本文選題：慢性氟中毒 + 神經(jīng)細胞��； 參考：《貴陽醫(yī)學院》2015年碩士論文

【摘要】：目的:通過檢測慢性氟中毒大鼠腦組織中神經(jīng)細胞凋亡率、凋亡誘導因子(apoptosis-inducing factor,AIF)和相關凋亡因子caspase在蛋白表達的水平,了解AIF介導的非caspase依賴性細胞凋亡途徑是否參與慢性氟中毒大鼠大腦神經(jīng)細胞凋亡過程,并探討其發(fā)生機制。方法:實驗用Sprague Dawley(SD)大鼠60只,雌雄各半,隨機分為兩組,即正常對照組:自由飲用自來水,飲水中含氟量0.5 mg/L;染氟組:飲水中加入氟化鈉,使氟含量為50 mg/L。兩組均飼以標準飼料(氟含量小于0.5mg/kg),實驗期為10個月。實驗結束時,觀察大鼠氟斑牙發(fā)生情況;用氟離子選擇電極法測定大鼠尿氟、骨氟含量;HE染色觀察大鼠大腦神經(jīng)組織學改變,尼氏染色觀察大鼠腦組織神經(jīng)細胞內尼氏小體形態(tài)學改變;免疫組化檢測大腦皮質及海馬AIF分布;蛋白印跡(Western blotting)檢測大腦皮質及海馬AIF、caspase-3及caspase-9的蛋白表達水平;流式細胞術(flow cytometr,FCM)檢測大鼠大腦皮質及海馬細胞凋亡率。結果:染氟成年大鼠出現(xiàn)不同程度的氟斑牙,尿氟及骨氟含量明顯高于對照組(P0.05);HE染色病理形態(tài)學檢查顯示,各組大鼠腦組織未見明顯異常改變,尼氏染色發(fā)現(xiàn)染氟組大鼠腦組織神經(jīng)細胞中尼氏體數(shù)量較對照組顏色變淺,數(shù)量減少;免疫組化結果顯示,氟中毒組海馬及皮質頂葉神經(jīng)細胞AIF陽性表達率與染色程度明顯高于對照組,差異有統(tǒng)計學意義(P0.05),但在額葉和枕葉差異無統(tǒng)計學意義(P0.05);蛋白印跡結果顯示,染氟組大鼠腦組織神經(jīng)細胞線粒體內AIF(57-k Da)的蛋白表達水平明顯低于對照組(P0.01),而細胞核中AIF的蛋白表達水平明顯高于對照組(P0.01);染氟組大鼠腦組織神經(jīng)細胞含半胱氨酸的天冬氨酸蛋白水解酶(cysteinyl aspartate specific proteinase,Caspase)-3的蛋白表達水平明顯高于對照組(P0.01),caspase-9的蛋白表達水平高于對照組(P0.05);流式細胞術結果顯示,染氟組大鼠海馬和皮質神經(jīng)細胞凋亡率與對照組相比顯著增高,差異有統(tǒng)計學意義(P0.05)。結論:以AIF為主的非caspase依賴性凋亡途徑與通過激活caspase介導的經(jīng)典途徑共同參與慢性氟中毒大鼠腦組織神經(jīng)細胞凋亡,是慢性氟中毒引起的腦損傷發(fā)生機制之一。
[Abstract]:Objective: to investigate the expression of apoptosis-inducing factor (AIFF) and apoptosis-inducing factor (caspase) in the brain of rats with chronic fluorosis. Objective: to investigate whether AIF mediated caspase dependent cell apoptosis is involved in the process of neurocyte apoptosis in rats with chronic fluorosis and to explore its mechanism. Methods: sixty male and female Sprague Dawley SD rats were randomly divided into two groups: normal control group: drinking tap water freely, fluorine content in drinking water 0.5 mg / L, fluoride group: adding sodium fluoride into drinking water, making fluorine content 50 mg / L. Both groups were fed with standard diet (fluoride content less than 0.5 mg / kg 路kg ~ (-1) for 10 months. At the end of the experiment, the occurrence of dental fluorosis in rats was observed, the urine fluoride was determined by fluorine ion selective electrode method, and the histological changes of brain nerve were observed by bone fluorine content and HE staining. Nissl's staining was used to observe the morphological changes of neuronal Nissl corpuscles in rat brain, the distribution of AIF in cerebral cortex and hippocampus was detected by immunohistochemistry, and the expression of AIF-caspase-3 and caspase-9 in cerebral cortex and hippocampus were detected by Western blotting (Western blotting). Apoptosis rate of cerebral cortex and hippocampal cells in rats was detected by flow cytometry (FCM). Results: dental fluorosis of different degrees was found in adult rats exposed to fluoride. The contents of urine fluoride and bone fluorine in adult rats were significantly higher than those in control group (P 0.05) and HE staining showed that there were no obvious abnormal changes in brain tissue of rats in each group. Nissl staining showed that the number of Nissl bodies in brain tissue of fluoride group was lighter than that of control group. The positive expression rate and staining degree of AIF in hippocampus and parietal cortex in fluorosis group were significantly higher than those in control group (P 0.05), but there was no significant difference in frontal lobe and occipital lobe (P 0.05). The expression level of AIF(57-k Daa in brain mitochondria of rats exposed to fluoride was significantly lower than that of control group (P 0.01), while the expression of AIF protein in nucleus was significantly higher than that in control group, and cysteine was found in nerve cells of rats exposed to fluoride. The protein expression level of cysteinyl aspartate specific proteinase Caspase- 3 was significantly higher than that of control group (P 0.01), and the expression of caspase-9 was significantly higher than that of control group (P 0.05), and the results of flow cytometry showed that the expression of caspase-9 was significantly higher than that of control group (P < 0.05), and the expression of caspase-9 was significantly higher than that of control group. The apoptosis rate of hippocampal and cortical neurons in fluoride group was significantly higher than that in control group (P 0.05). Conclusion: the AIF independent caspase dependent apoptosis pathway and the classical pathway mediated by caspase are involved in the neuronal apoptosis in the brain tissue of rats with chronic fluorosis, which is one of the mechanisms of brain injury induced by chronic fluorosis.
【學位授予單位】：貴陽醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R599.1

【參考文獻】

相關期刊論文 前4條

1 梅梅;于燕妮;陳楊;呂學霞;官志忠;;不同濃度氟對大鼠肝、腎功能及氧化應激的影響[J];貴陽醫(yī)學院學報;2007年04期

2 張麗英;單寅鑫;聶唯天;張歌;單春華;;AIF的生物特性與功能[J];現(xiàn)代生物醫(yī)學進展;2012年03期

3 趙艷;劉開泰;薛茜;;我國飲水型氟中毒預防措施效果評價[J];中國地方病學雜志;2006年02期

4 陳楊;于燕妮;呂學霞;官志忠;張華;;氧化應激在燃煤污染型氟中毒大鼠中的作用[J];中國實用醫(yī)藥;2008年04期

，

本文編號：1924954