原发性醛固酮增多症患者浦肯野细胞蛋白4表达与细胞凋亡关系研究

A study of the correlation between Purkinji cell protein 4 and cell apoptosis in primary aldosteronism

目的探讨原发性醛固酮增多症(primary aldosteronism,PA)的两种主要亚型——醛固酮瘤(aldosterone-producing adenoma,APA)及特发性醛固酮增多症(idiopathic hyperaldosteronism,IHA)患者组织中浦肯野细胞蛋白(Purkinje cell protein,PCP)4的表达水平,并研究其与肾上腺细胞凋亡的关系。方法检测10例正常肾上腺、7例IHA、16例APA患者的组织样本中PCP4的信使RNA(messenger RNA,mRNA)水平。将两种PCP4特异性小干扰RNA(small interfering RNA,siRNA)及一种非特异性siRNA,分别转染人肾上腺皮质小细胞癌细胞系SW-13及人肾上腺皮质腺癌细胞系NCI-H295R,检测PCP4的mRNA及蛋白水平,检测各组转染后的凋亡细胞数量。结果 PCP4在APA和IHA中的表达高于正常肾上腺组织。RNA干扰在体外可有效抑制PCP4表达。PCP4可抑制NCI-H295R、SW-13细胞凋亡。结论 APA、IHA患者较正常者肾上腺组织中PCP4的表达高,PCP4可能在PA发病机制中起着重要作用,有望成为新的治疗靶点。

Objective To investigate the expression level of Purkinje cell protein(PCP) 4 in the tissue of patients with two main subtypes of primary aldosteronism(PA),aldosterone-producing adenoma(APA) and idiopathic hyperaldosteronism(IHA),and to study its relationship with adrenal cell apoptosis.Methods The levels of messenger RNA(mRNA) of PCP4 were detected in tissue samples of 10 normal adrenal glands,7 patients with IHA and 16 patients with APA.Two kinds of PCP4 specific small interfering RNA(siRNA) and one non-specific siRNA were transfected into human adrenocortical small cell carcinoma cell line SW-13 and human adrenocortical adenocarcinoma cell line NCI-H295R respectively.The mRNA and protein levels of PCP4 were detected,and the number of apoptotic cells was detected after transfection.Results The expression of PCP4 in APA and IHA was higher than that in normal adrenal tissue.RNA interference can effectively inhibit the expression of PCP4 in vitro.PCP4 can inhibit the apoptosis of NCI-H295R and SW-13 cells.Conclusion The expression of PCP4 in adrenal tissue of patients with APA and IHA is higher than that of normal subjects.PCP4 may play an important role in the pathogenesis of PA and is expected to become a new therapeutic target.