L-EGCG对Lp(a)诱导大鼠系膜细胞增生及ICAM-1表达的影响

Effects of L-EGCG on proliferation glomerular mesangial cells of rat model induced by lipoprotein(a).

目的探讨儿茶素的主要成分L-表没食子基儿茶素没食子酸酯(L-EGCG)对脂蛋白(a)[Lp(a)]诱导的大鼠肾小球系膜细胞(GMCs)增生影响及可能机制。方法选定最佳剂量与最佳时间将GMCs分为对照组、Lp(a)组[Lp(a)(5.0μg/ml)]、L-EGCG组[Lp(a)(5.0μg/ml)+L-EGCG(200mg/L)处理]共三组分瓶培养。观察L-EGCG对GMCs增生率(MTT)、增生细胞核抗原(PCNA)阳性表达率及培养液上清的细胞间粘附因子-1(ICAM-1)浓度的影响,并与对照组相比较。结果在对照组、Lp(a)组及L-EGCG组,随着处理时间的延长,GMCs的MTT、PCNA、上清ICAM-1浓度呈增加趋势,经F检验,差异有统计学意义(P<0.01)。与对照组比较,应用Lp(a)5.0g/ml处理后,在不同的处理时间,Lp(a)组MTT计数、PCNA阳性率、ICAM-1均明显增高,经两两之间的q检验,差异有统计学意义(P<0.01)。与Lp(a)组比较,应用L-EGCG处理Lp(a)诱导的大鼠GMCs,在不同的处理时间,L-EGCG组MTT计数、PCNA阳性率、ICAM-1均明显降低,经两两之间的q检验,差异有统计学意义(P<0.01)。在Lp(a)组、L-EGCG组,培养上清ICAM-1浓度与PCNA阳性率及与反映系膜细胞增生指标的MTT呈正相关。结论Lp(a)能明显促进肾脏GMCs的增生,L-EGCG可明显抑制Lp(a)诱导的大鼠GMCs增生,此可能与影响ICAM-1表达有关。

Objective To investigate the effects of L- epigallocatechins gallic acid ester （L- EGCG ） on proliferation of glomerular mesangial cells （GMCs） of rat model induced by lipoprotein （a）[Lp（a）] and explore the possible mechanism of L- EGCG on the proliferation of rat GMCs by Lp（a）. Methods Rat GMCs were divided into three groups： control group, Lp （a） （5.0μg/ml） group, Lp（a） （5.0μg/ml） ＋ L - epigallocatechins gallic acid ester （ L - EGCG 200mg/L） group. After culture （ at the end of 12h, 24h, 48h, 60h and 72h）, collecting the cultured GMCs and suspension to observe the rate of GMCs proliferation（MTT）, the positive rate of proliferation cell nuclear antigen （PCNA） （immuno - histo - chemisty）, and to detect intercellular adhesion molecule-1（ICAM- 1） activity（ELISA）. Results At the end of experiment, MTT, the positive rate of PCNA and ICAM - 1 activity of GMCs were increased as culture time increased in three group. Compared with control group, MTT, positive rate of PCNA and ICAM - 1 activity of GMCs were more significantly enhanced in Lp（a） group. Compared with Lp（a） group, MTT, positive rate of PCNA and ICAM - 1 activity of GMCs were significantly decreased in Lp（a） ＋ L- EGCG group. ICAM - 1 activity showed positive correlationship with MTT and the positive rate of PCNA. Conclusion Lp（a） can significantly affect the rate of GMCs proliferation. L- EGCG can depress proliferation GMCs of rat model induced by Lp（a） and correct with ICAM- 1.