【摘要】：目的： 蛛網(wǎng)膜下腔出血(subarachnoid hemorrhage, SAH)是由于各種原因造成顱內(nèi)出血，血液流入蛛網(wǎng)膜下腔形成的。按其發(fā)生機制不同可分為：外傷性SAH、自發(fā)性SAH、繼發(fā)性SAH。外傷性SAH是顱腦外傷導(dǎo)致的；自發(fā)性SAH是指無明顯誘因SAH，主要顱內(nèi)動靜脈畸形或動脈瘤破裂出血造成；繼發(fā)性SAH是由于腦室內(nèi)或腦實質(zhì)內(nèi)出血彌散到蛛網(wǎng)膜下腔形成的。SAH后急性期神經(jīng)系統(tǒng)并發(fā)癥主要有腦血管痙攣造成的缺血性腦損傷、出血部位發(fā)生再出血、急性腦積水等。慢性腦積水是其遠(yuǎn)期主要并發(fā)癥之一，嚴(yán)重影響患者的預(yù)后、生活質(zhì)量。SAH作為神經(jīng)內(nèi)外科常見危重急癥，目前對急性期并發(fā)癥研究較多，對腦積水的研究相對較少，對腦積水發(fā)生的具體機制尚未明確。多數(shù)學(xué)者認(rèn)為：SAH后急性腦積水的發(fā)生是由于血凝塊堵塞腦脊液循環(huán)通路造成的梗阻性腦積水，起病比較急,主要表現(xiàn)為神情淡漠、反應(yīng)遲鈍、原有癥狀加重、重癥病人出現(xiàn)昏迷、意識障礙。SAH后慢性腦積水的發(fā)生主要原因在于出血造成的應(yīng)激、炎癥反應(yīng)刺激蛛網(wǎng)膜、軟腦膜、蛛網(wǎng)膜小梁等柔腦膜結(jié)構(gòu)發(fā)生纖維化病變，使局部結(jié)構(gòu)發(fā)生纖維化粘連，造成腦脊液的吸收減少，導(dǎo)致交通性腦積水；堵塞腦脊液循環(huán)通路造成腦脊液循環(huán)受阻，形成梗阻性腦積水。吡非尼酮(Pirfenidone, PFD)是一種小分子化合物，化學(xué)式：C12Hl1NO，PFD分子能夠通過細(xì)胞膜，無需受體�？诜o藥后經(jīng)胃腸道吸收，迅速在體內(nèi)各組織中廣泛分布，可透過血腦屏障。大量實驗證實PFD在抗纖維化治療方面的療效顯著且不良反應(yīng)少。主要應(yīng)用于特異性肺纖維化、腎臟、肝臟、心肌組織、多發(fā)硬化癥和神經(jīng)纖維瘤等抗纖維化疾病的治療研究。利用PFD預(yù)防及治療SAH后的柔腦膜纖維化、減低SAH后慢性腦積水的發(fā)生率的研究尚未見報道。本研究擬觀察PFD與大鼠SAH后腦脊液中強致纖維化因子之一轉(zhuǎn)化生長因子-β1（transforming growth factor beta1，TGF-β1）的關(guān)系，以及其對柔腦膜纖維化的影響。 方法: 采用清潔級SD大鼠80只，雌雄不限，體重250±10g，隨機分為正常對照組、假手術(shù)組和模型組；模型組又分為干預(yù)組和對照組。采用血管內(nèi)穿刺法建立SAH動物模型，干預(yù)組給予PFD100mg/kg灌胃給藥，對照組給予安慰劑灌胃，連續(xù)14天，分別在3、6、10、14、21天自枕大池采集腦脊液標(biāo)本，檢測各組腦脊液中TGF-β1的濃度；第21天將試驗動物灌注取腦，經(jīng)MASSON染色處理后，用病理圖文分析系統(tǒng)圖像處理軟件Image-Pro Plus6.0分析圖像，測量柔腦膜的厚度和灰度值，以100×厚度/灰度值作為膠原纖維含量的指標(biāo)，所得數(shù)據(jù)均采用SPSS13.0軟件進(jìn)行統(tǒng)計學(xué)分析。觀察其對柔腦膜纖維化的程度的影響。 結(jié)果: 1各組大鼠一般情況的觀察比較 空白組大鼠進(jìn)食好，精神好，被毛有光澤，活動靈活，未見明顯異常狀態(tài)，隨著飼養(yǎng)時間的延長，體型漸長，體重不斷增加。 假手術(shù)組大鼠麻醉清醒后表現(xiàn)為豎毛、精神萎靡、飲食攝水減少、活動減少，于術(shù)后第1日恢復(fù)正常，進(jìn)食增多，精神狀態(tài)佳，被毛光澤，活動靈活，無異常狀態(tài)出現(xiàn)，隨著飼養(yǎng)時間的延長，體型漸長，體重不斷增加。 模型組大鼠麻醉清醒后表現(xiàn)為豎毛、精神萎靡、嗜睡，定向力差、于術(shù)后1周內(nèi)均精神萎靡、飲食攝水減少，活動減少，于術(shù)后1周后漸恢復(fù)，精神較前好轉(zhuǎn)，飲食攝水增加，但與空白組及假手術(shù)組相比活動減少，反應(yīng)較遲鈍。隨著飼養(yǎng)時間延長，體型有所增長。 2模型制作觀察及神經(jīng)行為評分 模型制作24小時后，空白組和假手術(shù)組灌注取腦后，蛛網(wǎng)膜下腔未見血液成分，顱底未見血凝塊。模型組灌注取腦后大腦腹側(cè)面可見血凝塊存在，主要位于前顱窩及顱底動脈環(huán)周圍，包繞腦底部主要血管，，并且穿刺側(cè)出血量相對較多，顱底相對應(yīng)位置可見血凝塊。模型制作24小時候，參照Bederson方法進(jìn)行神經(jīng)行為功能缺損評分。將評分2分以上為納入模型成功標(biāo)準(zhǔn)。結(jié)果提示:空白組和假手術(shù)組沒有功能缺損，模型組神經(jīng)功能缺損評分平均為2.50±0.56分，與空白對照組進(jìn)行比較，評分有統(tǒng)計學(xué)意義。 3轉(zhuǎn)化因子-β1及柔腦膜纖維化影響 對照組大鼠腦脊液中TGF-β1的濃度出現(xiàn)2次升高，與空白組比較差異有顯著性（P0.05）。干預(yù)組腦脊液中TGF-β1的濃度也出現(xiàn)2次升高，但最高值明顯低于對照組（P0.05）�？瞻捉M、假手術(shù)組無明顯變化。MASSON染色顯示對照組與干預(yù)組較空白組、假手術(shù)組柔腦膜增厚且灰度值明顯減低（P0.05），其中干預(yù)組較對照組柔腦膜纖維化明顯減低（P0.05）�？瞻捉M與假手術(shù)組柔腦膜無明顯纖維化。 結(jié)論: 1SAH發(fā)生后腦脊液中TGF-β1濃度明顯升高，并呈雙時相性。 2SAH21天后可出現(xiàn)柔腦膜纖維化現(xiàn)象。 3SAH發(fā)生后應(yīng)用PFD干預(yù)可降低腦脊液中的TGF-β1的濃度，特別是能夠明顯減低第二時相的升高程度，能夠明顯減低柔腦膜的纖維化程度。
[Abstract]:Objective:
Subarachnoid hemorrhage (subarachnoid hemorrhage, SAH) is due to various causes causing intracranial hemorrhage and blood flow into the subarachnoid cavity. According to its mechanism, it can be divided into traumatic SAH, spontaneous SAH, and secondary SAH. traumatic SAH, which is caused by craniocerebral trauma; spontaneous SAH refers to no obvious inducement SAH and major intracranial movement. The secondary SAH is the ischemic brain injury caused by cerebral vasospasm, bleeding sites and acute hydrocephalus. Chronic hydrocephalus is the main complication of the acute phase of.SAH in the acute phase of the acute phase after the hemorrhage of the intraventricular or intraventricular hemorrhage to the subarachnoid cavity. One of the disease, which seriously affects the prognosis of the patients, the quality of life.SAH is a common critical emergency in the internal and external department. At present, there are many studies on the acute complications, the research on hydrocephalus is relatively few, the specific mechanism of hydrocephalus is not clear. Most scholars believe that the occurrence of acute hydrocephalus after SAH is due to the clogging of the brain ridge with blood clot. The obstructive hydrocephalus caused by the fluid circulation pathway is very urgent, mainly manifested in the indifference, the slow reaction, the aggravation of the original symptoms and the coma in the severe patients. The main cause of the chronic hydrocephalus after.SAH is the stress caused by bleeding, and the inflammatory reaction stimulates the arachnoid, the pia meninges, the arachnoid trabecula and so on. The occurrence of fibrosis, the local structure of fibrosis adhesion, resulting in the absorption of cerebrospinal fluid absorption, causing traffic hydrocephalus; blocking the cerebrospinal fluid circulation pathway caused by the cerebrospinal fluid circulation obstruction, the formation of obstructive hydrocephalus. Pirfenidone (PFD) is a small molecular compound, chemical formula: C12Hl1NO, PFD molecules can pass Cell membrane, without the need for receptor. After oral administration, it is absorbed by the gastrointestinal tract and is widely distributed in various tissues in the body and through the blood brain barrier. A large number of experiments have proved that PFD has significant effects on anti fibrosis and less adverse reactions. It is mainly used in specific pulmonary fibrosis, kidney, liver, myocardial tissue, multiple sclerosis and neurofibroma. Research on the treatment of anti fibrosis diseases. The study on the use of PFD to prevent and treat SAH after SAH has not been reported. This study is to observe the relationship between PFD and rat SAH, one of the strong fibrotic factors transforming growth factor - beta 1 (transforming growth factor beta1, TGF- beta 1). Line, and its effect on the fibrosis of the soft meninges.
Method:
80 SD rats were divided into the normal control group, the sham operation group and the model group, and the model group was divided into the intervention group and the control group. The model group was divided into the intervention group and the control group. The SAH animal model was established by intravascular puncture, the intervention group was given the medicine of PFD100mg/kg and the control group was given the placebo for 14 days, respectively, at 3,6,10,14, The concentration of TGF- beta 1 in cerebrospinal fluid was collected at 21 days from the big cistern, and the concentration of the cerebrospinal fluid in the cerebrospinal fluid was detected. After twenty-first days, the experimental animals were perfused to take the brain. After MASSON staining, the image processing software Image-Pro Plus6.0 was analyzed by the pathological graphic analysis system. The thickness and gray value of the soft meninges were measured, and the 100 x thickness / gray value was used as the collagen fiber. The data were analyzed by SPSS13.0 software. The effect of SPSS13.0 on the degree of leptomeningeal fibrosis was observed.
Result:
1 observation and comparison of the general situation of rats in each group
The rats in blank group had good eating, good spirit, glossy coat, flexible movement and no obvious abnormality. With the prolongation of feeding time, the body shape and weight increased gradually.
The rats in the sham operation group showed the erect hair after sober anesthesia, depressed spirit, reduced food intake, reduced activity, resumed normal in first days after the operation, increased eating and had better mental state, was flexible, and had no abnormal state. As the feeding time extended, the body type grew gradually, and the weight was constantly increasing.
The rats in the model group were shown to be erect, depressed, sleepy, and poor Orienteering after 1 weeks of anesthesia. They were depressed in the spirit, reduced food intake and decreased activity after 1 weeks of operation. The spirits were gradually recovered after 1 weeks, the spirit was better and the food intake was increased, but the activity was reduced and the reaction was slow compared with the blank group and the sham operation group. With the feeding time prolonged, There is a growth in shape.
2 model making observation and neurobehavioral score
After 24 hours of making the model, there was no blood component in the subarachnoid space and no blood clot in the subarachnoid space. The blood clot was found on the ventral side of the brain of the model group, mainly located around the anterior cranial fossa and the ring of the skull base artery. The blood vessels were wrapped around the base of the brain, and the amount of bleeding on the puncture side was relatively large, and the skull base was relatively large. The blood clot was seen in the relative position. When the model was made 24, the Bederson method was used to score the neurobehavioral function defect score. The score of the score was more than 2 points. The results showed that there was no functional defect in the blank group and the sham operation group, and the average score of the neural function defect was 2.50 + 0.56 points in the model group, compared with the blank control group. Compared, the score was statistically significant.
3 transformation factor - beta 1 and the influence of the soft meningeal fibrosis
The concentration of TGF- beta 1 in the cerebrospinal fluid of the control group increased 2 times, compared with the blank group (P0.05). The concentration of TGF- beta 1 in the cerebrospinal fluid of the intervention group was also increased 2 times, but the highest value was significantly lower than that of the control group (P0.05). There was no obvious change in the sham group and the.MASSON staining showed that the control group and the intervention group were more than the blank group and the false hand. In the operation group, the soft meninges were thickened and the gray value decreased significantly (P0.05), and the fibrosis in the intervention group was significantly lower than that in the control group (P0.05). There was no obvious fibrosis in the blank group and the sham group.
Conclusion:
The concentration of TGF- beta 1 in CSF increased significantly after 1SAH.
The phenomenon of soft meningeal fibrosis appears after 2SAH21 days.
The use of PFD intervention after 3SAH can reduce the concentration of TGF- beta 1 in cerebrospinal fluid, especially to reduce the level of second phase obviously, and reduce the degree of fibrosis of the meningomeninges.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743.35

【參考文獻(xiàn)】

相關(guān)期刊論文 前7條

1 高成;陳會榮;劉相軫;趙振環(huán);李敬文;;三種方法制作大鼠蛛網(wǎng)膜下腔出血模型[J];中國微侵襲神經(jīng)外科雜志;2008年09期

2 袁彬;李彤;;凝血酶對TGF-β_1和大鼠軟腦膜纖維化的相關(guān)性研究[J];中風(fēng)與神經(jīng)疾病雜志;2008年03期

3 劉亞武;祁吉;;動脈瘤性蛛網(wǎng)膜下腔出血的影像學(xué)診斷及治療[J];國際醫(yī)學(xué)放射學(xué)雜志;2008年01期

4 Franck Verrecchia;Alain Mauviel;;Transforming growth factor-β and fibrosis[J];World Journal of Gastroenterology;2007年22期

5 王國權(quán),吳浩,袁伯俊;吡非尼酮的動物急性毒性[J];藥學(xué)服務(wù)與研究;2005年01期

6 趙立衛(wèi),林成海,劉相軫;動脈瘤性蛛網(wǎng)膜下腔出血后腦積水[J];國外醫(yī)學(xué)(腦血管疾病分冊);2004年09期

7 馮文忠,王萬銘,王光海,李衛(wèi)華,唐榮華;實驗性SAH腦脊液TGF-β1檢測與慢性腦積水形成機制的研究[J];腦與神經(jīng)疾病雜志;2003年03期

本文編號：2168321