Tong Yang-Fei, Wang Yan, Ding Yuan-Yuan, Li Jing-Mei, Pan Xi-Chun, Lu Xiao-Lan, Chen Xiao-Hong, Liu Ya, Zhang Hai-Gang
Department of Pharmacology, College of Pharmacy, Third Military Medical University, Chongqing, China.
Department of Pharmacy, Chongqing Traditional Medicine Hospital, Chongqing, China.
Cell Physiol Biochem. 2017;42(4):1645-1656. doi: 10.1159/000479407. Epub 2017 Jul 24.
BACKGROUND/AIMS: Adult cardiomyocytes can re-enter cell cycle as stimulated by prohypertrophic factors although they withdraw from cell cycle soon after birth. p21WAF1/CIP1, a cyclin-dependent kinase inhibitor, has been implicated in cardiac hypertrophy, however, its precise contribution to this process remains largely unclear.
The gene expression profile in left ventricle (LV) of spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats was determined using quantitative PCR array and verified by real-time PCR and Western blotting. Hypertrophic response of H9c2 cells and neonatal rat ventricular myocytes (NRVM) were induced by angiotensin II (1 µmol/L). Cardiac hypertrophy of mice was elicited by isoproterenol (ISO) infusion (40 mg/kg per day for 14 days). p21-adenovirus and p21-siRNA were employed to transfect NRVM, and sterigmatocystin (STE, 3 mg/kg, ip, qd) was used to inhibit p21 activity. mRNA and protein expression levels of α- and β-myosin heavy chain (MHC), p21WAF1/CIP1, calcineurin (CaN) and atrial natriuretic peptide (ANP) were assayed by realtime PCR and WB, respectively.
Sixteen genes showed two-fold or greater changes between SHR and WKY rats, in which the expression of p21WAF1/CIP1 was upregulated by 4.15-fold (P=0.002) and reversed by losartan. Surface area, protein content, mRNA and protein expressions of β-MHC, ANP and p21WAF1/CIP1 in H9c2 cells treated with AngII elevated significantly compared with control group. p21-Ad transfection markedly increased the surface area and β-MHC mRNA expression of normal NRVMs, and p21-siRNA transfection decreased them in AngII-treated NRVMs. STE treatment decreased HW/BW and cross-sectional area, expression levels of β-MHC, ANP and p21 significantly in ISO-treated mice.
Our findings suggest that p21 facilitates the development of cardiac hypertrophy, and regulating the expression of p21 may be an approach to attenuate hypertrophic growth of cardiomyocytes.
背景/目的:成年心肌细胞在出生后不久就退出细胞周期,但在促肥大因子的刺激下可重新进入细胞周期。细胞周期蛋白依赖性激酶抑制剂p21WAF1/CIP1与心脏肥大有关,然而,其在这一过程中的具体作用仍不清楚。
使用定量PCR芯片测定自发性高血压大鼠(SHR)和Wistar-Kyoto(WKY)大鼠左心室(LV)的基因表达谱,并通过实时PCR和蛋白质印迹法进行验证。用血管紧张素II(1μmol/L)诱导H9c2细胞和新生大鼠心室肌细胞(NRVM)的肥大反应。通过输注异丙肾上腺素(ISO)(40mg/kg/天,共14天)诱发小鼠心脏肥大。用p21腺病毒和p21小干扰RNA转染NRVM,用杂色曲霉素(STE,3mg/kg,腹腔注射,每日一次)抑制p21活性。分别通过实时PCR和蛋白质印迹法检测α和β肌球蛋白重链(MHC)、p21WAF1/CIP1、钙调神经磷酸酶(CaN)和心钠素(ANP)的mRNA和蛋白质表达水平。
16个基因在SHR和WKY大鼠之间表现出两倍或更大的变化,其中p21WAF1/CIP1的表达上调了4.15倍(P=0.002),并被氯沙坦逆转。与对照组相比,用AngII处理的H9c2细胞中β-MHC、ANP和p21WAF1/CIP1的表面积、蛋白质含量、mRNA和蛋白质表达均显著升高。p21-Ad转染显著增加了正常NRVM的表面积和β-MHC mRNA表达,而p21小干扰RNA转染则降低了AngII处理的NRVM中的上述指标。STE处理显著降低了ISO处理小鼠的心脏重量/体重和横截面积、β-MHC、ANP和p21的表达水平。
我们的研究结果表明,p21促进心脏肥大的发展,调节p21的表达可能是减轻心肌细胞肥大生长的一种方法。