ganglion cells in the electroacupuncture group was significantly higher than that in the control group (1104 鹵41g), the control group (1222 鹵68) and the Baihui acupoint group (1168 鹵56), but there was no significant difference between the sham acupoint group and Baihui acupoint group and the control group (P 0.01), although there was no significant difference in cell density between the Jingming acupoint (2Hz) and Jinming acupoint (4 / 20Hz) after 7 d electroacupuncture, although there was no significant difference in cell density between the sham acupoint and Baihui acupoint group and the control group. But it was significantly higher than that of the control group on the 7th day (P 0.01). Compared with the control group at the same time point, there was a significant increase in the cell density of the ganglion on the 7th day after injury (P 0.01) after 4 / 20 Hz electroacupuncture stimulation on the Jingming acupoint for 2 ~ 7 ~ 14 days, respectively, and compared with the control group at the same time point, there was a significant increase in the cell density. Experiment 2: 60 rats were randomly selected and randomly divided into 10 groups, that is, the normal group. The rats in the sham operation group (the right optic nerve was exposed without transection of the right optic nerve, and the other 36 rats were randomly divided into two groups: the control group, the control group, and the electroacupuncture group, which were treated with electroacupuncture on the 2nd day, 7d, 14d, respectively), each group had 6 animals in each group, and the other 36 rats were divided into two groups: the control group, the control group, and the electroacupuncture group, which had 4 / 20 Hz electroacupuncture on the right optic nerve. After anaesthetized animals were anesthetized, aqueous humor was collected from the right eye of the animals. The concentration of glutamic acid (渭 mol / L) was measured by high performance liquid chromatography (渭 mol / L) (Mean 鹵SEM). Results: the concentration of glutamate in the control group (59.74 鹵4.62n 88.17 鹵1.94 鹵56.56 鹵1.15) and electroacupuncture group (47.62 鹵4.2954.72 鹵1.98 鹵52.27 鹵1.16) were significantly higher than those in the control group and the control group for 714d. The concentration of glutamate in aqueous humor in the operation group (43.54 鹵0.53U 43.71 鹵0.66U 43.21 鹵0.94U 43.04 鹵0.44 P0.01A), in the electroacupuncture group of 2d 7d was significantly lower than that in the control group (P 0.01g), and there was no difference between the 14 d group and the control group (14d) in the concentration of glutamate in aqueous humor. Experiment 3: 54 rats were randomly selected after the right optic nerve was transected. The rats were randomly divided into 6 groups: control group (n = 6) and control group (n = 14) and electroacupuncture group (n = 9, n = 9 in each group). Immunohistochemical staining was used to detect glutamate reabsorption and metabolism in the retina of rats. Results: the expression of glutamate transporter-1 in electroacupuncture group was significantly higher than that in control group on 7 d group (P 0. 05), but there was no significant difference between control group and electroacupuncture group on day 214 and electroacupuncture group on day 214. the expression of glutamate transporter-1 in electroacupuncture group was significantly higher than that in control group on 7 d group (P < 0. 05). After optic nerve transection, the expression of glutamine synthase did not change in electroacupuncture group and control group. Conclusion: after transection of the optic nerve of adult rats, stimulation of Jinming point with 4 / 20Hz frequency electroacupuncture once a day can promote the reabsorption of glutamate transporter 1 in the injured eyes on the 7th day after injury, thereby reducing the content of glutamate in aqueous humor. Reduce glutamate excitotoxicity and delay the death of damaged ganglion cells.
【學位授予單位】：第四軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R245;R774.1

【參考文獻】

相關(guān)期刊論文 前3條

1 張志英,嚴振國;電針對脊髓損傷后一氧化氮合酶表達的影響[J];中國臨床康復(fù);2002年02期

2 勞寧,熊利澤,路志紅,劉艷紅,鞏固,雷毅;不同頻率電針預(yù)處理對誘導腦缺血耐受程度的差異[J];中國臨床康復(fù);2005年09期

3 費宏,謝國璽,韓濟生;LOW AND HIGH FREQUENCY ELECTROCU-PUNCTURE STIMULATIONS RELEASE (Met~5)ENKEPHALIN AND DYNORPHIN A IN RAT SPINAL CORD[J];Chinese Science Bulletin;1987年21期

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