Zhu Nan, Tang Yunhai, Yuan Weijie, Tang Zhihuan
Department of Nephrology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Nephrology, Jiading Branch of Shanghai General Hospital, Shanghai, China.
Ann Transl Med. 2022 Oct;10(19):1065. doi: 10.21037/atm-22-4352.
To investigate the effects and mechanism of high concentration glucose (HG), exogenous hydrogen peroxide (HO), and antioxidants on the cell growth (cell proliferation) of human peritoneal mesothelial cells (HPMCs).
All tests were conducted on cultured HPMCs (HMrSV5) . Various concentrations of glucose (0.1%, 1.35%, and 3.86%), HO (0.5 and 0.1 mmol/L), and antioxidants (pyruvate and catalase) were used in cell culture. Moreover, in order to study the interaction between these factors, HG and HO, HG and antioxidants, HG, HO, and antioxidants, were used respectively. After 12-24 h, phase-contrast microscopy was used to examine the morphological changes of HPMCs. DNA synthesis was detected by H-thymidine incorporation to measure cell proliferation, and flow cytometry was used to evaluate the proportion of cells in G phase. Furthermore, semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) was utilized to determine the mRNA expression of and [cyclin-dependent kinase inhibitors (CKIs)], while immunocytochemistry (ICC) and Western blotting were employed to measure the protein expression of p21 and p27.
HG or low-dose exogenous HO resulted in hypertrophy and senescence of HPMCs, resulting in similar morphological changes. Both HG and exogenous HO (0.5 mmol/L) inhibited the proliferation of HPMCs and led to G1 phase arrest of HPMCs. The proportion of cells in G phase increased. Moreover, HG enhanced the toxic effects of exogenous HO. Both HG and exogenous HO increased the expression of and . The addition of an antioxidant in HG medium arrested cells in the G phase and improved the inhibited cell proliferation.
Both HG and exogenous HO treatments can induce growth inhibition of HPMCs by arresting cell cycle progression, which is partially due to the increased expression of and . Thus, the effects of HG might be associated with endogenous reactive oxygen species (ROS), and it might be beneficial to use antioxidants in peritoneal dialysis (PD).
探讨高浓度葡萄糖（HG）、外源性过氧化氢（HO）及抗氧化剂对人腹膜间皮细胞（HPMCs）细胞生长（细胞增殖）的影响及其机制。
所有实验均在培养的HPMCs（HMrSV5）上进行。细胞培养中使用了不同浓度的葡萄糖（0.1%、1.35%和3.86%）、HO（0.5和0.1 mmol/L）以及抗氧化剂（丙酮酸和过氧化氢酶）。此外，为研究这些因素之间的相互作用，分别使用了HG与HO、HG与抗氧化剂、HG、HO与抗氧化剂。12 - 24小时后，用相差显微镜观察HPMCs的形态变化。通过³H - 胸腺嘧啶核苷掺入检测DNA合成以测量细胞增殖，并用流式细胞术评估G期细胞比例。此外，采用半定量逆转录聚合酶链反应（RT - PCR）测定细胞周期蛋白依赖性激酶抑制剂（CKIs）p21和p27的mRNA表达，同时采用免疫细胞化学（ICC）和蛋白质印迹法检测p21和p27的蛋白表达。
HG或低剂量外源性HO导致HPMCs肥大和衰老，产生相似的形态学变化。HG和外源性HO（0.5 mmol/L）均抑制HPMCs增殖并导致HPMCs G1期阻滞。G期细胞比例增加。此外，HG增强了外源性HO的毒性作用。HG和外源性HO均增加了p21和p27的表达。在HG培养基中添加抗氧化剂使细胞停滞于G期并改善了受抑制的细胞增殖。
HG和外源性HO处理均可通过阻滞细胞周期进程诱导HPMCs生长抑制，这部分归因于p21和p27表达增加。因此，HG的作用可能与内源性活性氧（ROS）有关，在腹膜透析（PD）中使用抗氧化剂可能有益。
