RDP-p53融合蛋白对小鼠炎症细胞因子的影响
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摘要
研究不同剂量RDP-p53融合蛋白对小鼠炎症细胞因子的影响。通过大肠杆菌Rosetta表达蛋白并纯化,SDS-PAGE确定RDP-p53融合蛋白准确性。同时,将昆明小鼠分为空白对照组和RDP-p53融合蛋白高、中、低剂量组,经腹腔注射给药,摘眼球取血,用酶联免疫吸附法(ELISA)测定血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的含量。RDP-p53融合蛋白在上清和沉淀中均获得表达。ELISA测定结果表明,与对照组比较,低、中剂量组小鼠血清炎症因子水平没有明显变化(P>0.05),高剂量组的IL-1β和TNF-α含量显著升高(P<0.05)。成功制备RDP-P53融合蛋白,并为进一步研究RDP-p53融合蛋白抗神经胶质瘤药理作用时给药剂量的确定提供了实验依据。
The effects of different doses of RDP-p53 fusion protein in treatment mice's inflammatory cytokine is studied. RDP-p53 fusion protein was expressed by E. coli Rosetta and got purified,the expressed product was identified by SDS-PAGE. Then Kunming mice were divided into 4 groups,i. e. blank control group and RDP-p53 fusion protein high-dose,medium-dose and low-dose groups. Kunming mice were immunized by peritoneal route,of which the sera were separated and determined for IL-1β,IL-6 and TNF-α level by ELISA. RDP-p53 fusion protein got expression in both supernatant and precipitation. ELISA results showed that: compared with control group,inflammatory cytokine content of low-dose and medium-dose groups didn't increase significantly( P>0. 05),IL-1βand TNF-α content of high-dose groups increased significantly( P<0. 05). RDP-p53 fusion protein was successfully prepared,this might provide some valuable information for determining dosage of administration while studying pharmacological effect that RDP-p53 fusion protein inhibits brain tumors.
The effects of different doses of RDP-p53 fusion protein in treatment mice's inflammatory cytokine is studied. RDP-p53 fusion protein was expressed by E. coli Rosetta and got purified,the expressed product was identified by SDS-PAGE. Then Kunming mice were divided into 4 groups,i. e. blank control group and RDP-p53 fusion protein high-dose,medium-dose and low-dose groups. Kunming mice were immunized by peritoneal route,of which the sera were separated and determined for IL-1β,IL-6 and TNF-α level by ELISA. RDP-p53 fusion protein got expression in both supernatant and precipitation. ELISA results showed that: compared with control group,inflammatory cytokine content of low-dose and medium-dose groups didn't increase significantly( P>0. 05),IL-1βand TNF-α content of high-dose groups increased significantly( P<0. 05). RDP-p53 fusion protein was successfully prepared,this might provide some valuable information for determining dosage of administration while studying pharmacological effect that RDP-p53 fusion protein inhibits brain tumors.
引文
1 汪进良,焦顺昌.p53基础研究及药物应用新进展.中国药物应用与监测,2008;5(2):38—40
2 Kumar P,Wu H,McBride J L,et al.Transvascular delivery of small interfering RNA to the central nervous system.Nature,2007;448(7149):39—43
3 Fu A,Zhao Z,Zhang M,et al.Cellular uptake mechanism and therapeutic utility of a novel peptide in targeted-delivery of protrins into neuronal cells.Pharm Res,2013;30(8):2108—2117
4 严世荣,王立林,邱方城.细胞穿膜肽的研究进展.生物技术通讯,2006;17(5):796—798
5 马晓丽,许士国,孙海基.COX-2和p53在炎症应激和肿瘤发生中相互作用的研究进展.国际遗传学杂志,2009;32(5):372 —376
6 薛秀霞.治疗用生物大分子免疫原性的研究进展.齐鲁药事,2012;31(3):174—178
7 曾行,毕惠嫦,黄明.炎症反应过程中药物转运体调控的研究进展.药学学报,2011;46(7):773—779
8 唐威华,张景六,王宗阳,等.SDS-PAGE法测定His-tag融合蛋白分子量产生偏差的原因.植物生理学报,2000;26(1):64—68
9 Yu Z,Wu J,Wu S,et al.A recombinant cell-permeable p53 fusion protein is selectively stabilized under hypoxia and inhibits tumor cell growth.Cancer Letters,2009;279(1):101—107
10 庄春林,缪震元,盛春泉,等.干扰p53-MDM2蛋白相互作用的小分子抑制剂的研究进展.中国药物化学杂志,2010;20(5):403 —407
11 王逸麟,付爱玲.狂犬病毒糖蛋白衍生肽作为靶脑载体初探.科学技术与工程,2011;11(36):8974—8977
12 付爱玲,赵宝全.蛋白质和核酸的入脑转运.重庆:西南师范大学出版社,2012:124
13 张晶,钟武,李松.TNF-α抑制剂的研究进展.中国药物化学杂志,2010;20(4):310—318
14 Kirkham B W,Lassere M N,Edmonds J P,et al.Synovial membrane cytokine expression is predictive of joint damage progression in rheumatoid arthritis:a two-year prospective study(the DAMAGE study cohort).Arthritis Rheum,2006;54(4):1122—1231
2 Kumar P,Wu H,McBride J L,et al.Transvascular delivery of small interfering RNA to the central nervous system.Nature,2007;448(7149):39—43
3 Fu A,Zhao Z,Zhang M,et al.Cellular uptake mechanism and therapeutic utility of a novel peptide in targeted-delivery of protrins into neuronal cells.Pharm Res,2013;30(8):2108—2117
4 严世荣,王立林,邱方城.细胞穿膜肽的研究进展.生物技术通讯,2006;17(5):796—798
5 马晓丽,许士国,孙海基.COX-2和p53在炎症应激和肿瘤发生中相互作用的研究进展.国际遗传学杂志,2009;32(5):372 —376
6 薛秀霞.治疗用生物大分子免疫原性的研究进展.齐鲁药事,2012;31(3):174—178
7 曾行,毕惠嫦,黄明.炎症反应过程中药物转运体调控的研究进展.药学学报,2011;46(7):773—779
8 唐威华,张景六,王宗阳,等.SDS-PAGE法测定His-tag融合蛋白分子量产生偏差的原因.植物生理学报,2000;26(1):64—68
9 Yu Z,Wu J,Wu S,et al.A recombinant cell-permeable p53 fusion protein is selectively stabilized under hypoxia and inhibits tumor cell growth.Cancer Letters,2009;279(1):101—107
10 庄春林,缪震元,盛春泉,等.干扰p53-MDM2蛋白相互作用的小分子抑制剂的研究进展.中国药物化学杂志,2010;20(5):403 —407
11 王逸麟,付爱玲.狂犬病毒糖蛋白衍生肽作为靶脑载体初探.科学技术与工程,2011;11(36):8974—8977
12 付爱玲,赵宝全.蛋白质和核酸的入脑转运.重庆:西南师范大学出版社,2012:124
13 张晶,钟武,李松.TNF-α抑制剂的研究进展.中国药物化学杂志,2010;20(4):310—318
14 Kirkham B W,Lassere M N,Edmonds J P,et al.Synovial membrane cytokine expression is predictive of joint damage progression in rheumatoid arthritis:a two-year prospective study(the DAMAGE study cohort).Arthritis Rheum,2006;54(4):1122—1231