KLF4在PTZ點(diǎn)燃模型大鼠海馬苔蘚纖維出芽過程中的動(dòng)態(tài)表達(dá)
發(fā)布時(shí)間：2019-01-02 17:17
【摘要】：目的采用戊四氮(Pentamethazol, PTZ)點(diǎn)燃癲癇大鼠模型,觀察海馬中轉(zhuǎn)錄因子Kruppe樣因子4(Kruppel-like factor4, KLF4)的表達(dá)變化,結(jié)合海馬苔蘚纖維出芽(Mossy fiber sprouting, MFS)的動(dòng)態(tài)改變,探討其在癲癇發(fā)生過程中的可能作用。 方法清潔級(jí)健康成年雄性SD大鼠90只,隨機(jī)分為戊四氮實(shí)驗(yàn)組和生理鹽水對(duì)照組,其中戊四氮實(shí)驗(yàn)組60只,生理鹽水對(duì)照組30只。戊四氮組腹腔注射PTZ30mg/kg,每天一次,對(duì)照組同時(shí)注射等量生理鹽水。按第一次腹腔注射后3天、1周、2周、4周和6周在兩組內(nèi)隨機(jī)平均分為5個(gè)亞組,每個(gè)亞組再隨機(jī)平均分為兩個(gè)小組：一組用于免疫組化,檢測各時(shí)間點(diǎn)大鼠海馬中的KLF4表達(dá)；另一組用于Timm染色,檢測苔蘚纖維出芽情況。 結(jié)果 1.動(dòng)物模型 大鼠多在連續(xù)注射PTZ21～28天后被點(diǎn)燃,平均24.5±2.3天。模型點(diǎn)燃率為100%,死亡率為2.2%。實(shí)驗(yàn)組大鼠成功點(diǎn)燃后出現(xiàn)誘發(fā)發(fā)作及自發(fā)發(fā)作；對(duì)照組未出現(xiàn)任何癇性發(fā)作。 2. KLF4蛋白的表達(dá) 對(duì)照組大鼠海馬CA1、CA3、齒狀回區(qū)KLF4蛋白表達(dá)在3天后逐漸升高,至1周時(shí)達(dá)高峰,并持續(xù)至第6周；實(shí)驗(yàn)組大鼠海馬CA1、CA3、齒狀回區(qū)KLF4蛋白表達(dá)在第3天后未升高,而是保持在穩(wěn)定水平,持續(xù)至第6周。兩組間CA1區(qū)KLF4蛋白表達(dá)在3天、1周時(shí)有統(tǒng)計(jì)學(xué)差異；CA3區(qū)僅1周組有統(tǒng)計(jì)學(xué)差異；齒狀回區(qū)在1周和2周組有統(tǒng)計(jì)學(xué)差異；余各區(qū)各時(shí)間點(diǎn)兩組間無統(tǒng)計(jì)學(xué)差異。 3.苔蘚纖維出芽的動(dòng)態(tài)變化 實(shí)驗(yàn)組大鼠海馬CA3區(qū)苔蘚纖維出芽與對(duì)照組CA3區(qū)相比,從第3天開始實(shí)驗(yàn)組出現(xiàn)苔蘚纖維出芽,評(píng)分逐漸增加,至第2周開始兩組間出現(xiàn)統(tǒng)計(jì)學(xué)差異(P0.05),至第6周時(shí)達(dá)高峰。實(shí)驗(yàn)組大鼠海馬齒狀回區(qū)苔蘚纖維出芽與對(duì)照組齒狀回區(qū)相比,從第4周開始實(shí)驗(yàn)組出現(xiàn)苔蘚纖維出芽,評(píng)分逐漸增加,至第6周時(shí)達(dá)高峰并在兩組間出現(xiàn)統(tǒng)計(jì)學(xué)差異(P0.05)。對(duì)照組大鼠海馬CA3區(qū)苔蘚纖維出芽評(píng)分均僅0-1分,各時(shí)間點(diǎn)間無統(tǒng)計(jì)學(xué)差異；齒狀回區(qū)各時(shí)間點(diǎn)未發(fā)現(xiàn)苔蘚纖維出芽。 結(jié)論 在戊四氮點(diǎn)燃大鼠模型中,海馬KLF4蛋白早期表達(dá)下調(diào)可能促進(jìn)苔蘚纖維出芽。
[Abstract]:Objective to observe the expression of transcription factor Kruppe like factor 4 (Kruppel-like factor4, KLF4) in hippocampus of epileptic rats kindled with pentylenetetrazol (Pentamethazol, PTZ), and to observe the dynamic changes of (Mossy fiber sprouting, MFS) in mossy fibers. To explore the possible role of Epilepsy in the process of epilepsy. Methods 90 healthy male SD rats of clean grade were randomly divided into pentylenetetrazol group (n = 60) and saline control group (n = 30). Pentylenetetrazol group was intraperitoneally injected with PTZ30mg/kg, once a day, while the control group was injected with the same amount of normal saline at the same time. Three days, one week, two weeks, four weeks and six weeks after the first intraperitoneal injection, each subgroup was randomly divided into five subgroups, and each subgroup was randomly divided into two groups: one for immunohistochemistry. The expression of KLF4 in hippocampus of rats was detected at different time points. The other group was used for Timm staining to detect the sprouting of moss fibers. Result 1. Most of the animal model rats were kindled after continuous injection of PTZ21~28, with an average of 24.5 鹵2.3 days. The model ignition rate was 100 and the mortality was 2.2. The rats in the experimental group had induced seizures and spontaneous seizures after successful kindling, while in the control group there were no epileptic seizures. 2. The expression of KLF4 protein in hippocampal CA1,CA3, dentate gyrus of control group increased gradually after 3 days, reached its peak at 1 week and lasted until 6 weeks. The expression of KLF4 protein in the dentate gyrus of hippocampal CA1,CA3, did not increase after 3 days in the experimental group, but remained at a stable level until 6 weeks. The expression of KLF4 protein in CA1 region was significantly different between the two groups at 3 days and 1 week; in the CA3 region only 1 week group there was statistical difference; in dentate gyrus area at 1 week and 2 weeks group there was statistical difference; there was no statistical difference between the two groups at other time points. 3. Dynamic changes of mossy fiber sprouting in the experimental group compared with the control group, the mossy fiber sprouts appeared in the experimental group from the 3rd day, and the score increased gradually compared with the control group. At the beginning of the second week, there was a statistical difference between the two groups (P0.05), and reached the peak at the 6th week. Compared with the control group, the mossy fiber sprouts from the dentate gyrus of the experimental group and the control group, the mossy fiber sprouts appeared in the experimental group from the 4th week, and the score increased gradually. Reached the peak at the 6th week and there was statistical difference between the two groups (P0.05). In the control group, the mossy fiber sprouting scores in the hippocampal CA3 area were only 0-1, and there was no statistical difference between the different time points, and no mossy fiber sprouting was found in the dentate gyrus at each time point. Conclusion in pentylenetetrazol kindled rat model, early down-regulation of hippocampal KLF4 protein may promote mossy fiber sprouting.
【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】：2013
【分類號(hào)】：R742.1;R-332
本文編號(hào)：2398764
[Abstract]:Objective to observe the expression of transcription factor Kruppe like factor 4 (Kruppel-like factor4, KLF4) in hippocampus of epileptic rats kindled with pentylenetetrazol (Pentamethazol, PTZ), and to observe the dynamic changes of (Mossy fiber sprouting, MFS) in mossy fibers. To explore the possible role of Epilepsy in the process of epilepsy. Methods 90 healthy male SD rats of clean grade were randomly divided into pentylenetetrazol group (n = 60) and saline control group (n = 30). Pentylenetetrazol group was intraperitoneally injected with PTZ30mg/kg, once a day, while the control group was injected with the same amount of normal saline at the same time. Three days, one week, two weeks, four weeks and six weeks after the first intraperitoneal injection, each subgroup was randomly divided into five subgroups, and each subgroup was randomly divided into two groups: one for immunohistochemistry. The expression of KLF4 in hippocampus of rats was detected at different time points. The other group was used for Timm staining to detect the sprouting of moss fibers. Result 1. Most of the animal model rats were kindled after continuous injection of PTZ21~28, with an average of 24.5 鹵2.3 days. The model ignition rate was 100 and the mortality was 2.2. The rats in the experimental group had induced seizures and spontaneous seizures after successful kindling, while in the control group there were no epileptic seizures. 2. The expression of KLF4 protein in hippocampal CA1,CA3, dentate gyrus of control group increased gradually after 3 days, reached its peak at 1 week and lasted until 6 weeks. The expression of KLF4 protein in the dentate gyrus of hippocampal CA1,CA3, did not increase after 3 days in the experimental group, but remained at a stable level until 6 weeks. The expression of KLF4 protein in CA1 region was significantly different between the two groups at 3 days and 1 week; in the CA3 region only 1 week group there was statistical difference; in dentate gyrus area at 1 week and 2 weeks group there was statistical difference; there was no statistical difference between the two groups at other time points. 3. Dynamic changes of mossy fiber sprouting in the experimental group compared with the control group, the mossy fiber sprouts appeared in the experimental group from the 3rd day, and the score increased gradually compared with the control group. At the beginning of the second week, there was a statistical difference between the two groups (P0.05), and reached the peak at the 6th week. Compared with the control group, the mossy fiber sprouts from the dentate gyrus of the experimental group and the control group, the mossy fiber sprouts appeared in the experimental group from the 4th week, and the score increased gradually. Reached the peak at the 6th week and there was statistical difference between the two groups (P0.05). In the control group, the mossy fiber sprouting scores in the hippocampal CA3 area were only 0-1, and there was no statistical difference between the different time points, and no mossy fiber sprouting was found in the dentate gyrus at each time point. Conclusion in pentylenetetrazol kindled rat model, early down-regulation of hippocampal KLF4 protein may promote mossy fiber sprouting.
【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】：2013
【分類號(hào)】：R742.1;R-332
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相關(guān)期刊論文 前2條
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