本文選題：急性髓系白血病 + 益氣養(yǎng)陰方��； 參考：《山東中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的:以NOD/SCID小鼠為研究平臺(tái),以KG-1a細(xì)胞株建立人急性髓系白血病模型,探討益氣養(yǎng)陰方及其拆方通過(guò)線(xiàn)粒體途徑誘導(dǎo)急性髓系白血病KG-1a細(xì)胞凋亡的可能作用機(jī)理,為中醫(yī)藥通過(guò)線(xiàn)粒體途徑誘導(dǎo)白血病細(xì)胞凋亡為靶點(diǎn)治療急性髓系白血病尋求實(shí)驗(yàn)學(xué)支持,以期在此理論指導(dǎo)下研究新的抗白血病中藥提供理論依據(jù)。方法:以NOD/SCID小鼠為實(shí)驗(yàn)對(duì)象,經(jīng)60Co全身輻照,尾靜脈注射KG-1a細(xì)胞株,建立急性髓系白血病小鼠模型。將造模成功后小鼠隨機(jī)分為對(duì)照組、扶正組、祛邪組、全方組并以非造模小鼠為空白對(duì)照組作為陰性對(duì)照,觀(guān)察小鼠的一般生存狀態(tài),瀕死小鼠行骨髓系形態(tài)學(xué)及組織病理學(xué)檢測(cè),給藥4周后脫頸處死小鼠,取脾細(xì)胞制成細(xì)胞懸液應(yīng)用流式細(xì)胞術(shù)檢測(cè)NOD/SCID小鼠細(xì)胞周期與細(xì)胞凋亡率;采用免疫組化法檢測(cè)線(xiàn)粒體相關(guān)凋亡因子Cyt-C、Apaf-1、Smac/Diablo和AIF的表達(dá)。結(jié)果:藥物治療后,用藥組和空白對(duì)照組小鼠生存時(shí)間較對(duì)照組明顯延長(zhǎng)(P0.01),全方組和空白對(duì)照組小鼠生存時(shí)間較扶正組與祛邪組長(zhǎng)(P0.01),全方組小鼠較空白對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),祛邪組小鼠生存時(shí)間較扶正組延長(zhǎng)(P0.01)。用藥組細(xì)胞G1期停滯率和細(xì)胞凋亡率較對(duì)照組和空白對(duì)照組明顯升高(P0.01),全方組細(xì)胞G1期停滯率和細(xì)胞凋亡率較扶正組與祛邪組升高(P0.01),祛邪組細(xì)胞G1期停滯率較扶正組略高(P0.05),祛邪組細(xì)胞凋亡率較扶正組升高(P0.01)對(duì)照組與空白對(duì)照組比較無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。小鼠骨髓中Cyt-C、Apaf-1、Smac/Diablo、AIF的表達(dá)均為用藥組較對(duì)照組升高(P0.05),其中Cyt-C與Apaf-1表達(dá)顯著升高(P0.01);全方組均高于扶正組與祛邪組(P0.01);祛邪組均較扶正組升高(P0.05),其中Cyt-C、Apaf-1和Smac/Diablo表達(dá)顯著升高(P0.01)。其中對(duì)于各指標(biāo)的影響全方組均優(yōu)于扶正組與祛邪組,祛邪組均優(yōu)于扶正組。結(jié)論:實(shí)驗(yàn)結(jié)果表明,益氣養(yǎng)陰方能夠使停滯于G1期的細(xì)胞增加,進(jìn)入S期的細(xì)胞減少,從而了抑制白血病細(xì)胞增殖,并上調(diào)線(xiàn)粒體相關(guān)凋亡因子Cyt-C、Apaf-1、Smac/Diablo、AIF的表達(dá),誘導(dǎo)白血病細(xì)胞凋亡,提高白血病細(xì)胞的凋亡率,因此延長(zhǎng)了白血病小鼠生存期,揭示其對(duì)急性髓系白血病具有一定的治療作用。
[Abstract]:Objective: to establish human acute myeloid leukemia model with KG-1a cell line and to explore the possible mechanism of inducing apoptosis of KG-1a cells by mitochondrial pathway. The aim of this study is to seek experimental support for the treatment of acute myeloid leukemia by inducing apoptosis of leukemic cells through mitochondrial pathway in order to provide theoretical basis for the study of new anti-leukemia Chinese medicine under the guidance of this theory. Methods: the mouse model of acute myeloid leukemia was established by injecting KG-1a cell line into caudal vein after 60Co irradiation. The mice were randomly divided into three groups: control group, Fuzheng group, dispelling evil group, the whole prescription group and the non-model mice as the negative control group. The general living state of the mice was observed. Bone marrow morphology and histopathological examination were performed in the dying mice. The mice were killed after 4 weeks of administration. The splenocytes were made into cell suspensions and the cell cycle and apoptosis rate of nod / SCID mice were detected by flow cytometry. The expression of mitochondrial related apoptosis factor Cyt-Capaf-1 Smac-Diablo and AIF was detected by immunohistochemical method. Results: after drug treatment, The survival time of mice in the medication group and blank control group was significantly longer than that in the control group (P0.01). The survival time of mice in the whole prescription group and blank control group was significantly longer than that in the Fuzheng group and in the blank control group (P0.01), but there was no statistical significance in the whole prescription group compared with the blank control group (P0.05). The survival time of mice in evil group was longer than that in Fuzheng group (P0.01). Compared with the control group and the blank control group, the G1 phase arrest rate and cell apoptosis rate of the medication group were significantly higher than those of the control group and the blank control group (P0.01). The G1 phase arrest rate and the apoptosis rate of the whole prescription group were higher than those of the Fuzheng group and the dispelling evil group (P0.01). The positive group was slightly higher (P0.05), and the apoptosis rate of the pathogenic group was higher than that of the Fuzheng group (P0.01). There was no significant difference between the control group and the blank control group (P0.05). The expressions of Cyt-C and Apaf-1 in bone marrow of mice were higher than those of control group (P0.05), the expression of Cyt-C and Apaf-1 were significantly higher in the whole prescription group than in the Fuzheng group and dispelling evil group (P0.01), and the expression of Cyt-Capaf-1 and Smac-Diablo in the pathogenic group were significantly higher than those in the Fuzheng group (P0.05), and the expression of Cyt-Capaf-1 and Smac-Diablo in the treatment group were significantly higher than that in the control group (P0.01). The effect on each index in the whole prescription group was better than that in the Fuzheng group and the dispelling evil group, and that in the dispelling evil group was better than that in the Fuzheng group. Conclusion: the experimental results showed that Yiqi Yangyin prescription could increase the number of cells in G1 phase and decrease the number of cells in S phase, thereby inhibiting the proliferation of leukemia cells and up-regulating the expression of Cyt-Cnapaf-1, a mitochondrial related apoptosis factor, in Smac-P / Diablosi-AIF. The apoptosis of leukemia cells was induced and the apoptosis rate of leukemia cells was increased. Therefore, the survival time of leukemia mice was prolonged, and it was revealed that it had a certain therapeutic effect on acute myeloid leukemia.
【學(xué)位授予單位】：山東中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R273

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