Article(id=1291753196872028663, tenantId=1146029695717560320, journalId=1291416913693474840, issueId=1291753151179280707, articleNumber=PA20260408_3CpQMjfK, orderNo=null, doi=10.3969/j.issn.1671-7856.2026.06.007, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1754323200000, receivedDateStr=2025-08-05, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1785907561748, onlineDateStr=2026-08-05, pubDate=1774800000000, pubDateStr=2026-03-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1785907561747, onlineIssueDateStr=2026-08-05, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1785907561747, creator=admin, updateTime=1785907561747, updator=admin, issue=Issue{id=1291753151179280707, tenantId=1146029695717560320, journalId=1291416913693474840, year='2026', volume='36', issue='6', pageStart='1', pageEnd='144', issueExtLink='null', onlineDate='null', pubDate='1774800000000', pubDateStr='2026-03-30', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1785907550854, creator='ztmeta', updateTime=1786331902788, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1293533012289941791, tenantId=1146029695717560320, journalId=1291416913693474840, issueId=1291753151179280707, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1293533012289941792, tenantId=1146029695717560320, journalId=1291416913693474840, issueId=1291753151179280707, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=69, endPage=81, ext={EN=ArticleExt(id=1291753206246298106, articleId=1291753196872028663, tenantId=1146029695717560320, journalId=1291416913693474840, language=EN, title=Effect of free Heme on sepsis-induced immunosuppression model mice based on regulation of macrophage PANoptosis by NLRP12, columnId=1292760394137887571, journalTitle=Chinese Journal of Comparative Medicine, columnName=Reseaech Reports, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the effect of free Heme on sepsis-induced immunosuppression model mice based on regulation of macrophage PANoptosis by NLR family pyrin domain containing 12 (NLRP12). Methods A sepsis mouse model was constructed and the mice were divided into Model, Heme, Heme+empty plasmid (Heme+si-NC), and Heme+NLRP12 low-expression plasmid groups (Heme+si-NLRP12) (n=12 mice per group). Another 12 mice with exposed cecum without ligation or puncture were considered as the sham surgery group (Sham). Serum levels of inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). The CD4+/CD8+ ratio in the blood was detected by flow cytometry, pathological changes in lung tissue were detected by hematoxylin/eosin staining, apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining, and expression levels of PANoptosis-related proteins in lung tissue were detected by Western blot. To clarify the mechanism by which heme regulates NLRP12, sepsis cell models were induced using lipopolysaccharide (LPS)-infected mouse bone marrow-derived macrophages (BMDM). Model mice were divided into LPS, LPS+Heme, LPS+Heme+si-NC group, and LPS+Heme+si-NLRP12 groups, with mice administered normally cultivated BMDMs as the Control group. Levels of inflammatory factors in the BMDM supernatant were detected by ELISA, cell viability was detected by Cell Counting Kit-8 assay, apoptosis rate was detected by TUNEL staining, and expression levels of PANoptosis-related proteins were detected by Western blot.

Results

Compared with the Model group, rats in the heme group showed increased serum levels of inflammatory factors, aggravated lung tissue injury and cell apoptosis, and progression of cell PANoptosis, while the CD4+/CD8+ ratio was reduced (P<0.05). Compared with the Heme+si-NC group, pathological lung tissue damage and the process of PANoptosis were alleviated in the Heme+si-NLRP12 group, serum levels of inflammatory factors, and the apoptosis rate of lung tissue cells were decreased, while the CD4+/CD8+ ratio was increased (P<0.05). Cell experiments showed that the cell survival rate was significantly decreased in the LPS group, while the process of cell PANoptosis and levels of inflammatory factors in the supernatant were increased (P<0.05). Compared with the LPS group, the apoptosis rate, process of PANoptosis, and levels of inflammatory factors were further increased in the LPS+Heme group (P<0.05). Compared to the LPS+Heme+si-NC group, the LPS + Heme + si-NLRP12 group exhibited significantly increased cell viability, reduced apoptosis and supernatant inflammatory factor levels, along with decreased expression of NLRP12, Bax, Caspase-8, p-RIPK3, and GSDMD-N(P<0.05).

Conclusions

Heme may induce macrophage PANoptosis by upregulating NLRP12, thereby enhancing immune suppression in sepsis.

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目的

基于含NLR家族pyrin结构域12(NLRP12)调控巨噬细胞泛凋亡探讨游离血红素(Heme)对脓毒症免疫抑制模型小鼠的影响。

方法

构建脓毒症模型小鼠并将其分为模型(Model)组、游离血红素(Heme)组、游离血红素+阴性对照质粒转染(Heme + si-NC)组、游离血红素+ NLRP12低表达质粒转染(Heme+si-NLRP12)组,每组12只小鼠。另取12只仅暴露盲肠不进行结扎和穿刺的小鼠作为假手术(Sham)组。检测各组小鼠血清中炎症因子水平(ELISA法)、CD4+/CD8+比值(流式细胞术法)、肺组织病理变化(HE染色法)、细胞凋亡情况(TUNEL染色法)、肺组织中泛凋亡相关蛋白表达量(Western blot法)。为进一步明确Heme调节NLRP12的作用机制,以LPS感染小鼠骨髓来源巨噬细胞(BMDM)诱导脓毒症细胞模型,并将其分为LPS组、LPS+Heme组、LPS + Heme + si-NC组、LPS + Heme + si-NLRP12组,以正常培养的BMDM作为对照(Control)组。检测BMDM上清炎性因子水平(ELISA法)、细胞存活率(CCK-8法)、凋亡率(TUNEL法)及泛凋亡相关蛋白表达水平(Western blot法)。

结果

与Model组比较,Heme组可进一步提高脓毒症小鼠血清炎症因子水平、加重肺组织损伤和细胞凋亡、促进细胞的泛凋亡进展,降低血清CD4+/CD8+比值(P< 0.05);与Heme+si-NC组比较,Heme+si-NLRP12组脓毒症小鼠肺组织病理损伤及泛凋亡过程减轻、血清炎症因子水平及肺组织细胞凋亡率下降、血清CD4+/CD8+比值升高(P<0.05)。细胞实验显示LPS组细胞存活率显著下降、细胞泛凋亡过程及上清炎性因子水平明显增加(P<0.05);LPS+Heme组相较于LPS组细胞凋亡率、泛凋亡过程及炎性因子水平进一步增加(P<0.05);与LPS+Heme+si-NC组比较,LPS+Heme+si-NLRP12组细胞存活率显著增加、凋亡率及上清炎性因子水平降低,NLRP12、Bax、Caspase-8、p-RIPK3及GSDMD-N表达量显著下降(P< 0.05)。结论Heme可能通过上调NLRP12的表达诱导巨噬细胞泛凋亡,进而加强脓毒症的免疫抑制作用。

, authors=易呈志1, 李鸣1, *, 吴威2, 刘鹏1, authorsList=易呈志, 李鸣, 吴威, 刘鹏, authorCompany=null, correspAuthors=李鸣, authorNote=

易呈志(1982—),男,硕士,主治医师,研究方向:创伤急危重症,脓毒症。E-mail:

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李鸣(1970—),男,本科,主任医师,研究方向:肝胆、胃肠等及疾病的诊治及腹腔镜治疗。E-mail:
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基于NLRP12调控巨噬细胞泛凋亡探讨游离血红素对脓毒症免疫抑制模型小鼠的影响
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易呈志 1 , 李鸣 1, * , 吴威 2 , 刘鹏 1
中国比较医学杂志 | 研究报告 2026,36(6): 69-81
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中国比较医学杂志 |研究报告 2026 , 36 (6) : 69 -81
基于NLRP12调控巨噬细胞泛凋亡探讨游离血红素对脓毒症免疫抑制模型小鼠的影响
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易呈志1 , 李鸣1, * , 吴威2, 刘鹏1
作者信息
  • 1.武汉市第四医院古田院区急诊外科,武汉 430030
  • 2.武汉市第四医院古田院区急诊内科,武汉 430030
通讯作者:
李鸣(1970—),男,本科,主任医师,研究方向:肝胆、胃肠等及疾病的诊治及腹腔镜治疗。E-mail:
作者简介:

易呈志(1982—),男,硕士,主治医师,研究方向:创伤急危重症,脓毒症。E-mail:

Effect of free Heme on sepsis-induced immunosuppression model mice based on regulation of macrophage PANoptosis by NLRP12
Chengzhi YI1 , Ming LI1, * , Wei WU2, Peng LIU1
Affiliations
  • 1.Emergency Surgery, Wuhan Fourth Hospital Gutian Campus, Wuhan 430030, China
  • 2.Emergency Internal Medicine, Wuhan Fourth Hospital Gutian Campus, Wuhan 430030
出版时间: 2026-03-30 doi: 10.3969/j.issn.1671-7856.2026.06.007
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目的

基于含NLR家族pyrin结构域12(NLRP12)调控巨噬细胞泛凋亡探讨游离血红素(Heme)对脓毒症免疫抑制模型小鼠的影响。

方法

构建脓毒症模型小鼠并将其分为模型(Model)组、游离血红素(Heme)组、游离血红素+阴性对照质粒转染(Heme + si-NC)组、游离血红素+ NLRP12低表达质粒转染(Heme+si-NLRP12)组,每组12只小鼠。另取12只仅暴露盲肠不进行结扎和穿刺的小鼠作为假手术(Sham)组。检测各组小鼠血清中炎症因子水平(ELISA法)、CD4+/CD8+比值(流式细胞术法)、肺组织病理变化(HE染色法)、细胞凋亡情况(TUNEL染色法)、肺组织中泛凋亡相关蛋白表达量(Western blot法)。为进一步明确Heme调节NLRP12的作用机制,以LPS感染小鼠骨髓来源巨噬细胞(BMDM)诱导脓毒症细胞模型,并将其分为LPS组、LPS+Heme组、LPS + Heme + si-NC组、LPS + Heme + si-NLRP12组,以正常培养的BMDM作为对照(Control)组。检测BMDM上清炎性因子水平(ELISA法)、细胞存活率(CCK-8法)、凋亡率(TUNEL法)及泛凋亡相关蛋白表达水平(Western blot法)。

结果

与Model组比较,Heme组可进一步提高脓毒症小鼠血清炎症因子水平、加重肺组织损伤和细胞凋亡、促进细胞的泛凋亡进展,降低血清CD4+/CD8+比值(P< 0.05);与Heme+si-NC组比较,Heme+si-NLRP12组脓毒症小鼠肺组织病理损伤及泛凋亡过程减轻、血清炎症因子水平及肺组织细胞凋亡率下降、血清CD4+/CD8+比值升高(P<0.05)。细胞实验显示LPS组细胞存活率显著下降、细胞泛凋亡过程及上清炎性因子水平明显增加(P<0.05);LPS+Heme组相较于LPS组细胞凋亡率、泛凋亡过程及炎性因子水平进一步增加(P<0.05);与LPS+Heme+si-NC组比较,LPS+Heme+si-NLRP12组细胞存活率显著增加、凋亡率及上清炎性因子水平降低,NLRP12、Bax、Caspase-8、p-RIPK3及GSDMD-N表达量显著下降(P< 0.05)。结论Heme可能通过上调NLRP12的表达诱导巨噬细胞泛凋亡,进而加强脓毒症的免疫抑制作用。

血红素  /  脓毒症免疫抑制小鼠  /  巨噬细胞泛凋亡  /  含NLR家族pyrin结构域12
Objective

To investigate the effect of free Heme on sepsis-induced immunosuppression model mice based on regulation of macrophage PANoptosis by NLR family pyrin domain containing 12 (NLRP12). Methods A sepsis mouse model was constructed and the mice were divided into Model, Heme, Heme+empty plasmid (Heme+si-NC), and Heme+NLRP12 low-expression plasmid groups (Heme+si-NLRP12) (n=12 mice per group). Another 12 mice with exposed cecum without ligation or puncture were considered as the sham surgery group (Sham). Serum levels of inflammatory factors were detected by enzyme-linked immunosorbent assay (ELISA). The CD4+/CD8+ ratio in the blood was detected by flow cytometry, pathological changes in lung tissue were detected by hematoxylin/eosin staining, apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining, and expression levels of PANoptosis-related proteins in lung tissue were detected by Western blot. To clarify the mechanism by which heme regulates NLRP12, sepsis cell models were induced using lipopolysaccharide (LPS)-infected mouse bone marrow-derived macrophages (BMDM). Model mice were divided into LPS, LPS+Heme, LPS+Heme+si-NC group, and LPS+Heme+si-NLRP12 groups, with mice administered normally cultivated BMDMs as the Control group. Levels of inflammatory factors in the BMDM supernatant were detected by ELISA, cell viability was detected by Cell Counting Kit-8 assay, apoptosis rate was detected by TUNEL staining, and expression levels of PANoptosis-related proteins were detected by Western blot.

Results

Compared with the Model group, rats in the heme group showed increased serum levels of inflammatory factors, aggravated lung tissue injury and cell apoptosis, and progression of cell PANoptosis, while the CD4+/CD8+ ratio was reduced (P<0.05). Compared with the Heme+si-NC group, pathological lung tissue damage and the process of PANoptosis were alleviated in the Heme+si-NLRP12 group, serum levels of inflammatory factors, and the apoptosis rate of lung tissue cells were decreased, while the CD4+/CD8+ ratio was increased (P<0.05). Cell experiments showed that the cell survival rate was significantly decreased in the LPS group, while the process of cell PANoptosis and levels of inflammatory factors in the supernatant were increased (P<0.05). Compared with the LPS group, the apoptosis rate, process of PANoptosis, and levels of inflammatory factors were further increased in the LPS+Heme group (P<0.05). Compared to the LPS+Heme+si-NC group, the LPS + Heme + si-NLRP12 group exhibited significantly increased cell viability, reduced apoptosis and supernatant inflammatory factor levels, along with decreased expression of NLRP12, Bax, Caspase-8, p-RIPK3, and GSDMD-N(P<0.05).

Conclusions

Heme may induce macrophage PANoptosis by upregulating NLRP12, thereby enhancing immune suppression in sepsis.

Heme  /  sepsis immunosuppressed mice  /  macrophage PANoptosis  /  NLR family pyrin domain containing 12
易呈志, 李鸣, 吴威, 刘鹏. 基于NLRP12调控巨噬细胞泛凋亡探讨游离血红素对脓毒症免疫抑制模型小鼠的影响. 中国比较医学杂志, 2026 , 36 (6) : 69 -81 . DOI: 10.3969/j.issn.1671-7856.2026.06.007
Chengzhi YI, Ming LI, Wei WU, Peng LIU. Effect of free Heme on sepsis-induced immunosuppression model mice based on regulation of macrophage PANoptosis by NLRP12[J]. Chinese Journal of Comparative Medicine, 2026 , 36 (6) : 69 -81 . DOI: 10.3969/j.issn.1671-7856.2026.06.007
  • 武汉市医学科学研究项目(WX23Z53)
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doi: 10.3969/j.issn.1671-7856.2026.06.007
  • 接收时间:2025-08-05
  • 首发时间:2026-08-05
  • 出版时间:2026-03-30
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  • 收稿日期:2025-08-05
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武汉市医学科学研究项目(WX23Z53)
作者信息
    1.武汉市第四医院古田院区急诊外科,武汉 430030
    2.武汉市第四医院古田院区急诊内科,武汉 430030

通讯作者:

李鸣(1970—),男,本科,主任医师,研究方向:肝胆、胃肠等及疾病的诊治及腹腔镜治疗。E-mail:
参考文献
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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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