常染色体显性遗传性多囊肾病囊肿液对肾小管上皮细胞增殖和凋亡的影响

Effect of cystic fluid from patients with autosomal dominant polycystic kidney disease on proliferation and apoptosis of tubular cells cultured in vitro

目的 :观察常染色体显性遗传性多囊肾病 (ADPKD)囊肿液对肾小管细胞增殖、凋亡以及细胞周期的影响 ,以初步探讨 ADPKD囊肿发生、发展的机制。 方法 :用不同浓度的 ADPKD囊肿液处理 MDCK和 L L C- PK1 两株肾小管细胞 ,采用MTT法观察细胞增殖 ,用流式细胞术检测细胞周期和凋亡指数。 结果 :(1)与对照组相比 ,不同浓度 (10 %、2 0 %、4 0 %、6 0 % ,V/ V) ADPKD囊肿液作用 2 4 h能明显促进上述两株肾小管细胞增殖 ,并呈剂量依赖性 ;而且明显影响细胞周期 ,使 G0 ～ G1 期细胞增多、S期细胞减少。 (2 )高浓度 (40 %、6 0 % ) ADPKD囊肿液作用 4 8h对 MDCK细胞仍有上述作用 ,但对 L L C- PK1 细胞则无类似作用。 (3)不同浓度 ADPKD囊肿液均不诱导上述两株细胞凋亡。结论 :ADPKD囊肿液可剂量依赖性地促进肾小管细胞增殖 ,并在 2 4 h达作用高峰 ,但并不诱导肾小管细胞凋亡 ,其机制可能与囊肿液通过激活 G1 期细胞 ,使细胞未从 G1

Objective: To investigate the effect of cystic fluid from patients with autosomal dominant polycystic kidney disease(ADPKD) on proliferation, apoptosis and cell cycle of tubular cells cultured in vitro ,and to explore the mechanism of the cystogenesis and progression in ADPKD initially. Methods:Two tubular cell lines,MDCK and LLC PK 1, were incubated with various concentrations of cystic fluid from ADPKD patients. Cell proliferation was assessed by MTT colorimetric assay,cell cycle and apoptosis index were analyzed by flow cytometry. Results: Incubation with various concentrations of cystic fluid from ADPKD for 24 h promoted the proliferation of 2 tubular cell strains,regulated the cell cycle and provoked G 0 G 1 phase arrest. MDCK but not LLC PK 1 still had the same effect after incubation with higher concentrations of cystic fluid for 48 h. Various concentrations of cystic fluid could not induce apoptosis. Conclusion: Cystic fluid of ADPKD can stimulate the proliferation of tubular cells in a dose dependent manner and reach the peak effect after 24 h,but cannot lead to apoptosis,which might be related to regulation of cell cycle and activation of G 0 G 1 phase arrest.