本文選題：肝細(xì)胞癌 + 活血化瘀中藥��； 參考：《成都中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的：本實(shí)驗(yàn)選用臨床上被廣泛用于惡性腫瘤治療的紅花、桃仁、懷牛膝等具有活血化瘀功效的中藥,基于TG品系flila-EGFP轉(zhuǎn)基因斑馬魚生物模型來篩選其中具有抑制腫瘤新生血管生成效用的中藥,客觀、系統(tǒng)地評價(jià)紅花、桃仁、懷牛膝等具有活血化瘀功效的中藥在臨床上應(yīng)用于治療惡性腫瘤的可靠性和安全性,為臨床使用活血化瘀中藥治療惡性腫瘤提供客觀的實(shí)驗(yàn)依據(jù)。方法：選用TG品系的flila-EGFP轉(zhuǎn)基因斑馬魚為生物模型,每個(gè)樣品處理30只斑馬魚胚胎,在12孔板(Falcon)中實(shí)驗(yàn)。將工作濃度為200μg/ml的紅花、桃仁、懷牛膝水提取物和陽性對照藥物-PTK787 (5 μ g/ml)分別處理受精后22小時(shí)(22hpf)的斑馬魚胚胎直至48hpf。完整的連接背主動脈(DA)和背長軸血管(DLAV)之間的一段血管,即體節(jié)間血管(ISV),是衡量斑馬魚血管新生(angiogenesis)的重要指標(biāo)。藥物處理實(shí)驗(yàn)結(jié)束后,在體視顯微鏡(明場)和體視熒光顯微鏡下進(jìn)行血管表型觀察和拍照,拍照時(shí)相為48hpf,以分析各個(gè)樣品對斑馬魚體節(jié)間血管(ISV)生成的影響。同時(shí),觀察和統(tǒng)計(jì)各個(gè)樣品誘導(dǎo)的斑馬魚發(fā)育毒性和死亡數(shù)。結(jié)果：各個(gè)樣品均未出現(xiàn)藥物誘導(dǎo)的斑馬魚發(fā)育毒性和死亡數(shù)事件。體視顯微鏡(明場)和體視熒光顯微鏡下完整的ISVs表型觀察顯示：空白對照組斑馬魚胚胎的ISVs的表達(dá)完整、清晰；與空白對照組相比較,陽性對照組采用瓦他拉尼干預(yù)的斑馬魚胚胎的ISV及DLAV的熒光表達(dá)缺失；紅花、桃仁、懷牛膝水提物組斑馬魚胚胎的ISVs的熒光表達(dá)都有不同程度的變細(xì)、熒光強(qiáng)度減弱,或分布紊亂；其中,懷牛膝、紅花水提物組斑馬魚胚胎ISVs的熒光表達(dá)變細(xì)、強(qiáng)度減弱較為明顯,且分布紊亂,懷牛膝水提物組尤為突出。結(jié)論：瓦他拉尼能完全抑制斑馬魚胚胎的ISVs及DLAV;臨床治療腫瘤常用活血化瘀中藥紅花、桃仁、懷牛膝對斑馬魚胚胎的ISVs有不同程度的抑制作用；活血化瘀中藥紅花、桃仁、懷牛膝對轉(zhuǎn)基因斑馬魚新生血管生成存在抑制效用,從而抑制腫瘤生長及轉(zhuǎn)移；活血化瘀中藥懷牛膝、紅花、桃仁抑制腫瘤新生血管生成效應(yīng)有差異,其中,懷牛膝、紅花作用明顯,且安全、可靠,無致畸、致死事件；推測腫瘤活血化瘀療法可以抑制腫瘤血管生成,從而抑制腫瘤生長及轉(zhuǎn)移。
[Abstract]:Objective: to select traditional Chinese medicine which is widely used in the treatment of malignant tumor, such as safflower, peach kernel and Achyranthes bidentata, which has the effect of promoting blood circulation and removing blood stasis. Based on the TG flila-EGFP transgenic zebrafish biological model to screen the Chinese medicine which can inhibit tumor angiogenesis, objective, systematic evaluation of safflower, peach kernel, The traditional Chinese medicine with the effect of activating blood circulation and removing blood stasis is applied to the reliability and safety of malignant tumor in clinic, which provides an objective experimental basis for the clinical use of traditional Chinese medicine for promoting blood circulation and removing stasis in the treatment of malignant tumor. Methods: Flila-EGFP transgenic zebrafish from TG strain was used as a biological model. 30 zebrafish embryos were treated with each sample. Zebrafish embryos with working concentration of 200 渭 g/ml of safflower, peach kernel, Achyranthes bidentata and positive control drug -PTK787 5 渭 g / ml were treated for 22 hours after fertilization until 48hpf. The complete connection between the dorsal aorta (DAA) and the dorsal long axis vessel (DLAV) is an important indicator of angiogenesis in zebrafish. After the drug treatment, the phenotype of blood vessels was observed and photographed under stereoscopic microscope and stereoscopic fluorescence microscope, and the phase was 48hpf. to analyze the effect of each sample on the formation of ISV in zebrafish. At the same time, the developmental toxicity and death number of zebrafish induced by each sample were observed and counted. Results: there were no developmental toxicity and death events induced by drugs in all samples. The complete phenotypic observation of ISVs under stereoscopic microscope and stereoscopic fluorescence microscope showed that the expression of ISVs in zebrafish embryos in blank control group was complete and clear, and compared with that in blank control group. In the positive control group, the fluorescence expression of ISV and DLAV was absent in zebrafish embryos treated with Vataranil, and the fluorescence expression of ISVs of zebrafish embryos in safflower, peach kernel and Achyranthes bidentata water extract groups were all attenuated in varying degrees. The fluorescence expression of ISVs in zebrafish embryos became thinner, the intensity weakened obviously, and the distribution was disordered, especially in the water extract group of Achyranthes bidentata. Conclusion: Vataranil can completely inhibit the ISVs and DLAV of zebrafish embryos, the traditional Chinese medicine safflower, peach kernel and Achyranthes bidentata can inhibit the ISVs of zebrafish embryos in different degrees, and the traditional Chinese medicine safflower and peach kernel for activating blood circulation and removing stasis can inhibit the ISVs of zebrafish embryos in different degrees. The inhibitory effect of Achyranthes bidentata on the angiogenesis of transgenic zebrafish, thus inhibiting tumor growth and metastasis, is different from that of traditional Chinese medicine for promoting blood circulation and removing blood stasis, such as Achyranthes bidentata, safflower and peach kernel, among which, Achyranthes bidentata. The effect of safflower is obvious, safe, reliable, no teratogenicity, lethal event. It is inferred that tumor blood circulation and blood stasis therapy can inhibit tumor angiogenesis, thus inhibit tumor growth and metastasis.
【學(xué)位授予單位】：成都中醫(yī)藥大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：R285

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