紫草素对荷肝癌裸小鼠移植瘤生长的抑制作用及机制
|
摘要
目的研究紫草素抑制荷肝癌裸小鼠移植瘤生长的作用及其机制。方法通过皮下注射肝癌细胞制备荷肝癌裸小鼠模型;设模型组,紫草素低、中、高剂量组和顺铂组,每组12只,分别隔天一次给予0.9%氯化钠溶液,紫草素0.5mg/kg、1mg/kg、2mg/kg和顺铂2 mg/kg,共给药6次。观察动物生存状态和移植瘤组织生长状态,称量瘤重并计算抑瘤率;HE染色法进行肿瘤组织形态检查,TUNEL法进行细胞凋亡检查,免疫组织化学法观察肿瘤组织凋亡相关蛋白表达。结果与模型组比较,紫草素各组裸小鼠食欲和精神状态都明显好转,瘤体积小、活动度低、硬度低,移植瘤组织出现片状坏死病变、移植瘤组织细胞凋亡数量明显增多,凋亡指数显著升高[(22.93±5.40)、(39.86±8.25)、(60.74±9.61)比(2.64±1.09),P<0.05];瘤体质量显著减轻[(1.32±0.21)g、(1.18±0.17)g、(0.83±0.12)g比(1.69±0.14)g,P<0.05]、抑瘤率显著升高[(21.89±3.75)%、(30.18±4.92)%、(50.89±6.43)%比(0.00±0.00)%,P<0.01];肿瘤组织中凋亡相关蛋白[半胱氨酸天冬氨酸蛋白酶3(caspase-3)、bcl-2相关X蛋白质(Bax、bcl-2)]中的caspase-3表达显著上调[(0.16±0.05)、(0.21±0.04)、(0.27±0.06)比(0.05±0.02),P<0.05]、Bax表达显著上调[(0.54±0.10)、(0.63±0.15)、(0.81±0.21)比(0.29±0.05),P<0.01],其中紫草素中、高剂量组bcl-2显著下调[(0.37±0.05)、(0.30±0.05)比(0.50±0.07),P<0.05],Bax/bcl-2比值显著升高[(1.17±0.25)、(1.70±0.39)、(2.71±0.68)比(0.59±0.20),P<0.05]。结论紫草素具有抑制荷肝癌裸小鼠移植瘤生长的作用,可能与其调节凋亡相关蛋白表达而促进细胞凋亡、诱导肿瘤组织坏死有关。
Objective To investigate the effect of Shikonin on the growth of transplanted tumor in nude mice bearing liver cancer and its mechanism. Methods The nude mice models bearing liver cancer were made by subcutaneous injection of human liver cell line HepG2, and were treated with Shikonin(0.5.1,2 mg/kg) and Cisplatin(2 mg/kg),once every two days,for 6 times.The mice ' general states and tumor growth conditions were observed, the histopathological changes of tumor tissue was observed by HE staining;the tumor cells apoptosis was detected by TUNEL;the expression of apoptosis-related protein were detected by immunohistochemistry(IHC).Results After treated with Shikonin,the diet and mental state of nude mice bearing liver cancer were improved, tumor size, activity and hardness decreased; thetumor tissue showed pathological changes such as necrosis, cell shrinkage,the number of apoptotic cells in tumor tissue increased significantly,and the apoptotic index increased significantly[(22.93±5.40),(39.86±8.25),(60.74±9.61)vs.(2.64± 1.09),P< 0.05 ]; the tumor,weight of Shikonin treated groups decreased significantly[(1.32±0.21) g,(1.18±0. 17) g,(0.83±0.12) g vs.(1.69±0.14)g,P < 0.05 or P < 0.01 ]; the inhibition rate increased significantly[(21.89±3.75) %,(30.18±4.92) %,(50.89±6.43) %vs.(0.00±0.00)%,P < 0.01]; the expression of Bax [(0.16±0.05),(0.21±0.04),(0.27±0.06)vs.(0.05±0.02),P< 0.05] and caspase-3 [(0.54±0.10),(0.63±0.15),(0.81 ±0.21) vs.(0.29±0.05),P< 0.01 ] increased significantly, the expression of Bcl-2 protein in Shikonin.(1,2 mg/kg) groups decreased significantly [(0.37±0.05),(0.30±0.05)vs.(0.50±0.07),P < 0.05],and the ratio of Bax/Bcl-2 were increased significantly [(1.17±0.25),(1.70±0.39),(2.71±0.68)vs.(0.59±0.20), P < 0.05 ].Conclusions Shikonin inhibits the growth of xenografted liver cancer in nude mice, which may be related to the regulation of apoptosis-related protein expression, apoptosis and tumor necrosis.
Objective To investigate the effect of Shikonin on the growth of transplanted tumor in nude mice bearing liver cancer and its mechanism. Methods The nude mice models bearing liver cancer were made by subcutaneous injection of human liver cell line HepG2, and were treated with Shikonin(0.5.1,2 mg/kg) and Cisplatin(2 mg/kg),once every two days,for 6 times.The mice ' general states and tumor growth conditions were observed, the histopathological changes of tumor tissue was observed by HE staining;the tumor cells apoptosis was detected by TUNEL;the expression of apoptosis-related protein were detected by immunohistochemistry(IHC).Results After treated with Shikonin,the diet and mental state of nude mice bearing liver cancer were improved, tumor size, activity and hardness decreased; thetumor tissue showed pathological changes such as necrosis, cell shrinkage,the number of apoptotic cells in tumor tissue increased significantly,and the apoptotic index increased significantly[(22.93±5.40),(39.86±8.25),(60.74±9.61)vs.(2.64± 1.09),P< 0.05 ]; the tumor,weight of Shikonin treated groups decreased significantly[(1.32±0.21) g,(1.18±0. 17) g,(0.83±0.12) g vs.(1.69±0.14)g,P < 0.05 or P < 0.01 ]; the inhibition rate increased significantly[(21.89±3.75) %,(30.18±4.92) %,(50.89±6.43) %vs.(0.00±0.00)%,P < 0.01]; the expression of Bax [(0.16±0.05),(0.21±0.04),(0.27±0.06)vs.(0.05±0.02),P< 0.05] and caspase-3 [(0.54±0.10),(0.63±0.15),(0.81 ±0.21) vs.(0.29±0.05),P< 0.01 ] increased significantly, the expression of Bcl-2 protein in Shikonin.(1,2 mg/kg) groups decreased significantly [(0.37±0.05),(0.30±0.05)vs.(0.50±0.07),P < 0.05],and the ratio of Bax/Bcl-2 were increased significantly [(1.17±0.25),(1.70±0.39),(2.71±0.68)vs.(0.59±0.20), P < 0.05 ].Conclusions Shikonin inhibits the growth of xenografted liver cancer in nude mice, which may be related to the regulation of apoptosis-related protein expression, apoptosis and tumor necrosis.
引文
[1]张永红,于慧敏,聂英坤.紫草素对类风湿关节炎滑膜成纤维细胞COX-2表达的影响[J].哈尔滨医科大学学报,2012,46(3):261-265.
[2]代巧妹,李冀,颜培宇.紫草素免疫调节作用研究进展[J].中医药信息,2012,29(6):116-118.
[3]陈菊英,刘朝纯,曾智,等.紫草素通过PI3K/Akt通路促进人乳腺癌MCF-7细胞自噬[J].中国药理学通报,2013,29(2):194-198.
[4]杨京哲,张凌霄,邢永静,等.紫草素对子宫内膜癌Ishikawa细胞生长的作用研究[J].现代中西医结合杂志,2013,22(23):2520-2523.
[5]于海荣,苗浩,庞冲,等.紫草素对人宫颈癌Hela细胞增殖的影响及其作用机制研究[J].中国生化药物杂志,2015,35(4):16-18.
[6]鲁昊,康健.紫草素对人喉癌Hep-2细胞生长的作用研究[J].东南大学学报(医学版),2011,30(2):350-354.
[7]陈志炉,戴其舟,王勇,等.紫草素对白血病细胞HL-60增殖及凋亡作用的影响[J].中国中西医结合杂志,2012,32(2):239-243.
[8]谢丹,林源,杨阿莉.大黄素抑制人肝癌裸鼠移植瘤生长机制探讨[J].中华肿瘤防治杂志,2015,22(1):28-33.
[9]高姗,杨万水,高静,等.原发性肝癌的分子流行病学研究进展[J].中国肿瘤,2012,21(2):136-144.
[10]吕桂帅,陈磊,王红阳.我国肝癌研究的现状与前景[J].生命科学,2015,27(3):237-248.
[11] ANSARI D,ANDERSSON R.Radiofrequency ablation or percuta-neous ethanol injection for the treatment of liver tumors[J].World J Gastroenterol,2012,18(10):1003-1008.
[12] FARHADI F,JAHANPOUR S,HAZEM K,et al.Garlic(Allium sativum)fresh juice induces apoptosis in human oral squamous cell carcinoma:the involvement of caspase-3,bax and bcl-2[J].J Dent Res Dent Clin Dent Prospects,2015,9(4):267-273.
[13] OGURAT,TANAKAY,TAMAKIH,etal.Docetaxelinduces Bcl-2-and pro-apoptotis caspase-independent death of human prostate cancer DU145 cells[J].Int J Oncol,2016,48(6):2330-2338.
[14]焦卫云,陈道俊,柯章敏,等.姜黄素增加U937细胞对TRAIL的敏感性的凋亡机制研究[J].安徽医药,2015,19(3):563-565.
[15]叶记林,于有江,吴爱莲,等.氧化应激在TRAIL诱导Hela细胞凋亡中的作用[J].安徽医药,2015,19(11):2090-2093.
[16] SAEEDI BORUJENI MJ,HAMI J,HAGHIR H,et al.Evaluation of bax and bcl-2 proteins expression in the rat hippocampus due to childhoodfebrileseizure[J].IranJChildNeurol,2016,10(1):53-60.
[2]代巧妹,李冀,颜培宇.紫草素免疫调节作用研究进展[J].中医药信息,2012,29(6):116-118.
[3]陈菊英,刘朝纯,曾智,等.紫草素通过PI3K/Akt通路促进人乳腺癌MCF-7细胞自噬[J].中国药理学通报,2013,29(2):194-198.
[4]杨京哲,张凌霄,邢永静,等.紫草素对子宫内膜癌Ishikawa细胞生长的作用研究[J].现代中西医结合杂志,2013,22(23):2520-2523.
[5]于海荣,苗浩,庞冲,等.紫草素对人宫颈癌Hela细胞增殖的影响及其作用机制研究[J].中国生化药物杂志,2015,35(4):16-18.
[6]鲁昊,康健.紫草素对人喉癌Hep-2细胞生长的作用研究[J].东南大学学报(医学版),2011,30(2):350-354.
[7]陈志炉,戴其舟,王勇,等.紫草素对白血病细胞HL-60增殖及凋亡作用的影响[J].中国中西医结合杂志,2012,32(2):239-243.
[8]谢丹,林源,杨阿莉.大黄素抑制人肝癌裸鼠移植瘤生长机制探讨[J].中华肿瘤防治杂志,2015,22(1):28-33.
[9]高姗,杨万水,高静,等.原发性肝癌的分子流行病学研究进展[J].中国肿瘤,2012,21(2):136-144.
[10]吕桂帅,陈磊,王红阳.我国肝癌研究的现状与前景[J].生命科学,2015,27(3):237-248.
[11] ANSARI D,ANDERSSON R.Radiofrequency ablation or percuta-neous ethanol injection for the treatment of liver tumors[J].World J Gastroenterol,2012,18(10):1003-1008.
[12] FARHADI F,JAHANPOUR S,HAZEM K,et al.Garlic(Allium sativum)fresh juice induces apoptosis in human oral squamous cell carcinoma:the involvement of caspase-3,bax and bcl-2[J].J Dent Res Dent Clin Dent Prospects,2015,9(4):267-273.
[13] OGURAT,TANAKAY,TAMAKIH,etal.Docetaxelinduces Bcl-2-and pro-apoptotis caspase-independent death of human prostate cancer DU145 cells[J].Int J Oncol,2016,48(6):2330-2338.
[14]焦卫云,陈道俊,柯章敏,等.姜黄素增加U937细胞对TRAIL的敏感性的凋亡机制研究[J].安徽医药,2015,19(3):563-565.
[15]叶记林,于有江,吴爱莲,等.氧化应激在TRAIL诱导Hela细胞凋亡中的作用[J].安徽医药,2015,19(11):2090-2093.
[16] SAEEDI BORUJENI MJ,HAMI J,HAGHIR H,et al.Evaluation of bax and bcl-2 proteins expression in the rat hippocampus due to childhoodfebrileseizure[J].IranJChildNeurol,2016,10(1):53-60.