摘要
目的 探讨组蛋白甲基化酶同源序列增强子2(enhancer of zeste homolog 2,EZH2)通过调控细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)/低氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)信号通路在腹膜透析相关腹膜纤维化中的作用。 方法 细胞实验：人腹膜间皮细胞分为对照组、转化生长因子β(transforming growth factor-β,TGF-β)组及靶向EZH2的小干扰RNA(short interfering RNAtargeting EZH2,siEZH2)组。动物实验：小鼠腹膜纤维化模型分为对照组、模型组及模型+EZH2敲降组。qPCR和Western Blot检测EZH2、ERK、HIF-1α、α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、基质金属蛋白酶2(matrix metallo proteinase,MMP2)、白细胞介素-6(interleukin-6,IL-6)及单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)的表达；HE与Masson染色观察腹膜病变。 结果 细胞实验：TGF-β诱导细胞向成纤维细胞转化，siEZH2干预后减弱。与对照组比较，TGF-β组EZH2(mRNA:t=4.473,P=0.001； 蛋白:t=22.39,P＜0.001)、HIF-1α(t=5.873,P=0.004；t=3.763,P=0.019)、α-SMA(t=20.71,P＜0.001；t=4.587,P=0.010)、MMP2(t=47.500,P＜0.001；t=3.994,P=0.016)及p-ERK(蛋白:t=3.355,P=0.028)均升高，IL-6、MCP-1亦上调(均P＜0.05)。与TGF-β组比较，TGF-β+siEZH2组上述指标均降低：EZH2(mRNA:t=32.430,P＜0.001； 蛋白:t=4.886,P=0.008)、HIF-1α(t=6.732,P=0.003；t=0.019,P=0.001)、α-SMA(t=25.760,P＜0.001；t=2.809,P=0.048)、MMP2(t=45.30,P＜0.001；t=3.119,P=0.035)、p-ERK(蛋白:t=2.851,P=0.046)，IL-6、MCP-1亦下调(均P＜0.05)。动物实验：模型组腹膜间皮层增厚、纤维化及炎症浸润，EZH2敲降组改善。与对照组比较，模型组EZH2(mRNA:t=12.340,P＜0.001；蛋白:t=93.560,P＜0.001)、HIF-1α(t=3.891,P=0.017；t=35.250,P＜0.001)、α-SMA(t=17.540,P＜0.001；t=15.020,P＜0.001)、MMP2(t=47.330,P＜0.001；t=61.120,P＜0.001)及p-ERK蛋白(t=102.800,P＜0.001)均升高。与模型组比较，EZH2敲降组上述指标均降低：EZH2(mRNA:t=3.313,P=0.029； 蛋白:t=29.040,P＜0.001)、HIF-1α(t=3.928,P=0.017；t=38.520,P＜0.001)、α-SMA(t=6.617,P=0.002；t=5.570,P=0.005)、MMP2(t=5.406,P=0.005；t=10.150,P＜0.001)、p-ERK(蛋白:t=59.460,P＜0.001)。ERK mRNA无组间差异(P＞0.05)。 结论 EZH2通过激活ERK/HIF-1α通路促进腹膜纤维化，上调HIF-1α、α-SMA、MMP2及炎症因子；敲降EZH2可抑制该通路减轻纤维化。靶向EZH2/ERK/HIF-1α通路或为治疗新策略。
Abstract
Objective To investigate the role of enhancer of zeste homolog 2 (EZH2) through regulating the extracellular signal-regulated kinase (ERK)/hypoxia inducible factor-1α (HIF-1α) signaling pathway in the pathogenesis of peritoneal dialysis-associated peritoneal fibrosis. Methods Cell studies: human peritoneal mesothelial cells were divided into 3 groups, control group, TGF-β group, and TGF-β+siEZH2 (TGF-β+small interference RNA targeting EZH2) group. Animal studies: the mouse peritoneal fibrosis model was divided into 3 groups, control group, model group, and model+EZH2 knockdown group. The expressions of EZH2, ERK, HIF-1α, α-SMA (α-smooth muscle actin), MMP2 (matrix metallo proteinase 2), IL-6 (interleukin-6), and MCP-1 (monocyte chemoattractant protein-1) were detected by qPCR and western blot. Peritoneal tissues were stained by HE and Masson method. Results Cell studies: TGF-β induced the transformation of human peritoneal mesothelial cells to fibroblasts, which was attenuated by knockdown of EZH2. Compared with those in the control group, the TGF-β group showed the increase of mRNAs and proteins of EZH2 (t=4.473 and 22.39, P=0.001 and ＜0.001), HIF-1α (t=5.873 and 3.763, P=0.004 and 0.019), α-SMA (t=20.71 and 4.587, P＜0.001 and =0.010), and MMP2 (t=47.500 and 3.994, P＜0.001 and =0.016), as well as the increase of p-ERK (t=3.355, P=0.028)； IL-6 and MCP-1 were also upregulated (P＜0.05). Compared with those in the TGF-β group, the TGF-β+siEZH2 group exhibited the reduction of mRNAs and proteins of EZH2 (t=32.430 and 4.886, P＜0.001 and=0.008), HIF-1α (t=6.732 and 0.019, P=0.003 and 0.001), α-SMA (t=25.760 and 2.809, P＜0.001 and =0.048), and MMP2 (t=45.30 and 3.119, P＜0.001 and =0.035), as well as the reduction of p-ERK (t=2.851, P=0.046)； IL-6 and MCP-1 were also downregulated (P＜0.05). No intergroup differences were observed in ERK mRNA (P＞0.05). Animal studies: the model group exhibited thickening of peritoneal mesothelial layer, fibrosis and inflammatory infiltration, which were ameliorated in the model+EZH2 knockdown group. Compared with the control group, the model group showed the increase of mRNAs and proteins of EZH2 (t=12.340 and 93.560, P＜0.001), HIF-1α (t=3.891 and 35.250, P=0.017 and ＜0.001), α-SMA (t=17.540 and 15.020, P＜0.001), and MMP2 (t=47.330 and 61.120, P＜0.001), as well as the increase of p-ERK (t=102.800, P＜0.001). Compared with the model group, the model+EZH2 knockdown group exhibited the reduction of mRNAs and proteins of EZH2 (t=3.313 and 29.040, P=0.029 and ＜0.001), HIF-1α (t=3.928 and 38.520, P=0.017 and ＜0.001), α-SMA (t=6.617 and 5.570, P=0.002 and 0.005), and MMP2 (t=5.406 and 10.150, P=0.005 and ＜0.001), as well as the reduction of p-ERK (t=59.460, P＜0.001). No intergroup differences were observed in ERK mRNA (P＞0.05). Conclusion EZH2 promotes peritoneal fibrosis by activating the ERK/HIF-1α pathway to upregulate the expressions of HIF-1α, α-SMA, MMP2 and inflammatory factors. Knockdown of EZH2 inhibits this pathway and attenuates fibrosis. Targeting the EZH2/ERK/HIF-1α pathway may represent a novel therapeutic strategy.
关键词
组蛋白甲基化酶同源序列增强子2 /
细胞外信号调节激酶 /
低氧诱导因子-1α /
腹膜透析 /
腹膜纤维化
Key words
EZH2 /
ERK /
HIF-1α /
Peritoneal dialysis /
Peritoneal fibrosis
傅碧玲 谢婷妃 许云鹏 张燕子 陈继红.
组蛋白甲基化酶同源序列增强子2介导调控细胞外信号调节激酶/低氧诱导因子-1α信号通路促进腹膜透析相关腹膜纤维化的机制研究[J]. 中国血液净化. 2026, 25(06): 508-513 https://doi.org/10.3969/j.issn.1671-4091.2026.06.013
FU Bi-ling, XIE Ting-fei, XU Yun-peng, ZHANG Yan-zi, CHEN Ji-hong.
Mechanism of EZH2-mediated ERK/HIF-1α signaling pathway in promoting peritoneal dialysis-associated peritoneal fibrosis[J]. Chinese Journal of Blood Purification. 2026, 25(06): 508-513 https://doi.org/10.3969/j.issn.1671-4091.2026.06.013
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基金
创建广东省基层医疗卫生机构“慢性肾脏病防治示范区-智慧腹膜透析中心的建设与管理”项目(440306241171588000012)； 深圳市宝安区医学会研究项目(BAYXH2023013)
