本文選題：重組人血管內(nèi)皮抑制素 + 平陽霉素�。� 參考：《廣西醫(yī)科大學(xué)》2011年碩士論文

【摘要】：目的：比較重組人血管內(nèi)皮抑制素、平陽霉素單獨使用及聯(lián)合應(yīng)用對人臍靜脈內(nèi)皮細(xì)胞系HUVEC細(xì)胞的作用效果,探討相關(guān)作用機制,以揭示內(nèi)皮抑素應(yīng)用于增生期血管瘤治療的可行性。 方法：傳代培養(yǎng)HUVEC細(xì)胞,取3-5代生長良好的細(xì)胞,將不同藥物濃度內(nèi)皮抑素、平陽霉素分別作用于HUVEC細(xì)胞,以四唑鹽比色法(MTT法)比較兩藥作用24h、48h后對HUVEC細(xì)胞的細(xì)胞抑制率,蘇木精-伊紅(HE)染色光鏡下觀察藥物作用前后細(xì)胞形態(tài)變化,用免疫組化法檢測藥物作用前后HUVEC細(xì)胞內(nèi)血管內(nèi)皮生長因子(VEGF)陽性表達(dá)的差異,流式細(xì)胞儀分析藥物作用后HUVEC細(xì)胞的細(xì)胞凋亡情況。 結(jié)果：內(nèi)皮抑素、平陽霉素分別作用HUVEC細(xì)胞后的吸光度值(OD值)均隨藥物濃度增大而減小,兩藥對HUVEC細(xì)胞的抑制作用表現(xiàn)出一定的濃度依賴性和時間依賴性。VEGF免疫組化染色顯示兩種藥物分別作用后HUVEC細(xì)胞的細(xì)胞胞漿灰度值均增大。光鏡下可見：內(nèi)皮抑素作用后HUVEC細(xì)胞形態(tài)明顯改變,蓋玻片上可看到大量凋亡細(xì)胞,并隨藥物濃度增大而增多,平陽霉素作用后大量細(xì)胞懸浮,貼壁細(xì)胞數(shù)量明顯減少,藥物濃度越大而貼壁細(xì)胞越少。流式細(xì)胞儀測得兩種藥物單獨使用均會引起HUVEC細(xì)胞凋亡,且均表現(xiàn)為濃度越大凋亡細(xì)胞數(shù)越多；兩種藥物聯(lián)合使用表現(xiàn)出對HUVEC細(xì)胞的協(xié)同抑制效應(yīng)。 結(jié)論：內(nèi)皮抑素、平陽霉素對HUVEC細(xì)胞均有明顯抑制作用,均能引起細(xì)胞凋亡,均阻止VEGF進入HUVEC細(xì)胞內(nèi)從而抑制細(xì)胞的增殖,說明兩藥對血管內(nèi)皮細(xì)胞的抑制作用的相關(guān)機制存在某種共性；較高濃度的內(nèi)皮抑素仍以細(xì)胞凋亡為主,而平陽霉素濃度大于一定范圍(20μg/ml)細(xì)胞抑制逐漸以壞死占主導(dǎo)；平陽霉素對血管內(nèi)皮細(xì)胞貼壁的影響較內(nèi)皮抑素明顯,揭示兩種藥物對血管內(nèi)皮細(xì)胞的抑制作用存在差異；內(nèi)皮抑素與平陽霉素的聯(lián)合使用對內(nèi)皮細(xì)胞的抑制有協(xié)同作用,為兩種藥物聯(lián)合使用治療增殖期血管瘤提供基礎(chǔ)理論依據(jù)。
[Abstract]:Aim: to compare the effects of recombinant human vascular endotheliostatin and pingyangmycin on human umbilical vein endothelial cell line HUVEC. To explore the feasibility of endostatin in the treatment of hyperplastic hemangioma. Methods: HUVEC cells were cultured and cultured in 3-5 passages. Different concentrations of endostatin and pingyangmycin were used to treat HUVEC cells respectively. The inhibitory rates of the two drugs on HUVEC cells after 24 h or 48 h treatment were compared by MTT assay. The morphological changes of HUVEC cells were observed under light microscope with hematoxylin and eosin (HEH) staining. The positive expression of vascular endothelial growth factor (VEGF) in HUVEC cells was detected by immunohistochemical method. Apoptosis of HUVEC cells was analyzed by flow cytometry. Results: the absorbance value and OD value of HUVEC cells treated with endostatin and pingyangmycin decreased with the increase of drug concentration. The inhibitory effects of the two drugs on HUVEC cells were concentration-dependent and time-dependent. VEGF immunohistochemical staining showed that the cytoplasmic gray values of HUVEC cells increased after the two drugs were treated separately. Under the light microscope, the morphology of HUVEC cells was obviously changed after the treatment of endostatin, a large number of apoptotic cells could be seen on the cover glass and increased with the increase of drug concentration. After the treatment of pingyangmycin, a large number of cells suspended and the number of adherent cells decreased obviously. The higher the drug concentration, the fewer adherent cells. The results of flow cytometry showed that both drugs alone could induce apoptosis of HUVEC cells, and the higher the concentration of the two drugs, the more the number of apoptotic cells, and the synergistic inhibitory effect of the two drugs on HUVEC cells. Conclusion: both endostatin and pingyangmycin can inhibit the proliferation of HUVEC cells, induce apoptosis, prevent VEGF from entering into HUVEC cells and inhibit the proliferation of HUVEC cells. The results showed that there was some common mechanism in the inhibition of vascular endothelial cells by the two drugs, the higher concentration of endostatin was still mainly apoptosis, while the inhibition of pingyangmycin was more than 20 渭 g / ml. The effect of pingyangmycin on the adhesion of vascular endothelial cells was more obvious than that on endostatin, which revealed that the inhibitory effects of two drugs on vascular endothelial cells were different, and the combination of endostatin and pingyangmycin had synergistic effects on the inhibition of endothelial cells. To provide basic theoretical basis for the combined use of two drugs in the treatment of proliferative hemangioma.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R329.3

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