摘要
目的基质金属蛋白酶-2（matrix metalloprote inase-2,MMP-2）及金属蛋白酶组织抑制剂-1（tissue inh ib itor of m et-alloprote inase-1,TIMP-1）失衡可导致细胞外基质沉积,是组织纤维化包括腹膜组织纤维化发生发展的重要机制之一。通过体外细胞培养,观察丹参酮ⅡA对人腹膜间皮细胞增殖活性及MMP-2、TIMP-1表达的影响,并通过在腹膜透析液（peritoneal dial-ysis solution,PDS）中添加丹参酮ⅡA了解其对腹膜透析（peritoneal dialysis,PD）患者腹腔MMP-2和TIMP-1的影响。方法取6例择期手术患者的网膜体外培养腹膜间皮细胞,第3代细胞用于实验,细胞同步后分成对照组、PDS组、丹参酮ⅡA 1mg/L组（丹参酮ⅡA终浓度为1mg/L）、丹参酮ⅡA 5mg/L组（丹参酮ⅡA终浓度为5mg/L）、丹参酮ⅡA 10 mg/L组（丹参酮ⅡA终浓度为10 mg/L）5组（每组6例）进行干预。ELISA法检测各组上清液中MMP-2、TIMP-1的含量,RT-PCR检测各组细胞MMP-2、TIMP-1mRNA的表达。前瞻性观察38例持续非卧床PD患者,随机分为实验组和对照组,每组19例,进行为期6周的研究。实验组在前3周使用添加丹参酮ⅡA注射液（10mg/2L）的PDS治疗,4~6周使用常规PDS治疗,对照组均不加药治疗6周,分别在开始时、第3周末和第6周末检测2组患者透出液MMP-2、TIMP-1的浓度,并进行比较。结果与对照组比较,PDS组人腹膜间皮细胞产生MMP-2明显增加（P〈0.05）,MMP-2/TIMP-1比值明显升高（P〈0.05）,表达MMP-2 mRNA明显升高（P〈0.05）;添加丹参酮ⅡA干预后,丹参酮ⅡA 1 mg/L组、5 mg/L组、10 mg/L组MMP-2产生与PDS组比较有显著下降（P〈0.05）,MMP-2/TIMP-1比值明显下降（P〈0.05）,MMP-2mRNA表达水平显著下调（P〈0.01）,TIMP-1mRNA表达水平显著上调（P〈0.05）。用药后实验组患者透出液MMP-2较用药前有明显降低（P〈0.05）,透出液TIMP-1无明显变化,停药3周后,透出液MMP-2较用药3周后明显升高（P〈0.05）。结论丹参酮ⅡA可影响人腹膜间皮细胞MMP-2、TIMP-1mRNA的表达,调节MMP-2/TIMP-1平衡,改善人腹膜间皮细胞的增殖活性,显著降低PD患者腹腔内高MMP-2状态。
Objective The imbalance of matrix metalloproteinases-2（MMP-2） and its tissue inhibitor-1（TIMP-1） is responsible for the deposition of extracellular matrix components involved in tissue fibrosis,including peritoneal fibrosis.This study was to investigate the effects of Tanshinone ⅡA on cell proliferation activity and the secretion of MMP-2 and TIMP-1 in human peritoneal mesothelial cells（HPMC） cultured in vitro with peritoneal dialysate（PDS） as well as the impact of Tanshinone ⅡA on MMP-2 and TIMP-1 of peritoneal cavity in continuous ambulatory peritoneal dialysis（CAPD） patients.Methods HPMCs were cultured in vitro and divided into five groups： control,PDS,and 1mg/L,5mg/L and 10mg/L Tanshinone ⅡA.After 48 h incubation,the levels of MMP-2 and TIMP-1 in the supernatant and the expressions of MMP-2 and TIMP-1mRNA in the HPMCs were determined by ELISA and RT-PCR,respectively.Thirty-eight stable CAPD patients were randomized to an experimental and a control group of equal number,the former treated by injection of PDS with Tanshinone ⅡA（10 mg/2L） for 3 weeks and PDS alone for another 3,while the latter without Tanshinone ⅡA throughout the 6 weeks.The concentrations of MMP-2 and TIMP-1 in the effluent were assayed at 0,3 and 6 weeks and compared among different groups.Results Significant increases were observed in the level of MMP-2 in the supernatant,the ratio of MMP-2 to TIMP-1,and the expression of MMP-2mRNA in HPMCs in the PDS group as compared with the control（P0.05）.After Tanshinone ⅡA intervention,the concentration of MMP-2 in the effluent was significantly lower than that before treatment（P0.05） and higher than that at 6 weeks（P0.05）,while no significant difference was found in TIMP-1 in the effluent.Conclusion Tanshinone ⅡA can significantly decrease the PDS-induced high expression of MMP-2mRNA and improve the proliferation activity of HPMCs,adjust the balance of MMP-2 and TIMP-1,and decrease the level of MMP-2 in peritoneal cavity in CAPD patients.
出处
《医学研究生学报》
CAS
2011年第9期913-918,共6页
Journal of Medical Postgraduates
基金
南京市医学科技发展重点项目(ZKX08026)
