吲哚硫酸通过激活Caspase-8/GSDME通路诱导MC3T3-E1细胞焦亡并抑制成骨分化*

doi:10.3870/j.issn.1672-0741.25.07.002

华中科技大学学报（医学版） ›› 2026, Vol. 55 ›› Issue (4): 490-495.doi: 10.3870/j.issn.1672-0741.25.07.002

吲哚硫酸通过激活Caspase-8/GSDME通路诱导MC3T3-E1细胞焦亡并抑制成骨分化^*

王茜¹, 蔡纪平², 田晓晨^3△

¹河北医科大学实验中心,石家庄 050017
²石家庄医学高等专科学校中药学教研室,石家庄 050599
³石家庄市人民医院创伤急救科,石家庄 050031

收稿日期:2025-06-26 出版日期:2026-08-15 发布日期:2026-07-28
通讯作者: ^△E-mail:xin082323@163.com
作者简介:王茜,女,1986年生,医学硕士,实验师,E-mail:ighqi4006@163.com
基金资助:
^*河北省医学科学研究重点课题项目(No.20191466);河北省医学科学研究课题(No.20200865)

Indoxyl Sulfate Induces Pyroptosis and Inhibits Osteogenic Differentiation in MC3T3-E1 Cells via Activation of Caspase-8/GSDME Pathway

Wang Qian¹, Cai Jiping², Tian Xiaochen^3△

¹Experimental Center,Hebei Medical University,Shijiazhuang 050017,China
²Teaching and Research Office of Traditional Chinese Medicine, Shijiazhuang Medical College,Shijiazhuang 050599,China
³Department of Trauma and Emergency,Shijiazhuang People's Hospital,Shijiazhuang 050031,China

Received:2025-06-26 Online:2026-08-15 Published:2026-07-28
Contact: ^△E-mail:xin082323@163.com

摘要/Abstract

摘要： 目的探讨吲哚硫酸(IS)通过Caspase-8/GSDME通路诱导成骨前体细胞MC3T3-E1焦亡并抑制其成骨分化的机制。方法采用不同浓度(0、25、50、100 μmol/L)IS分别作用于MC3T3-E1细胞24、48、72 h,应用CCK-8法评估细胞增殖活力,Annexin Ⅴ-FITC/PI流式细胞术检测凋亡水平。根据预实验结果筛选最佳浓度及作用时间后,将细胞分为对照组、IS组及IS联合Caspase-8特异性抑制剂Z-IETD-FMK组。分别检测3组细胞的乳酸脱氢酶(LDH)释放水平、Caspase-8/GSDME相关蛋白表达水平,以及碱性磷酸酶(ALP)活性、成骨相关蛋白表达、钙化结节形成能力(ARS染色)等成骨功能指标。结果 IS在48 h作用时即表现出剂量依赖性的细胞毒性,100 μmol/L的抑制作用最强,显著降低细胞活力(P<0.01)并诱导凋亡(P<0.01)。Western blot显示IS上调Caspase-8、cleaved Caspase-8、消皮素E(Gasdermin E,GSDME)及其裂解产物GSDME-N表达(均P<0.01),同时促进LDH释放(P<0.01);Z-IETD-FMK干预后上述变化均被部分逆转(均P<0.05)。在成骨功能方面,IS显著抑制ALP活性(第3、5天,均P<0.01)、下调骨桥蛋白(OPN)、骨钙素(OCN)、骨唾液酸蛋白(BSP)的mRNA与蛋白表达水平(均P<0.01),并减少ARS染色钙化结节形成(P<0.01);阻断焦亡通路后成骨相关指标均有不同程度恢复(均P<0.05)。结论 IS通过激活Caspase-8/GSDME通路诱导MC3T3-E1细胞焦亡,造成细胞膜破裂与功能障碍,进而抑制其早期与晚期成骨分化能力。该通路可能为IS骨毒性效应的重要分子机制之一。

关键词: 吲哚硫酸, Caspase-8, GSDME, 细胞焦亡, 成骨分化, MC3T3-E1细胞

Abstract: Objective To investigate the mechanism by which indoxyl sulfate(IS)induces pyroptosis in MC3T3-E1 pre-osteoblasts and inhibits osteogenic differentiation through the Caspase-8/GSDME pathway.Methods MC3T3-E1 cells were treated with IS at various concentrations(0,25,50,and 100 μmol/L)for 24,48,and 72 h.Cell proliferative activity was assessed using the CCK-8 assay,and apoptosis was detected by Annexin Ⅴ-FITC/PI flowcytometry.After the optimal concentration and exposure time were selected based on preliminary experiments,the cells were divided into the control group,IS group,and IS combined with the Caspase-8-specific inhibitor Z-IETD-FMK group.Lactate dehydrogenase(LDH)release,Caspase-8/GSDME-related protein expression(Western blot),alkaline phosphatase(ALP)activity,osteogenesis-related protein expression,and calcium nodule formation(ARS staining)were detected in the three groups.Results IS showed dose-dependent cytotoxicity after 48 h of treatment,with the strongest inhibitory effect at 100 μmol/L,significantly reducing cell viability(P<0.01).Western blot analysis showed that IS upregulated the expression of Caspase-8,cleaved Caspase-8,gasdermin E(GSDME),and its cleaved product GSDME-N(all P<0.01),and promoted LDH release(P<0.01).These changes were partially reversed after Z-IETD-FMK intervention(all P<0.05).Regarding osteogenic function,IS significantly inhibited ALP activity on days 3 and 5(both P<0.01),downregulated the mRNA and protein expression levels of osteopontin(OPN),osteocalcin(OCN),and bone sialoprotein(BSP)(P<0.01),and reduced ARS-stained calcium nodule formation(P<0.01).After blockade of the pyroptosis pathway,osteogenesis-related indicators recovered to varying degrees(all P<0.05).Conclusion IS induces pyroptosis in MC3T3-E1 cells by activating the Caspase-8/GSDME pathway,causing cell membrane rupture and functional impairment,thereby inhibiting early and late osteogenic differentiation.This pathway may represent a key molecular mechanism underlying the osteotoxic effects of IS.

Key words: indoxyl sulfate, Caspase-8, GSDME, pyroptosis, osteogenic differentiation, MC3T3-E1 cells

中图分类号:

R681.1

王茜, 蔡纪平, 田晓晨. 吲哚硫酸通过激活Caspase-8/GSDME通路诱导MC3T3-E1细胞焦亡并抑制成骨分化^*[J]. 华中科技大学学报（医学版）, 2026, 55(4): 490-495.

Wang Qian, Cai Jiping, Tian Xiaochen. Indoxyl Sulfate Induces Pyroptosis and Inhibits Osteogenic Differentiation in MC3T3-E1 Cells via Activation of Caspase-8/GSDME Pathway[J]. Acta Medicinae Universitatis Scientiae et Technologiae Huazhong, 2026, 55(4): 490-495.

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吲哚硫酸通过激活Caspase-8/GSDME通路诱导MC3T3-E1细胞焦亡并抑制成骨分化^*

Indoxyl Sulfate Induces Pyroptosis and Inhibits Osteogenic Differentiation in MC3T3-E1 Cells via Activation of Caspase-8/GSDME Pathway

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