TGFα与bFGF联结皂草毒素蛋白对增殖平滑肌细胞和内皮细胞的特异性细胞毒作用  

Cytotoxic effects of TGFα and bFGF conjugated with cytotoxin saporin on smooth musclec cells and endothelial cells

作　　者：杨军[1] 褚春[2] 周先令[1] 全智华[1] 余细勇[3] 林曙光[3] 

机构地区：[1]南华大学附一医院心内科,湖南衡阳421001 [2]南华大学附二医院药剂科,421001 [3]广东省心血管病研究所,广州510120

出　　处：《国际心血管病杂志》2010年第4期242-245,共4页International Journal of Cardiovascular Disease

基　　金：湖南省卫生厅重点课题(2001-Z06)

摘　　要：目的:为比较生物导向药物转化生长因子-皂草毒素蛋白(TGFα-SAP)和碱性成纤维细胞生长因子(bFGF)-SAP对增殖平滑肌细胞特异性细胞毒性作用。方法:用SPDP化学联结的方法合成了导向药物TGFα-SAP和bFGF-SAP,以3H-亮氨酸渗入法观察了TGFα-SAP及bFGF-SAP对平滑肌细胞(SMCs)和内皮细胞(ECs)蛋白质合成的影响,并以MTS法和3H-胸腺嘧啶核苷(TdR)渗入法比较TGFα-SAP及bFGF-SAP对SMCs和ECs的增殖和DNA合成的抑制作用。结果:10-9mol/L的TGFα-SAP可明显抑制SMCs增殖,SMCs细胞增殖抑制率达到77.5%,而相同浓度bFGF-SAP的抑制率仅46.1%;并且TGFα-SAP也比bFGF-SAP更明显抑制了培养SMCs的蛋白质合成。与空白对照组比较,3H-亮氨酸掺入量降低了47.3%;该浓度的TGFα-SAP和bF-GF-SAP对ECs却未显示出明显的细胞毒性作用。10-9mol/L的TGFα-SAP明显抑制了SMCs的DNA合成,使3H-TdR掺入量较对照组降低了29.5%,但同样浓度的bFGF-SAP仅使3H-TdR掺入量较空白对照组降低了9.86%。在10-7mol/L浓度下的TGFα-SAP未显示出明显对ECs的细胞毒性作用,而bFGF-SAP具有明显降低ECs的3H-TdR掺入量。结论:TGFα-SAP通过表皮生长因子受体介导具有明显增强的细胞毒性作用,其对增殖SMCs的细胞毒性作用较bFGF-SAP更为敏感,而在10-7mol/L浓度下,bFGF-SAP比TGFα-SAP对ECs显示出更明显的细胞毒性作用。本实验为TGFα-SAP在经皮冠脉成形术后再狭窄临床防治中的应用前景提供进一步实验依据。Objective：To compare the special cytotoxicity of TGFa-SAP with that of bFGF-SAP on proliferating vascular smooth muscle cells. Methods：Conjugation of saporin to TGFe was accomplished after derivatization of saporin, and TGFα with N-succinimidyl-3 （2-pyridyldithio） proprionate and final purification of the conjugate was achieved within Eppendorf Centrifugal Filter Tubes. Cytotoxicity assays were measured by MTS method. Effect of TGFα SAP and bFGF-SAP on protein synthesis of SMCs and ECs are measured by 3 H-leucine uptake. The influence of TGFα SAP and bFGF-SAP on DNA synthesis of SMCs and ECs was measured by 3 H-thymidine uptake. Results： Cytotoxicity assays testified TGFα- SAP could inhibit more remarkably proliferation of SMCs in culture, compared with bFGF-SAP. The values of ^3H-leucine incorporation of TGFα-SAP group （10^-9M） significantly decreased to 52.7% of the control group, suggesting that cellular protein synthesis obviously decreased as TGFα-SAP was added. But saporin did not affect cellular protein synthesis even at higher level （10^-3 M）. After SMCs were ex posed to TGFα-SAP and bFGF-SAP at the same level （10 ^-9 M）, the values of thymidine incorporation of TGFα-SAP group significantly decreased 29.5% compared with control group while that of bFGF-SAP group only decreased 9. 86% compared with control group. TGFα-SAP at higher level （107M） did not affect DNA synthesis of ECs compared with bFGF-SAP. Conclusion：The result indicated that TGFα- SAP possesses more sensitive and special cytotoxicity on proliferating SMCs than bFGF-SAP.

关 键 词：转化生长因子Α 皂草毒素蛋白 纤维母细胞生长因子 平滑肌细胞 再狭窄 

分 类 号：R541.4[医药卫生—心血管疾病]