【摘要】：目的觀察神經(jīng)肽Ghrelin在杏仁核電點(diǎn)燃大鼠外周血與腦組織中的變化，探討其原因及意義。 方法健康雄性SD大鼠64只，隨機(jī)分為點(diǎn)燃組（n=48）、假手術(shù)組（n=8）、空白組（n=8），點(diǎn)燃組以最后一次刺激為零點(diǎn)，再分為3h、6h、12h、24h、48h、72h六個(gè)亞組，每個(gè)亞組8只。點(diǎn)燃組植入電極后給予電刺激，制作杏仁核點(diǎn)燃膜型；假手術(shù)組只植入電極不予刺激；空白組不予處理。觀察大鼠行為學(xué)及腦電圖變化，采用ELISA、免疫組化和實(shí)時(shí)熒光定量PCR分別檢測血清中和下丘腦中辛�；疓hrelin表達(dá)情況。采用SPSS17.0統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析，計(jì)量資料用均數(shù)±標(biāo)準(zhǔn)差（x s）表示，，采用單因素方差分析，組間比較用最小顯著法（LSD-t）。 結(jié)果（1）杏仁核電點(diǎn)燃致癇大鼠癲癇發(fā)作情況：點(diǎn)燃大鼠在刺激最初表現(xiàn)為胡須抖動(dòng)、點(diǎn)頭，隨著刺激增多，出現(xiàn)抽搐、前肢陣攣等癥狀，完全點(diǎn)燃時(shí)表現(xiàn)為后肢站立并跌倒，全身抽動(dòng)，濕狗樣抖動(dòng)，發(fā)作時(shí)間約15s到數(shù)分鐘。點(diǎn)燃組后放閾為（441.54±52.43）μA，點(diǎn)燃速度為（7.12±2.13）d。實(shí)驗(yàn)組大鼠均可達(dá)到ⅴ級發(fā)作。 （2）杏仁核電點(diǎn)燃致癲癇大鼠血清中辛�；疓hrelin的濃度變化：空白組與假手術(shù)組之間差異無統(tǒng)計(jì)學(xué)意義（P0.05），各時(shí)間點(diǎn)辛酰基化Ghrelin水平與假手術(shù)組比較均降低（P0.05）。各時(shí)間點(diǎn)組間比較，3h組與6h組、6h組與12h組、24h組與48h組、48h組與72h組比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05），12h組與24h組比較差異無統(tǒng)計(jì)學(xué)意義（P0.05）。 （3）杏仁核電點(diǎn)燃致癇大鼠下丘腦中辛�；疓hrelin蛋白的表達(dá)：空白組與假手術(shù)組比較，差異無統(tǒng)計(jì)學(xué)意義（P0.05）；與假手術(shù)組比較，癲癇組各個(gè)時(shí)間點(diǎn)的辛�；疓hrelin表達(dá)水平均降低，各相鄰組間比較3h組與6h組、6h組與12h組、12h組與24h組、24h組與48h組、48h組與72h組間比較，差異有統(tǒng)計(jì)學(xué)意義（P0.05）。 （4）癲癇發(fā)作后大鼠下丘腦Ghrelin mRNA的表達(dá)：空白組與假手術(shù)組比較，GhrelinmRNA相對表達(dá)量差異無統(tǒng)計(jì)學(xué)意義（P0.05）。各時(shí)間點(diǎn)組與假手術(shù)組比較，GhrelinmRNA表達(dá)水平均降低，差異有統(tǒng)計(jì)學(xué)意義（P0.05）；各時(shí)間點(diǎn)組間比較，3h組與6h組、6h組與12h組、12h組與24h組、24h組與48h組間比較，差異有統(tǒng)計(jì)學(xué)意義平（P0.05）；48h組與72h組比較，差異無統(tǒng)計(jì)學(xué)意義（P0.05）。 結(jié)論癲癇發(fā)作后，血清與下丘腦中神經(jīng)肽Ghrelin水平均降低且變化趨勢是一致的，通過檢測血清中辛�；疓hrelin水平，可以反映下丘腦中相應(yīng)神經(jīng)肽的變化趨勢，可能成為臨床癲癇診斷和療效評估的一種新型檢測指標(biāo)。
[Abstract]:Objective to observe the changes of neuropeptide Ghrelin in peripheral blood and brain tissue of rats ignited by nuclear amygdala. Methods Sixty-four healthy male SD rats were randomly divided into six subgroups: kindling group (n = 48), sham operation group (n = 8) and blank group (n = 8). In the igniting group, electrical stimulation was given after electrode implantation, and amygdala kindling membrane was made; in sham-operation group, only electrode implantation was not stimulated; blank group was not treated. The changes of ethology and electroencephalogram of rats were observed. The expression of octanoylated Ghrelin in serum and hypothalamus was detected by ELISA, immunohistochemistry and real-time fluorescence quantitative PCR. The data were analyzed by SPSS17.0 software. The measurement data were expressed by mean 鹵standard deviation (x s), single factor analysis of variance (ANOVA) was used, and the least significant method (LSD-t) was used to compare the data between groups. Results (1) seizures of epileptic rats induced by nuclear amygdaloid kindling: the initial symptoms of epileptic rats induced by amygdaloid kindling were whiskers shaking, nodding, with the increase of stimuli, convulsions, clonus of forelimbs, and standing and falling of hind limbs when the rats were completely kindled. The whole body twitches, wet dog-like shake, attack time about 15 s to a few minutes. In the kindling group, the posterior release threshold was (441.54 鹵52.43) 渭 A, and the ignition speed was (7.12 鹵2.13) days. (2) the concentration of octanoylated Ghrelin in serum of epileptic rats induced by amygdaloid kindling: there was no significant difference between blank group and sham operation group (P0.05). Compared with sham operation group, the level of Ghrelin decreased (P0.05). The comparison was made between 3 h group and 6 h group, 6 h group and 12 h group, 24 h group and 48 h group, 48 h group and 72 h group. The difference was statistically significant (P0.05). There was no significant difference between 12h group and 24h group (P0.05). (3). The expression of octanoylated Ghrelin protein in hypothalamus of epileptic rats induced by nuclear amygdaloid kindling had no significant difference (P0.05). Compared with sham operation group, the expression of octanyl Ghrelin in epileptic group was decreased at all time points. The expression of octoyl Ghrelin was compared between 3 h group and 6 h group, 6 h group and 12 h group, 12 h group and 24 h group, 24 h group and 48 h group, 48 h group and 72 h group. The difference was statistically significant (P0.05). (4) the expression of Ghrelin mRNA in hypothalamus of rats after seizure: compared with sham operation group, there was no significant difference in the relative expression of GhrelinmRNA between blank group and sham operation group (P0.05). Compared with sham operation group, the expression of GhrelinmRNA in each time point group was significantly lower than that in sham operation group (P0.05), and the difference was statistically significant between 3h group and 6h group, 6h group and 12h group, 12h group and 24h group, 24h group and 48h group (P0.05). There was no significant difference between 48 h group and 72 h group (P0.05). Conclusion the levels of neuropeptide Ghrelin in serum and hypothalamus are decreased and the trend is the same after seizure. The level of octanoylated Ghrelin in serum can reflect the change trend of neuropeptide in hypothalamus. It may be a new detection index for the diagnosis and evaluation of clinical epilepsy.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R742.1

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