目的探讨表没食子儿茶素没食子酸脂(EGCG)对高糖培养的系膜细胞(GMCs)增殖及周期素激酶抑制剂p27蛋白表达的作用。方法以大鼠肾系膜细胞为实验对象,将GMCs分成6组,分别刺激48βh后,用四甲基偶氮唑蓝(MTT)测定GMCs增殖情况,Western blot法测定p27蛋白表达。结果作用48βh后,高糖明显诱导GMCs增殖并上调p27蛋白表达,不同浓度的EGCG能抑制高糖诱导的细胞增生,并抑制p27蛋白表达。结论EGCG能抑制高糖诱导的GMCs增殖,并下调p27蛋白表达,提示EGCG可通过抑制p27表达而减少细胞外基质的积聚,延缓糖尿病肾小球肥大和肾小球硬化。
The effect of epigallocatechin-3-gallate (EGCG) on rat mesangial cells (GMCs) proliferation cultured in high glucose condition and the expression of cyclin kinase inhibitor p27(p27) protein were investigated. The GMCs were divided into 6 groups. 48 hours later, the proliferation activity of GMCs was observed by MTT assay, the expression of p27 protein was enhanced by Western blot. Results showed that compared with normal glucose group, high glucose treatment significantly increased GMCs proliferation activity and the expression of p27 protein. EGCG in different concentration suppressed the proliferation of GMCs and down-regulated the expression of p27. It can be concluded that EGCG can suppress the proliferation activity of GMCs cultured under high glucose and decrease the expression of p27 protein, which implied EGCG probably decrease the accumulation of extracellular matrix (ECM) and slowed the progession of progression of glomerular hypertrophy and glomerulosclerosis in diabetic nephropathy.
[1] Awazu M, Omori S, Ishikura K, et al. The lack of cyclin kinase inhibitor p27(Kip1) ameliorates progression of diabetic nephropathy[J]. J Am Soc Nephrol. 2003, 14(3): 699~708.
[2] Shankland JS.Cell-cycle control and renal disease[J]. Kidney Int, 1997, 52(2): 294~308
[3] Wolf G, Reinking R, Zahner G, et al. Erk 1,2 phosphorylates p27(Kip1): Functional evidence for a role in high glucose-induced hypertrophy of mesangial cells[J]. Diabetologia. 2003, 46(8): 1090~1099.
[4] Wolf G, Schroeder R, Ziyadeh FN, et al. High glucose stimulates expression of p27Kip1 in cultured mouse mesangial cells: relationship to hypertrophy[J]. Am J Physiol. 1997, 273(3 Pt 2): F348~56.
[5] Wolf G, Schroeder R, Zahner G, et al. High glucose-induced hypertrophy of mesangial cells requires p27(Kip1), an inhibitor of cyclin-dependent kinases[J]. American Journal of Pathology. 2001, 158(3): 1091~1100.