血管內(nèi)皮抑制因子IFN-α和IFN-γ對淋巴管內(nèi)皮細(xì)胞生成的影響
發(fā)布時間：2018-08-07 14:21
【摘要】：目的 探討IFN-α(α-干擾素)和IFN-γ(γ-干擾素)對淋巴管內(nèi)皮細(xì)胞增殖和游走的影響及作用機(jī)制。旨在找到安全、有效、實用的淋巴管生成抑制劑。 材料和方法 本實驗所用淋巴管內(nèi)皮細(xì)胞來自豬的胸導(dǎo)管。1、淋巴管內(nèi)皮細(xì)胞的鑒定:應(yīng)用LYVE-1(淋巴管內(nèi)皮透明質(zhì)酸受體1)和VEGFR-3(血管內(nèi)皮生長因子受體-3)特異性抗體進(jìn)行鑒定。2、光鏡和電鏡觀察淋巴管內(nèi)皮細(xì)胞的形態(tài)和超微結(jié)構(gòu)。3、抑制實驗:本實驗分為兩大組:IFN-α組和IFN-γ組。每大組分別采用劃線法和MTT法觀察兩種因子對淋巴管內(nèi)皮細(xì)胞增殖和游走的影響。(1)劃線法:IFN-α組和IFN-γ組分別包括各自的對照組和3種濃度的實驗組。(2)MTT法:IFN-α組和IFN-γ組包括對照組和6種濃度的實驗組。4、細(xì)胞凋亡實驗:應(yīng)用Caspase染色法和Hoechst染色法進(jìn)行細(xì)胞凋亡實驗。5、干擾素活體實驗:在活體兔的兩個后肢上切斷淋巴管并分別注射IFN-α和生理鹽水,以觀察淋巴管的再生情況。 結(jié)果 1、淋巴管內(nèi)皮細(xì)胞的鑒定:經(jīng)LYVE-1鑒定和VEGFR-3鑒定,為典型淋巴管內(nèi)皮細(xì)胞。2、光鏡和電鏡觀察:光鏡下淋巴管內(nèi)皮細(xì)胞呈特征性“鵝卵石狀”或“鋪路石狀”鑲嵌排列;電鏡下觀察到淋巴管內(nèi)皮細(xì)胞的超微結(jié)構(gòu),細(xì)胞間有類似橋粒樣結(jié)構(gòu)。3、劃線法:與對照組相比,當(dāng)IFN-α和IFN-γ的濃度達(dá)到1000ng/ml時,兩者對淋巴管內(nèi)皮細(xì)胞的增殖和游走都有明顯的抑制作用(P0.01)。4、MTT法:當(dāng)IFN-α和IFN-γ的濃度達(dá)到2000ng/ml時,兩者對內(nèi)皮細(xì)胞的增殖具有顯著的抑制作用(P0.01)。5、凋亡實驗:(1)Caspase染色法證實,經(jīng)IFN-α和IFN-γ處理后的淋巴管內(nèi)皮細(xì)胞,陽性表達(dá)為胞漿呈黃色或棕黃色染色。(2)Hoechst染色證實,經(jīng)IFN-α和IFN-γ處理后的淋巴管內(nèi)皮細(xì)胞,可觀察到其核周圍有凋亡小體。6、干擾素活體實驗:手術(shù)切斷兔后肢淋巴管后16天,生理鹽水側(cè)淋巴管已愈合,無染料滲漏,IFN-α側(cè)仍有大量染料滲漏。 結(jié)論 1、IFN-α和IFN-γ對淋巴管內(nèi)皮細(xì)胞的增殖和游走具有明顯的抑制作用。2、IFN-α和IFN-γ具有促進(jìn)淋巴管內(nèi)皮細(xì)胞凋亡的作用。
[Abstract]:Objective to investigate the effect and mechanism of IFN- 偽 and IFN- 緯 on proliferation and migration of lymphatic endothelial cells. Aim to find safe, effective, and practical lymphangiogenesis inhibitors. Materials and methods Lymphatic endothelial cells from porcine thoracic ducts were identified by LYVE-1 (lymphatic endothelial hyaluronic acid receptor-1) and VEGFR-3 (vascular endothelial growth factor receptor -3) specific antibodies. The morphology and ultrastructure of lymphatic endothelial cells were observed by light microscope and electron microscope, and the inhibition experiment was carried out. The experiment was divided into two groups: group 1: IFN- 偽 and group IFN- 緯. The effects of two kinds of factors on the proliferation and migration of lymphatic endothelial cells were observed by underlined method and MTT method respectively. (1) the two groups included their respective control groups and three experimental groups with different concentrations. (2) the MTT method: IFN- 偽 group and IFN- 緯 group were divided into two groups: (1) the control group and the IFN- 緯 group, respectively. (2) the MTT method: IFN- 偽 group and IFN- 緯 group, respectively. The apoptosis test was carried out by Caspase staining and Hoechst staining. Interferon assay was performed in vivo. The lymphatic vessels were severed on the hind limbs of living rabbits and IFN- 偽 and normal saline were injected respectively. To observe the regeneration of lymphatic vessels. Results 1. The identification of lymphatic endothelial cells: by LYVE-1 and VEGFR-3 identification, they were typical lymphatic endothelial cells. Light and electron microscope observation showed that the lymphatic endothelial cells were cobblestone or paving stone mosaic. The ultrastructure of lymphatic endothelial cells was observed under electron microscope, and the desmosome like structure was observed between the cells. The results showed that compared with the control group, the concentration of IFN- 偽 and IFN- 緯 reached the level of 1000ng/ml, and the expression of IFN- 偽 and IFN- 緯 was similar to that of the control group. When the concentration of IFN- 偽 and IFN- 緯 reached 2000ng/ml, both of them could significantly inhibit the proliferation of endothelial cells (P0.01) .5. the apoptosis experiment: (1) Caspase staining showed that, when the concentration of IFN- 偽 and IFN- 緯 reached the level of 2000ng/ml, both of them could inhibit the proliferation of endothelial cells significantly (P0.01) .5. the apoptosis experiment: (1) Caspase staining confirmed that the proliferation of endothelial cells was inhibited by IFN- 偽 and IFN- 緯 (P0.01). The lymphatic endothelial cells treated with IFN- 偽 and IFN- 緯 were positive for yellow or brown cytoplasm staining. (2) Hoechst staining confirmed that the lymphatic endothelial cells were treated with IFN- 偽 and IFN- 緯. It was observed that there were apoptotic corpuscles. 6 around the nucleus. Interferon in vivo experiment: 16 days after the operation, the lymphatic vessels of the hind limbs of rabbits were healed, and a large amount of dye leakage was still found in the IFN- 偽 side of IFN- 偽 without dye leakage. Conclusion (1) IFN- 偽 and IFN- 緯 can significantly inhibit the proliferation and migration of lymphatic endothelial cells. (2) IFN- 偽 and IFN- 緯 can promote the apoptosis of lymphatic endothelial cells.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329
本文編號：2170303
[Abstract]:Objective to investigate the effect and mechanism of IFN- 偽 and IFN- 緯 on proliferation and migration of lymphatic endothelial cells. Aim to find safe, effective, and practical lymphangiogenesis inhibitors. Materials and methods Lymphatic endothelial cells from porcine thoracic ducts were identified by LYVE-1 (lymphatic endothelial hyaluronic acid receptor-1) and VEGFR-3 (vascular endothelial growth factor receptor -3) specific antibodies. The morphology and ultrastructure of lymphatic endothelial cells were observed by light microscope and electron microscope, and the inhibition experiment was carried out. The experiment was divided into two groups: group 1: IFN- 偽 and group IFN- 緯. The effects of two kinds of factors on the proliferation and migration of lymphatic endothelial cells were observed by underlined method and MTT method respectively. (1) the two groups included their respective control groups and three experimental groups with different concentrations. (2) the MTT method: IFN- 偽 group and IFN- 緯 group were divided into two groups: (1) the control group and the IFN- 緯 group, respectively. (2) the MTT method: IFN- 偽 group and IFN- 緯 group, respectively. The apoptosis test was carried out by Caspase staining and Hoechst staining. Interferon assay was performed in vivo. The lymphatic vessels were severed on the hind limbs of living rabbits and IFN- 偽 and normal saline were injected respectively. To observe the regeneration of lymphatic vessels. Results 1. The identification of lymphatic endothelial cells: by LYVE-1 and VEGFR-3 identification, they were typical lymphatic endothelial cells. Light and electron microscope observation showed that the lymphatic endothelial cells were cobblestone or paving stone mosaic. The ultrastructure of lymphatic endothelial cells was observed under electron microscope, and the desmosome like structure was observed between the cells. The results showed that compared with the control group, the concentration of IFN- 偽 and IFN- 緯 reached the level of 1000ng/ml, and the expression of IFN- 偽 and IFN- 緯 was similar to that of the control group. When the concentration of IFN- 偽 and IFN- 緯 reached 2000ng/ml, both of them could significantly inhibit the proliferation of endothelial cells (P0.01) .5. the apoptosis experiment: (1) Caspase staining showed that, when the concentration of IFN- 偽 and IFN- 緯 reached the level of 2000ng/ml, both of them could inhibit the proliferation of endothelial cells significantly (P0.01) .5. the apoptosis experiment: (1) Caspase staining confirmed that the proliferation of endothelial cells was inhibited by IFN- 偽 and IFN- 緯 (P0.01). The lymphatic endothelial cells treated with IFN- 偽 and IFN- 緯 were positive for yellow or brown cytoplasm staining. (2) Hoechst staining confirmed that the lymphatic endothelial cells were treated with IFN- 偽 and IFN- 緯. It was observed that there were apoptotic corpuscles. 6 around the nucleus. Interferon in vivo experiment: 16 days after the operation, the lymphatic vessels of the hind limbs of rabbits were healed, and a large amount of dye leakage was still found in the IFN- 偽 side of IFN- 偽 without dye leakage. Conclusion (1) IFN- 偽 and IFN- 緯 can significantly inhibit the proliferation and migration of lymphatic endothelial cells. (2) IFN- 偽 and IFN- 緯 can promote the apoptosis of lymphatic endothelial cells.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R329
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 曹琦珍,林志彬;IFNγ在抗腫瘤新生血管形成中潛在的作用及機(jī)制研究[J];中國臨床藥理學(xué)與治療學(xué);2004年08期
2 郭向陽,康驊;淋巴管內(nèi)皮特異性標(biāo)記物及其在腫瘤淋巴管生成研究中的應(yīng)用[J];腫瘤防治研究;2005年06期
相關(guān)碩士學(xué)位論文 前1條
1 遲曉艷;血管內(nèi)皮抑制因子angiostatin和thalidomide對淋巴管內(nèi)皮細(xì)胞生成的影響[D];青島大學(xué);2005年
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