【摘要】：目的:用線栓法復(fù)制大鼠腦缺血/再灌注(I/R)損傷模型,以細(xì)胞色素P450表氧化酶CYP-EETs信號(hào)通路為調(diào)控靶點(diǎn),研究可溶性表氧化物水解酶抑制劑(sEHi)TPPU對(duì)腦缺血/再灌注損傷所致血腦屏障(BBB)破壞的作用,探索腦缺血治療新靶點(diǎn)。方法:通過(guò)線栓法建立大鼠MCAO腦缺血/再灌注模型;大鼠隨機(jī)分為5組,sham組、MCAO組、TPPU(1.0mg/kg)組、TPPU(0.3mg/kg)組、TPPU(0.1mg/kg)組;激光多普勒血流儀及激光散斑評(píng)估模型成功與否,TTC法檢測(cè)MCAO梗死體積,伊文氏藍(lán)法檢測(cè)血腦屏障通透性,采用免疫熒光染色、western blot等方法檢測(cè)MCAO及TPPU干預(yù)后血腦屏障相關(guān)蛋白含量的變化,干濕重法評(píng)估腦水腫程度,Garcia18分制行為學(xué)評(píng)分法評(píng)估神經(jīng)功能缺損及恢復(fù)情況。結(jié)果:線栓法t-MCAO模型成功率高、死亡率低,激光多普勒血流儀能夠全程動(dòng)態(tài)反映MCA(middle cerebral artery大腦中動(dòng)脈)區(qū)域腦組織的血流變化,有助于判定MCA是否形成有效梗阻;激光散斑成像能夠直觀顯示感興趣區(qū)血管的血流變化,缺點(diǎn)是損傷大且不能連續(xù)不間斷的監(jiān)測(cè);MCAO后大鼠均出現(xiàn)神經(jīng)功能缺損癥狀,與sham組比較有顯著統(tǒng)計(jì)學(xué)差異(P0.01);MCAO大鼠血腦屏障通透性增高,梗死側(cè)腦組織可見(jiàn)大量伊文思藍(lán)滲漏,進(jìn)而繼發(fā)血管性腦水腫;TTC染色直觀反映出梗死面積,與神經(jīng)功能缺損嚴(yán)重程度相符合;與MCAO組比較,可溶性環(huán)氧化物水解酶抑制劑TPPU可以減小腦皮質(zhì)梗死體積(P0.05),而對(duì)紋狀體梗死作用不明顯(P0.05);TPPU能夠減輕MCAO后血腦屏障緊密連接(tight junction,TJ)蛋白的丟失(P0.05),有助于維持血腦屏障完整性,減少伊文思藍(lán)向腦組織滲漏(P0.001),進(jìn)而減輕血管性腦水腫(P0.05);TPPU能夠改善MCAO大鼠腦梗死后神經(jīng)功能缺損癥狀,各劑量組與對(duì)照組比較均有統(tǒng)計(jì)學(xué)意義(P0.01);上述獲益隨TPPU劑量增大而增加,呈劑量依賴性。結(jié)論:可溶性環(huán)氧化物水解酶抑制劑TPPU能夠保護(hù)MCAO大鼠血腦屏障完整性,維持其屏障功能,減輕腦水腫,減小腦梗死體積,減輕神經(jīng)功能缺損癥狀;上述作用具有劑量依賴性。
[Abstract]:Aim: to establish a rat model of cerebral ischemia / reperfusion (I / R) injury with thread embolization, and to regulate the cytochrome P450 epioxygenase CYP-EETs signaling pathway. To study the effect of soluble surface oxide hydrolase inhibitor (sEHi) TPPU) on the damage of blood-brain barrier (BBB) induced by cerebral ischemia / reperfusion injury, and to explore a new target for the treatment of cerebral ischemia. Methods: MCAO cerebral ischemia / reperfusion model was established by thread occlusion in rats. Rats were randomly divided into 5 groups: sham group, MCAO group (, TPPU (1.0mg/kg) group (, TPPU (0.3mg/kg) group (, TPPU (0.1mg/kg) group; TTC method was used to measure the infarct volume of MCAO, Yi Wen's blue method was used to detect the permeability of blood-brain barrier, and immunofluorescence staining was used. Western blot and other methods were used to detect the changes of blood-brain barrier related proteins after intervention of MCAO and TPPU. Brain edema was evaluated by dry and wet weight method and neurological functional defect and recovery was evaluated by Garcia18 score. Results: the success rate of t-MCAO model was high and the mortality was low. Laser Doppler Flowmeter could dynamically reflect the changes of cerebral blood flow in the middle cerebral artery (MCA) region of MCA (middle cerebral artery. It was helpful to judge whether MCA was an effective obstruction. Laser speckle imaging can visualize the changes of blood flow in the area of interest, but the disadvantages are that the blood flow changes in the region of interest can not be continuously monitored without continuous monitoring. After MCAO, there is a significant difference between the two groups (P0.01). In MCAO rats, the blood-brain barrier permeability was increased, a large number of Evans blue leakage was observed in the infarcted brain tissue, and then vascular cerebral edema was found, and TTC staining reflected the infarct size directly, which was consistent with the severity of nerve function defect. Compared with MCAO group, soluble epoxide hydrolase inhibitor (TPPU) could decrease the volume of cerebral cortex infarction (P0.05), but had no obvious effect on striatum infarction (P0.05). TPPU can reduce the loss of Blood-brain Barrier tight junction (tight junction,TJ) protein after MCAO (P0.05), which can help to maintain the integrity of BBB and reduce the leakage of Evans blue to brain tissue (P0.001). Then the vascular brain edema was alleviated (P0.05). TPPU could improve the symptoms of neurological deficit after cerebral infarction in MCAO rats, which was significantly higher in each dose group than that in the control group (P0.01), and the above benefits increased with the increase of TPPU dose in a dose-dependent manner. Conclusion: soluble epoxide hydrolase inhibitor (TPPU) can protect the integrity of blood-brain barrier, maintain its barrier function, reduce cerebral edema, reduce cerebral infarction volume and alleviate neurological deficit symptoms in MCAO rats, and these effects are dose-dependent.
【學(xué)位授予單位】：湖北醫(yī)藥學(xué)院
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R743.3

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 趙志剛,龔凌志;血腦屏障及藥物通過(guò)血腦屏障方法研究進(jìn)展[J];中國(guó)藥學(xué)雜志;2000年04期

2 秦鑫,張英起,顏真;跨越血腦屏障[J];生命的化學(xué);2003年03期

3 胡俊;血腦屏障開放的檢測(cè)方法[J];重慶醫(yī)學(xué);2003年12期

4 李吉華,周益群,張東友;中醫(yī)藥對(duì)血腦屏障影響的研究進(jìn)展[J];中國(guó)中西醫(yī)結(jié)合影像學(xué)雜志;2004年02期

5 李建生,王明航;血腦屏障在中醫(yī)藥研究中的地位及意義[J];中國(guó)中西醫(yī)結(jié)合雜志;2004年03期

6 金人一;;使藥物越過(guò)血腦屏障[J];生物技術(shù)通報(bào);1989年11期

7 ;什么是“血腦屏障”[J];人人健康;1996年02期

8 徐偉,地石;血腦屏障[J];中國(guó)神經(jīng)科學(xué)雜志;2000年02期

9 郭志儒;抗壞血酸能促進(jìn)藥物通過(guò)血腦屏障[J];中國(guó)獸醫(yī)學(xué)報(bào);2002年02期

10 王勁;顱內(nèi)原發(fā)性淋巴瘤開放血腦屏障后化療[J];中華神經(jīng)外科雜志;2004年02期

相關(guān)會(huì)議論文 前10條

1 肖瑩瑩;張藝;張靜;鄧，

本文編號(hào)：2303049