本文選題：青蒿素 + SCID小鼠�。� 參考：《昆明醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:首先觀察蒿甲醚在體外對U251人腦膠質(zhì)瘤細(xì)胞增殖的影響,隨后建立SCID小鼠人腦膠質(zhì)瘤原位模型,觀察蒿甲醚在體內(nèi)對膠質(zhì)瘤的抑制及其抗血管生成的作用,探討蒿甲醚抑制膠質(zhì)瘤增殖的機(jī)制,為膠質(zhì)瘤的臨床治療提供一種新的思路和方法。方法:1.體外細(xì)胞基中培養(yǎng)人腦U251膠質(zhì)瘤細(xì)胞,采用MTT(四甲基偶氮唑藍(lán))法測定蒿甲醚對U251膠質(zhì)瘤細(xì)胞的生長抑制作用,并計算半數(shù)抑制濃度(IC50);采用ELISA法檢測蒿甲醚抑制U251膠質(zhì)瘤細(xì)胞的VEGF表達(dá)。2.構(gòu)建SCID小鼠人腦膠質(zhì)瘤原位模型,采用立體定位儀定位麻醉狀態(tài)小鼠,剪開頭皮后,在距矢狀縫與冠狀縫交點處分別向右向后2mm定位注射點,鉆孔后,使用10ul微量注射器垂直于顱骨在小鼠腦部定位點(右側(cè)頂葉區(qū))深5mm處,注入10ulU251人腦膠質(zhì)瘤細(xì)胞,骨蠟填封骨孔,可吸收線縫合頭皮,觀察小鼠生活狀態(tài),在小鼠出現(xiàn)癲癇癥狀后,經(jīng)活體左心室灌注后取腦組織肉眼觀察腫瘤大體外觀及切面,并行HE染色確證腫瘤模型成功構(gòu)建。3.構(gòu)建24只SCID小鼠U251人腦膠質(zhì)瘤原位模型,隨機(jī)分為3組,分別為對照組(生理鹽水0.4ml/10g/d)、蒿甲醚組(蒿甲醚50.0mg/kg/d)、替莫唑胺組(20mg/kg/d)。替莫唑胺采用其粉劑制成混懸液灌胃,量為1.65mg/只/d。接種后標(biāo)記小鼠,每天定時定量給藥,并記錄生存時間。約接種20天后,當(dāng)SCID小鼠瀕臨死亡時行活體左心灌注,取出并固定全部腦組織,分別行肉眼觀察、HE切片染色、免疫組化,證實SCID小鼠膠質(zhì)瘤情況。結(jié)果:1.蒿甲醚可抑制人腦U251膠質(zhì)瘤細(xì)胞增殖,其抑瘤作用呈時間和劑量依賴性;蒿甲醚作用后的U251膠質(zhì)瘤細(xì)胞上清液中VEGF含量和生理鹽水組相比明顯降低,差異有統(tǒng)計學(xué)意義(P0.05)。2.成功構(gòu)建SCID小鼠U251人腦膠質(zhì)瘤原位模型,經(jīng)活體左心灌注后取腦組織發(fā)現(xiàn)均有腫瘤生長。3.蒿甲醚組和替莫唑胺組SCID小鼠的中位生存期較對照組有明顯延長(P0.05);腦組織HE切片染色可見對照組的U251細(xì)胞核大深染,呈梭型或不規(guī)則形,高倍鏡下可見U251細(xì)胞向正常腦組織浸潤,蒿甲醚組及替莫唑胺組腫瘤細(xì)胞密集程度均明顯低于對照組。結(jié)論:1、蒿甲醚可抑制人腦U251膠質(zhì)瘤細(xì)胞增殖,其抑瘤效應(yīng)呈劑量-時間依賴性。2、蒿甲醚可能通過抑制VEGF表達(dá)而發(fā)揮其抗腫瘤作用。3、蒿甲醚可抑制SCID小鼠顱內(nèi)膠質(zhì)瘤增殖。
[Abstract]:Objective: To observe the first effect of artemether on proliferation of U251 glioma cells in vitro, and then establish a SCID mouse glioma orthotopic model, observe the effect of artemether in vivo inhibition of glioma and its anti angiogenesis, investigate the mechanism of artemether inhibiting glioma proliferation, provides a new idea and method for the clinical treatment of glioma. Methods: Cultured U251 glioma cells in vitro 1. medium, using MTT (four methyl thiazolyl tetrazolium) method for the determination of inhibition effect of artemether on U251 glioma cell growth, and calculate the half inhibitory concentration (IC50); detection of artemether inhibition of U251 glioma cells by ELISA method the expression of VEGF.2. to construct SCID mouse glioma orthotopic model using stereotaxic apparatus positioning anesthesia mice, cut the scalp, from the sagittal suture and coronal suture respectively at the intersection of right back 2mm injection point positioning And after drilling, using 10ul micro syringe perpendicular to the skull in the mouse brain location (right parietal area) deep 5mm, injection of 10ulU251 human glioma cells, bone wax filling bone hole, absorbable suture scalp, living conditions in mice, epilepsy symptoms in mice after in vivo perfusion via the left ventricle after brain tissue visual observation, gross appearance and section, HE staining confirmed the tumor model of.3. is constructed successfully constructed 24 SCID mice U251 glioma orthotopic model, randomly divided into 3 groups, including control group (saline group (0.4ml/10g/d), artemether artemether 50.0mg/kg/d), temozolomide group (20mg/kg/d) temozolomide. Using the powder made of suspension gavage, weight of 1.65mg/ /d. after inoculation were labeled, regular daily dosing, the survival time was recorded. About 20 days after inoculation, when SCID mice died of left heart in vivo during reperfusion, take Out and fixed all of the brain, were observed by gross, HE staining, immunohistochemistry, confirmed SCID mouse glioma. Results: 1. artemether could inhibit the proliferation of human U251 glioma cells, the inhibitory effect was time and dose dependent effect of artemether; after U251 glioma cells supernatant the content of VEGF and saline group was significantly decreased, the difference was statistically significant (P0.05).2. was successfully constructed SCID mice U251 glioma orthotopic model by in vivo left ventricular perfusion after brain tissue found that the median survival time of tumor growth were.3. artemether group and temozolomide group in SCID mice than in the control group obviously extended (P0.05); brain tissue HE staining was observed in control group U251 nucleus anachromasis, spindle shaped or irregular shape, visible at high magnification U251 cell infiltration to normal brain tissue, artemether group and temozolomide group of tumor cell intensive process Conclusion: 1, artemether can inhibit the proliferation of human brain U251 glioma cells, and its anti-tumor effect is dose time dependent.2. Artemether can inhibit the expression of VEGF and exert its antitumor effect.3, artemether can inhibit the growth of intracranial glioma in SCID mice.

【學(xué)位授予單位】：昆明醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R739.41;R-332

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