本文選題：PPARβ + NF-κB　； 參考：《重慶醫(yī)科大學》2014年博士論文

【摘要】：目的 通過測定大鼠蛛網膜下腔出血（SAH）后早期海馬過氧化物酶體增生物激活受體β（PPARβ），核因子-κB（NF-κB），基質金屬蛋白酶9（MMP-9）的表達變化，探討三者在蛛網膜下腔出血后參與早期腦損傷(early brain injury, EBI)的相關分子機制。 方法 1.建立大鼠SAH模型：采用視交叉前池注血法構建大鼠SAH模型。 2. PPARβ、NF-κB、MMP-9的表達：選取不同的時間點。 3. Real-time PCR方法檢測PPARβ、NF-κB、MMP-9mRNA表達，用Western blotting和免疫組化方法檢測PPARβ、NF-κB、MMP-9蛋白表達情況。 4.細胞凋亡的檢測：采用原位末端標記法（TUNEL）觀察大鼠海馬組織內神經元凋亡。 5.組織病理學檢測：通過檢測大鼠腦內伊文思藍（Evans blue, EB）含量觀察大鼠BBB通透性的改變，并通過檢測大鼠腦含水量（brainwater content, BWC）觀察大鼠腦水腫。 6.干預治療的作用：利用PPARβ特異性激動劑GW0742增加PPARβ的表達，觀察其與細胞凋亡、BBB通透性及腦水腫的變化關系以及對EBI的保護作用。 結果 1. PPARβ、NF-κB、MMP-9的表達：與sham組相比，大鼠SAH后6h海馬PPARβ蛋白表達和mRNA含量開始明顯減少，以后逐漸降低，72h達最低（P0.05)；與sham組相比，大鼠SAH后6h海馬NF-κB蛋白表達和mRNA含量開始明顯增加，以后逐漸增高，72h達高峰（P0.05)；與sham組相比，大鼠SAH后6h海馬MMP-9蛋白表達和mRNA含量開始明顯增加，以后逐漸增高，72h達高峰（P0.05)。 2. TUNEL檢測發(fā)現：大鼠SAH后6h開始出現TUNEL陽性細胞，隨后逐漸增多，72h時最多。SAH后6h，12h，24h，48h，72h大鼠TUNEL陽性細胞數均顯著高于sham組（P0.05）。 3.組織病理學檢測發(fā)現：SAH后12h大鼠腦內EB含量及BWC開始顯著升高，以后逐漸升高，72h達最高。與sham組相比，SAH后6h，12h，24h，48h，72h大鼠腦組織EB含量及BWC顯著增高（P0.05）。 4.干預治療顯示：SAH后72h，GW0742能夠明顯增加PPARβ的表達，同時降低NF-κB和MMP-9的表達（P0.05）；GWO742組海馬TUNEL陽性神經元數量與SAH組相比明顯增加（P0.05），GWO742組大鼠腦內EB含量和BWC與SAH組相比明顯降低（P0.05）。 結論 1.大鼠SAH后PPARβ可能通過NF-κB從轉錄水平調節(jié)MMP-9的表達，導致海馬神經元發(fā)生失巢性凋亡，參與早期腦損傷的病理過程。 2. PPARβ特異性激動劑GW0742能增加PPARβ的表達，減少細胞凋亡，減輕BBB通透性的改變，緩解腦水腫，具有潛在的神經保護作用。
[Abstract]:Objective to investigate the expression of peroxisome proliferator-activated receptor 尾 (PPAR 尾), nuclear factor- 魏 B (NF- 魏 B) and matrix metalloproteinase-9 (MMP-9) in rat hippocampus after subarachnoid hemorrhage (SAH). Objective: to explore the molecular mechanisms involved in early brain injury (early brain injury,) after subarachnoid hemorrhage. Method 1. Establishment of rat SAH model: the rat SAH model was established by injecting blood into the anterior cistern of optic chiasma. 2. Expression of PPAR 尾 -NF- 魏 B- MMP-9 at different time points. 3. 3%. Real-time PCR was used to detect the expression of MMP-9 mRNA in PPAR 尾 -NF- 魏 B, and Western blotting and immunohistochemistry were used to detect the expression of MMP-9 protein. 4. Detection of apoptosis: in situ end labeling (Tunel) was used to observe neuronal apoptosis in rat hippocampal tissue. Histopathological examination: the changes of BBB permeability and brain water content (brainwater content, BWC) were observed by detecting the content of Evans blue (EB) in rat brain. The effect of intervention therapy: GW0742, a specific agonist of PPAR 尾, was used to increase the expression of PPAR 尾, and to observe the relationship between the expression of PPAR 尾 and the permeability of apoptosis BBB and brain edema and the protective effect on EBI. Result 1. Compared with sham group, the expression of PPAR 尾 protein and mRNA began to decrease at 6 h after sham, and then decreased to the lowest at 72 h after SAH (P0.05), compared with sham group, NF- 魏 B protein expression and mRNA content in hippocampus began to increase at 6 h after sham. Compared with the sham group, the expression of MMP-9 protein and the content of MMP-9 in hippocampus began to increase significantly at 6 h after sham, and then increased to the peak at 72 h after SAH (P0.05). Tunel detection showed that the number of Tunel positive cells in rats was significantly higher than that in sham group (P0.05). The number of Tunel positive cells began to appear at 6 h after SAH and increased gradually at 72 h. The number of Tunel positive cells in rats was significantly higher than that in sham group at 12 h, 24 h and 48 h after SAH (P0.05). Histopathological examination showed that the content of EB and BWC in brain began to increase significantly at 12 h after WSAH and reached the highest level at 72 h later. Compared with sham group, the content of EB in brain tissue and the content of BWC in brain tissue of rats were significantly higher than those in sham group at 6 h, 12 h, 24 h and 48 h, respectively (P0.05). Intervention therapy showed that GW0742 significantly increased the expression of PPAR 尾 and decreased the expression of NF- 魏 B and MMP-9 (P0.05). The number of Tunel positive neurons in the hippocampus of the GWO 742 group was significantly higher than that in the SAH group (P0.05) and the content of EB and BWC in the brain of the GWO _ 742 group was significantly lower than that in the SAH group (P0.05). Conclusion 1. After SAH, PPAR 尾 may regulate the expression of MMP-9 through NF- 魏 B at the transcriptional level, resulting in apoptosis of hippocampal neurons, which may participate in the pathological process of early brain injury. 2. PPAR 尾 -specific agonist GW0742 can increase the expression of PPAR 尾, reduce apoptosis, alleviate the change of BBB permeability, relieve brain edema, and have potential neuroprotective effect.
【學位授予單位】：重慶醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R743.35

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相關期刊論文 前2條

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本文編號：2085177