本文選題：胰腺癌　切入點：黃芩素　出處：《南京中醫(yī)藥大學》2017年博士論文

【摘要】：胰腺癌是惡性程度高、死亡率高的消化系統(tǒng)腫瘤,由于胰腺癌的診斷困難,發(fā)病早期癥狀不明顯,目前對胰腺癌的治療仍不理想,生存率不足5%。目前吉西他濱是胰腺癌輔助化療的一線藥物,但對胰腺癌患者生存率的提高仍然不足。近年來,多個臨床試驗將吉西他濱和其他藥物聯(lián)合治療胰腺癌,但是效果仍不理想。因此,尋找更有效的治療胰腺癌的藥物,或者能聯(lián)合吉西他濱達到更好治療效果的藥物十分重要。中藥黃芩的主要有效成分是黃芩素,黃芩素也是黃芩主要發(fā)揮藥理作用的成分。研究發(fā)現(xiàn),黃芩素具有多種藥理學作用,特別對多種腫瘤具有顯著的抑制效果。小劑量短時間經(jīng)過黃芩素的處理,腫瘤細胞能得到有效抑制,而當大劑量長時間黃芩素處理后,能誘導腫瘤細胞凋亡。因此,我們考慮通過小劑量的黃芩素聯(lián)合吉西他濱是否能有效的抑制胰腺癌細胞的生長,成為潛在的治療手段。研究方法(1)體外研究黃芩素聯(lián)合吉西他濱對胰腺癌細胞的抑制作用及機制1、MTT檢測黃芩素、吉西他濱和兩種藥物聯(lián)用對胰腺癌細胞系CFPAC-1增殖的抑制作用;2、Hoechst33258染色,熒光顯微鏡下觀察黃芩素及黃芩素聯(lián)合吉西他濱對胰腺癌細胞的影響;3、Annexin-V APC/7-AAD雙染法檢測黃芩素、吉西他濱和兩種藥物聯(lián)用對胰腺癌細胞凋亡的影響;4、黃芩素、吉西他濱和兩種藥物聯(lián)用在胰腺癌細胞中對凋亡相關(guān)蛋白表達的影響;(2)藥物治療胰腺癌裸鼠移植模型將人胰腺癌細胞系CFPAC-1注射到裸鼠背部建立裸鼠移植瘤模型,待皮下腫瘤生長到2-4 mm開始給藥,分別給予黃芩素、吉西他濱和兩種藥物聯(lián)用,記錄裸鼠體重及皮下移植瘤體積,共觀察28天。然后處死裸鼠,取腫瘤進行稱重和免疫組化染色。實驗結(jié)果1、黃芩素或吉西他濱均能顯著抑制人胰腺癌細胞系CFPAC-1細胞增殖,并且具有藥物濃度依耐性,并且兩種藥物聯(lián)用對細胞增殖抑制效果更為顯著;2、黃芩素與吉西他濱聯(lián)用能誘導人胰腺癌細胞系CFPAC-1細胞凋亡;3、黃芩素與吉西他濱聯(lián)用誘導胰腺癌細胞凋亡是通過調(diào)控凋亡蛋白表達改變起作用的;4、黃芩素與吉西他濱聯(lián)用能顯著抑制人胰腺癌細胞裸鼠移植瘤的生長,并且作用效果明顯優(yōu)于單藥的作用。結(jié)論黃芩素可以抑制胰腺癌細胞的增殖,誘導其分化。體內(nèi)外實驗研究表明,聯(lián)合黃芩素和吉西他濱可以明顯抑制人胰腺癌細胞的增殖,效果優(yōu)于單藥處理。黃芩素和吉西他濱聯(lián)用將是成為胰腺癌治療的新手段。
[Abstract]:Pancreatic cancer is a kind of digestive system tumor with high malignancy and high mortality. Because of the difficulty in diagnosis of pancreatic cancer, the early symptoms of pancreatic cancer are not obvious, so the treatment of pancreatic cancer is still not ideal, and the survival rate is less than 5%.Gemcitabine is the first-line adjuvant chemotherapy for pancreatic cancer, but the improvement of survival rate is still insufficient.In recent years, several clinical trials have combined gemcitabine with other drugs to treat pancreatic cancer, but the results are still unsatisfactory.Therefore, it is important to find more effective drugs for pancreatic cancer, or combination of gemcitabine for better treatment.The main active component of Scutellaria baicalensis is baicalin, and baicalin is the main pharmacological component of Scutellaria baicalensis.It was found that baicalin has many pharmacological effects, especially on many tumors.After the treatment of baicalin for a short time, the tumor cells could be effectively inhibited, but when the large dose of baicalin was treated for a long time, the apoptosis of tumor cells could be induced.Therefore, we consider whether small doses of baicalin combined with gemcitabine can effectively inhibit the growth of pancreatic cancer cells as a potential treatment.Methods 1) to study the inhibitory effect of baicalin combined with gemcitabine on pancreatic cancer cells in vitro and its mechanism. The inhibitory effects of baicalin, gemcitabine and two drugs on the proliferation of pancreatic cancer cell line CFPAC-1 were detected by MTT assay.The effects of baicalin and combination of baicalin and gemcitabine on pancreatic cancer cells were observed under fluorescence microscope. The effects of baicalin, gemcitabine and two drugs on apoptosis of pancreatic cancer cells were detected by Annexin-V APC/7-AAD double staining.Effects of gemcitabine and two drugs on the expression of apoptosis-related protein in pancreatic cancer cellsBaicalin, gemcitabine and two drugs were used to record the body weight of nude mice and the volume of subcutaneous transplanted tumor, and the tumor volume was observed for 28 days after the subcutaneous tumor grew to 2-4 mm and was given baicalin, gemcitabine and two kinds of drugs respectively.Then the nude mice were killed and the tumor was weighed and stained by immunohistochemistry.Results 1. Baicalin or gemcitabine could significantly inhibit the proliferation of human pancreatic cancer cell line CFPAC-1, and had concentration-dependent drug tolerance.The combination of baicalin and gemcitabine can induce apoptosis of human pancreatic cancer cell line CFPAC-1, baicalin and gemcitabine can induce apoptosis of pancreatic cancer cell line through the combination of baicalin and gemcitabine.Regulating the expression of apoptotic protein and regulating the expression of apoptotic protein, baicalin combined with gemcitabine could significantly inhibit the growth of human pancreatic cancer cell xenografts in nude mice.And the effect was obviously better than that of single drug.Conclusion baicalin can inhibit the proliferation and induce the differentiation of pancreatic cancer cells.In vitro and in vivo experiments showed that the combination of baicalin and gemcitabine could significantly inhibit the proliferation of human pancreatic cancer cells.Baicalin combined with gemcitabine will be a new treatment for pancreatic cancer.
【學位授予單位】：南京中醫(yī)藥大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：R735.9

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