本文關(guān)鍵詞： 動(dòng)脈粥樣硬化 血管細(xì)胞黏附因子1 纖溶酶原激活劑抑制劑1 通心絡(luò)　出處：《吉林大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：背景與目的： AS已成為威脅人類健康,引起死亡的主要疾病。可導(dǎo)致全身多個(gè)靶器官損害,嚴(yán)重影響人民的生活質(zhì)量。為從根本解決AS為人民帶來的困擾,大量基礎(chǔ)實(shí)驗(yàn)及臨床觀察致力于對(duì)AS發(fā)病機(jī)制及治療方法的研究。本研究通過高脂飲食建立家兔AS模型,了解PAI-1和VCAM-1在AS家兔血清和病變局部的表達(dá),觀察通心絡(luò)對(duì)PAI-1和VCAM-1的影響及其抗AS的作用機(jī)制。 資料與方法： 一、建立家兔AS模型,采用酶聯(lián)免疫夾心法、免疫組化方法檢測(cè)VCAM-1和PAI-1在正常飲食組和高脂飲食組家兔血清中及動(dòng)脈血管壁的表達(dá)。 (一)動(dòng)物模型的建立 1、實(shí)驗(yàn)動(dòng)物和分組 雄性大耳白兔24只。隨機(jī)分為常規(guī)飲食組、高脂飲食組、通心絡(luò)組,每組8只,飼養(yǎng)14周。 2、病理標(biāo)本的制作 14周末處死動(dòng)物,常規(guī)病理切片,H-E染色。 3、高脂血癥的確立 正常飲食組和高脂飲食組實(shí)驗(yàn)前(0周)血清中TC、TG、LDL無顯著性差異(P0.01)；正常飲食組14周TC、TG、LDL與0周比較無顯著性差異(P0.01)；高脂飲食組14周TC、TG、LDL明顯增高,與0周比較有顯著性差異(P0.01)。 (二)酶聯(lián)免疫夾心法檢測(cè)VCAM-1和PAI-1在正常飲食組和高脂飲食組血清中的表達(dá) 1、VCAM-1在正常飲食組和高脂飲食組血清中的表達(dá) 正常飲食組的血清中VCAM-1水平在0周、14周無顯著差別(P0.01)；高脂飲食組血清中VCAM-1水平14周較0周顯著增高(P0.01)。 2. PAI-1在正常飲食組和高脂飲食組血清中的表達(dá) 正常飲食組血清中PAI-1水平在0周、14周無顯著差別(P0.01);高脂飲食組血清中PAI-1的水平14周較0周顯著增高(P0.01)。 (三)免疫組化檢測(cè)VCAM-1和PAI-1在正常飲食組和高脂飲食組主動(dòng)脈壁的表達(dá) 1、VCAM-1在正常飲食組和高脂飲食組主動(dòng)脈壁的陽性表達(dá) 正常飲食組主動(dòng)脈內(nèi)膜和中膜僅見散在棕黃色顆粒狀陽性染色物質(zhì),陽性染色百分比為2.92±0.31%。高脂飲食組增厚的動(dòng)脈內(nèi)膜及中膜內(nèi)可見大量的棕黃色顆粒狀陽性染色物質(zhì),陽性染色百分比為18.38±2.55%,明顯高于正常飲食組(P0.01)。 2、PAI-1在正常飲食組和高脂飲食組主動(dòng)脈壁的陽性表達(dá) 正常飲食組主動(dòng)脈內(nèi)膜和中膜僅見散在棕黃色顆粒狀陽性染色物質(zhì),陽性染色百分比為3.24±0.21%。高脂飲食組增厚的動(dòng)脈內(nèi)膜及中膜內(nèi)可見大量的棕黃色顆粒狀陽性染色物質(zhì),尤以內(nèi)膜明顯,陽性染色百分比為15.46±2.23%,明顯高于正常飲食組(P0.01)。 二、通心絡(luò)的調(diào)脂作用 實(shí)驗(yàn)前三組動(dòng)物血清TC、TG、LDL無差異(P0.01),14周時(shí)正常飲食組TC、TG、LDL與0周比較無顯著性差異(P0.01)；14周時(shí)高脂飲食組和通心絡(luò)組TC、TG、LDL明顯增高,明顯高于正常飲食組(P0.01)；高脂飲食組和通心絡(luò)組比較,14周時(shí)通心絡(luò)組血清TC、TG、LDL較高脂飲食組明顯下降(P0.01)。 三、通心絡(luò)對(duì)VCAM-1和PAI-1表達(dá)的影響 (一)通心絡(luò)對(duì)血清VCAM-1和PAI-1表達(dá)的影響 1、通心絡(luò)對(duì)血清VCAM-1表達(dá)的影響 三組動(dòng)物血清中VCAM-1水平在0周無顯著差別(P0.01)；飼養(yǎng)14周時(shí)高脂飲食組和通心絡(luò)組血清中VCAM-1水平較0周顯著增高(P0.01),并高于正常飲食組(P0.01)；14周時(shí)通心絡(luò)組血清中VCAM-1較高脂飲食組明顯下降(P0.01)。 2、通心絡(luò)對(duì)血清PAI-1表達(dá)的影響 三組動(dòng)物血清中PAI-1水平在0周無顯著差別(P0.01)；飼養(yǎng)14周時(shí)高脂飲食組和通心絡(luò)組血清中PAI-1水平較0周增高(P0.01),并高于正常飲食組(P0.01)；14周時(shí)通心絡(luò)組血清中PAI-1較高脂飲食組明顯下降(P0.01)。 (二)通心絡(luò)對(duì)主動(dòng)脈壁VCAM-1和PAI-1陽性染色百分比的影響 1、通心絡(luò)對(duì)主動(dòng)脈壁VCAM-1陽性染色百分比的影響 高脂飲食組和通心絡(luò)組增厚的動(dòng)脈內(nèi)膜及中膜內(nèi)可見較多的棕黃色顆粒狀陽性染色物質(zhì),表達(dá)量分別為18.38±2.55%和13.18±1.25%。通心絡(luò)組的陽性表達(dá)百分比較高脂飲食組明顯降低(P0.01)。 2、通心絡(luò)對(duì)主動(dòng)脈壁PAI-1陽性染色百分比的影響 高脂飲食組和通心絡(luò)組增厚的動(dòng)脈內(nèi)膜及中膜內(nèi)可見較多的棕黃色顆粒狀陽性染色物質(zhì),表達(dá)量分別為15.46±2.23%和9.45±1.09%。通心絡(luò)組的陽性表達(dá)百分比較高脂飲食組明顯降低(P0.01)。 以上實(shí)驗(yàn)結(jié)果以平均數(shù)±標(biāo)準(zhǔn)差(X±S)表示,自身及組間對(duì)照采用t檢驗(yàn),P0.05有顯著性差異。結(jié)果： 1.高脂飲食組和通心絡(luò)組14周后血清TC、TG、LDL較實(shí)驗(yàn)前明顯升高(P0.01),并且高于對(duì)照組(P0.01),通心絡(luò)組血清TC、TG、LDL較高脂飲食組明顯下降(P0.01)。 2.高脂飲食組和通心絡(luò)組14周后血清PAI-1和VCAM-1的水平較0周顯著增高(P0.01),并且高于對(duì)照組(P0.01),通心絡(luò)組PAI-1和VCAM-1的水平明顯低于高脂飲食組(P0.01)。 3.免疫組化分析顯示：高脂飲食組增厚的動(dòng)脈內(nèi)膜及中膜內(nèi)可見大量的棕黃色顆粒狀陽性染色物質(zhì),部分融合成片,尤以內(nèi)膜明顯,通心絡(luò)組陽性染色物質(zhì)明顯低于高脂飲食組,三組組間比較有顯著性差異(P0.01)。 結(jié)論： 1、AS家兔血清中VCAM-1和PAI-1的表達(dá)增高；家兔主動(dòng)脈粥樣硬化病變局部VCAM-1和PAI-1陽性表達(dá)百分比增高。VCAM-1和PAI-1的過度表達(dá)可能是AS發(fā)生機(jī)制之一。 2、通心絡(luò)能夠減少AS家兔VCAM-1和PAI-1在血清中及主動(dòng)脈粥樣硬化病變局部陽性表達(dá)百分比。抑制VCAM-1和PAI-1的表達(dá)可能是通心絡(luò)抗AS機(jī)制之一。 3、通心絡(luò)不但具有調(diào)脂的作用,并且具有減輕和抑制AS的作用。 4、檢測(cè)血清中VCAM-1和PAI-1,對(duì)于早期診斷AS性疾病以及監(jiān)測(cè)AS的發(fā)生、發(fā)展及轉(zhuǎn)歸有著重要的意義。
[Abstract]:Background and purpose:
AS has become a threat to human health, mainly caused by the disease can lead to death. More than one target organ damage, seriously affecting people's quality of life. In order to fundamentally solve the AS problems for people, a large number of basic experiments and clinical observation of AS is devoted to the study of pathogenesis and therapy. The study of rabbit AS model the high fat diet, understand PAI-1 and VCAM-1 expression in serum and AS local lesions in rabbits, observe the mechanism effect of Tongxinluo on PAI-1 and VCAM-1 and anti AS.
Information and methods:
First, the rabbit AS model was established. The expression of VCAM-1 and PAI-1 in serum and arterial wall of normal diet group and high-fat diet group were detected by enzyme-linked immunosorbent assay and immunohistochemistry.
(I) establishment of animal model
1, experimental animals and groups
24 male large white rabbits were randomly divided into regular diet group, high fat diet group, Tongxinluo group and 8 rats in each group for 14 weeks.
2, the preparation of pathological specimen
The animals were killed at the end of the 14 week, routine pathological section, H-E staining.
3, the establishment of hyperlipidemia
There was no significant difference in serum TC, TG and LDL between the normal diet group and the high-fat diet group before the experiment (0 weeks) (P0.01). There was no significant difference in TC, TG, LDL between the normal diet group and the 0 week group (P0.01). The 14 week TC, TG and LDL increased significantly in the high-fat diet group, which was significantly different from that in the 0 week (TC).
(two) the expression of VCAM-1 and PAI-1 in the serum of the normal diet group and the high fat diet group by enzyme linked immunosorbent assay
1, the expression of VCAM-1 in the serum of the normal diet group and the high fat diet group
The level of VCAM-1 in the serum of the normal diet group was 0 weeks, and there was no significant difference in the 14 weeks (P0.01), and the level of VCAM-1 in the serum of high fat diet group was significantly higher than that in the 0 week (P0.01).
2. PAI-1 expression in the serum of the normal diet group and the high fat diet group
There was no significant difference in the level of PAI-1 in the serum of the normal diet group for 0 weeks and 14 weeks (P0.01), and the level of PAI-1 in the serum of the high fat diet group was significantly higher than that in the 0 week (P0.01).
(three) immunohistochemical detection of the expression of VCAM-1 and PAI-1 in the aorta wall of the normal diet group and the high fat diet group
1, the positive expression of VCAM-1 in the aorta wall of the normal diet group and the high fat diet group
Normal diet group aortic intima and media were scattered in the yellow brown granular staining material, positive staining percentage was 2.92 + 0.31%. arterial intimal thickening of the high fat diet group and in the film can be seen in the large brown granular staining material, positive staining percentage was 18.38 + 2.55%, significantly higher than the normal diet group (P0.01).
2, the positive expression of PAI-1 in the aorta wall of the normal diet group and the high fat diet group
Normal diet group aortic intima and media were scattered in the yellow brown granular staining material, positive staining percentage was 3.24 + 0.21%. arterial intimal thickening of the high fat diet group and in the film can be seen in the large brown granular staining material, especially in endometrial. Positive staining percentage was 15.46 + 2.23%, significantly higher than the normal diet group (P0.01).
Two, the lipid-regulating effect of Tongxinluo
Before the experiment three group animal serum TC, TG, LDL no difference (P0.01), TC, normal diet group at 14 weeks compared with 0 weeks TG, LDL no significant difference (P0.01); 14 weeks of high fat diet group and Tongxinluo group TC, TG, LDL increased significantly, significantly higher than the normal diet group (P0.01); high fat diet group and Tongxinluo group, 14 weeks Shitong Tongxinluo group serum TC, TG, LDL in high fat diet group decreased significantly (P0.01).
Three, the effect of Tongxinluo on the expression of VCAM-1 and PAI-1
(1) the effect of Tongxinluo on the expression of serum VCAM-1 and PAI-1
1, the effect of Tongxinluo on the expression of serum VCAM-1
The VCAM-1 levels of the three groups in animal serum had no significant difference in the 0 week (P0.01); 14 weeks of feeding, high fat diet group and Tongxinluo group serum VCAM-1 levels were significantly increased in 0 weeks (P0.01), and higher than the normal diet group (P0.01); VCAM-1 high fat diet for 14 weeks Shitong Tongxinluo group decreased (P0.01).
2, the effect of Tongxinluo on the expression of serum PAI-1
The PAI-1 levels of the three groups in animal serum had no significant difference in the 0 week (P0.01); 14 weeks of feeding, high fat diet group and Tongxinluo group in the serum PAI-1 level was 0 Zhou Zenggao (P0.01), and higher than the normal diet group (P0.01); PAI-1 high fat diet for 14 weeks Shitong Tongxinluo group was significantly decreased (P0.01).
(two) the effect of Tongxinluo on the percentage of VCAM-1 and PAI-1 positive staining in the aortic wall
1, the effect of Tongxinluo on the percentage of VCAM-1 positive staining in the aortic wall
In the high fat diet group and Tongxinluo group, there were more brown and granular positive staining substances in the intima and middle membrane. The expression levels were 18.38 + 2.55% and 13.18 + 1.25%. respectively. The percentage of positive expression in Tongxinluo group was significantly lower than that in the high-fat diet group (P0.01).
2, the effect of Tongxinluo on the percentage of PAI-1 positive staining in the aortic wall
In the high fat diet group and Tongxinluo group, there were more brown and granular positive staining substances in the intima and middle membrane. The expression levels were 15.46 + 2.23% and 9.45 + 1.09%. respectively. The percentage of positive expression in Tongxinluo group was significantly lower than that in the high-fat diet group (P0.01).
The above experimental results were expressed with mean standard deviation (X + S). T test was used in self and group control, and there were significant differences in P0.05.
1., after 14 weeks of high fat diet group and Tongxinluo group, serum TC, TG and LDL increased significantly (P0.01), which was higher than that of the control group (P0.01). The serum TC, TG and LDL of the Tongxinluo group were significantly lower than those of the high-fat diet group (P0.01).
2., after 14 weeks of high fat diet group and Tongxinluo group, serum PAI-1 and VCAM-1 levels increased significantly compared with 0 weeks (P0.01), and higher than those in control group (P0.01). The levels of PAI-1 and VCAM-1 in Tongxinluo group were significantly lower than those in high-fat diet group (P0.01).
3. immunohistochemical analysis showed that intimal thickening in cholesterol diet group and in the film can be seen in the large brown granular staining material, partial integration into the film, especially in endometrial. Tongxinluo group positive substance was significantly lower than that in high fat diet group, there were significant differences between the three groups (P0.01).
Conclusion:
1, the expression of VCAM-1 and PAI-1 increased in serum of AS rabbits. The percentage of positive expression of VCAM-1 and PAI-1 in aorta atherosclerotic lesion of rabbits increased, and the over expression of.VCAM-1 and PAI-1 might be one of the mechanisms of AS.
2, Tongxinluo can reduce the percentage of positive expression of VCAM-1 and PAI-1 in serum and atherosclerotic lesion in AS rabbits. Inhibition of VCAM-1 and PAI-1 expression may be one of the mechanisms of Tongxinluo AS.
3, Tongxinluo not only has the effect of lipid-regulating, but also has the effect of reducing and inhibiting AS.
4, the detection of VCAM-1 and PAI-1 in serum is of great significance for the early diagnosis of AS disease and the monitoring of the occurrence, development and prognosis of AS.

【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R-332

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