Bone marrow-derived mesenchymal stem cells alleviate inflammation and tissue injury in severe acute pancreatitis by suppressing NLRP3/GSDMD-mediated pyroptosis

LI Yaxi; JIN Jing; ZHANG Yajing; MENG Huannan; ZHAO Hang

doi:10.16016/j.2097-0927.202511054

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Bone marrow-derived mesenchymal stem cells alleviate inflammation and tissue injury in severe acute pancreatitis by suppressing NLRP3/GSDMD-mediated pyroptosis

Basic Medicine | 更新时间：2026-02-23

- Bone marrow-derived mesenchymal stem cells alleviate inflammation and tissue injury in severe acute pancreatitis by suppressing NLRP3/GSDMD-mediated pyroptosis
- Journal of Army Medical University Vol. 48, Issue 3, Pages: 261-271(2026)
- 作者机构：
  
  1.上海中医药大学附属市中医医院消化内科，上海
  2.海军军医大学第一附属医院消化内科，上海
- 作者简介：
  
  ZHAO Hang， E-mail： drzhaohang@163.com
- 基金信息：
  
  the Open Project of State Key Laboratory of Immunology and Inflammation(2024Q0011)
- DOI：10.16016/j.2097-0927.202511054
  CLC： R329.21;R363.22;R576.05
- Received：10 November 2025，
  
  Revised：2026-01-12，
  
  Accepted：13 January 2026，
  
  Published：15 February 2026
- 稿件说明：
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李亚希，金晶，张雅婧，等.骨髓间充质干细胞通过抑制NLRP3/GSDMD介导的细胞焦亡减轻重症急性胰腺炎炎症与组织损伤［J］.陆军军医大学学报，2026，48（03）：261-271.DOI：10.16016/j.2097-0927.202511054.

LI Y X， JIN J，ZHANG Y J，et al.Bone marrow-derived mesenchymal stem cells alleviate inflammation and tissue injury in severe acute pancreatitis by suppressing NLRP3/GSDMD-mediated pyroptosis［J］.J Army Med Univ，2026，48（03）：261-271.DOI：10.16016/j.2097-0927.202511054.
李亚希，金晶，张雅婧，等.骨髓间充质干细胞通过抑制NLRP3/GSDMD介导的细胞焦亡减轻重症急性胰腺炎炎症与组织损伤［J］.陆军军医大学学报，2026，48（03）：261-271.DOI：10.16016/j.2097-0927.202511054. DOI：

LI Y X， JIN J，ZHANG Y J，et al.Bone marrow-derived mesenchymal stem cells alleviate inflammation and tissue injury in severe acute pancreatitis by suppressing NLRP3/GSDMD-mediated pyroptosis［J］.J Army Med Univ，2026，48（03）：261-271.DOI：10.16016/j.2097-0927.202511054. DOI：

摘要

目的

探索骨髓间充质干细胞（mesenchymal stem cells，MSCs）在抑制焦亡通路治疗重症急性胰腺炎（severe acute pancreatitis，SAP）大鼠模型的作用机制。

方法

冲洗骨髓腔法分离提取4周龄SPF级雄性SD大鼠［体质量（100±20）g］骨髓MSCs，并进行培养鉴定。将24只6~8周龄SPF级雄性大鼠［体质量（200±20）g］采用随机数字表法随机分为4组（

=6）：假手术组、SAP模型组、乌司他丁组、MSCs治疗组。SAP模型组、乌司他丁组、MSCs治疗组采用逆行胰胆管注射5%牛黄胆酸钠制备大鼠SAP模型，SAP模型组于造模后12 h注射100 μL PBS；乌司他丁组于模型制备完成后立即腹腔注射乌司他丁溶液（5×10

U/kg）；MSCs治疗组于造模后12 h注射MSCs细胞悬液（5×10

个/kg）；假手术组切开腹腔并数次翻动胰腺后，关闭腹腔。HE染色比较各组间胰腺组织形态，并进行病理学评分；试剂盒检测血清淀粉酶、脂肪酶水平；ELISA法检测血清炎症因子IL-10、IL-1β、TNF-α水平；透射电镜（transmission electron microscopy，TEM）观察胰腺腺泡细胞超微形态相关变化；免疫荧光观察胰腺组织NLRP3、GSDMD、caspase-1蛋白表达情况；qRT-PCR法检测胰腺组织NLRP3、GSDMD、caspase-1的mRNA含量；Western blot法检测胰腺组织NLRP3、GSDMD、GSDMD-N、caspase-1、cleaved caspase-1、IL-1β的蛋白相对表达量。

结果

骨髓MSCs具有成纤维细胞样纺锤体形状，并贴附在培养板的壁上；CD29（99.22%）呈高表达，CD45（0.42%）呈低

表达；具有成脂、成骨分化能力，细胞状态良好。与假手术组相比，SAP模型组在胰腺组织病理学评分、血清淀粉酶、脂肪酶、炎症因子等方面均有显著差异（

0.000 1）。与SAP模型组相比，乌司他丁组、MSCs治疗组胰腺组织病理学评分、血清淀粉酶、脂肪酶、IL-1β、TNF-α水平均降低（

0.001），IL-10水平升高（

0.05）；且MSCs治疗组上述指标较乌司他丁组明显好转（

0.05）。TEM显示MSCs治疗组腺泡细胞线粒体嵴恢复，线粒体形态改变，内质网肿胀减少；且与乌司他丁组相比，MSCs治疗组线粒体与内质网损伤程度减轻。免疫荧光结果显示，与SAP模型组相比，MSCs治疗组和乌司他丁组的胰腺组织NLRP3、GSDMD、caspase-1表达量下降（

0.000 1）。Western blot结果示，与SAP模型组相比，MSCs治疗组和乌司他丁组胰腺组织NLRP3、GSDMD、GSDMD-N、caspase-1、cleaved caspase-1、IL-1β蛋白相对表达量均有不同程度下降（

0.000 1）。qPR-PCR结果示，与SAP模型组相比，MSCs治疗组和乌司他丁组胰腺组织NLRP3、GSDMD、caspase-1 mRNA表达水平明显下降（

0.01）；且与乌司他丁组相比，MSCs治疗组上述指标均下降更为显著（

0.001）。

结论

MSCs减轻炎症反应，减少胰腺组织损伤，可通过抑制NLRP3/GSDMD/IL-1β通路改善腺泡细胞焦亡并缓解SAP。

Abstract

Objective

To investigate the mechanism of bone marrow-derived mesenchymal stem cells (MSCs) in alleviating severe acute pancreatitis (SAP) by inhibiting the pyroptosis pathway in a rat model.

Methods

Bone marrow-derived MSCs were isolated from 4-week-old male SPF SD rats [body weight

(100±20) g

]

using the bone marrow flush technique

followed by culture and characterization. Twenty-four male SPF SD rats [body weight (200±20) g

]

were randomly assigned to four groups (

=6 per group) using a random number table: a sham-operated group

an SAP model group

an ulinastatin-treated group

and an MSCs-treated group. SAP was induced in the latter three groups via retrograde injection of 5%sodium taurocholate into the biliopancreatic duct. The SAP model group received an injection of 100 μL PBS at 12 h post-modeling. The ulinastatin-treated group received an intraperitoneal injection of ulinastatin solution (5×10⁴ U/kg) immediately after model induction. The MSCs-treated group received an injection of MSC suspension (5×10⁶ cells/kg) at 12 h post-mode

ling. Sham-operated rats underwent laparotomy with gentle pancreatic manipulation before abdominal closure. Pancreatic histopathology was assessed by HE staining and pathological scoring. Serum amylase and lipase activities were measured using commercial assay kits. Serum levels of IL-10

IL-1β

and TNF-α were determined by ELISA. The ultrastructural changes of pancreatic acinar cells were observed under transmission electron microscopy (TEM). Protein expression of NLRP3

GSDMD

and caspase-1 in pancreatic tissue was evaluated by immunofluorescence. The mRNA levels of NLRP3

GSDMD

and caspase-1 were quantified by qRT-PCR. The relative protein expression levels of NLRP3

GSDMD

GSDMD-N

caspase-1

cleaved caspase-1

and IL-1β were analyzed by Western blotting.

Results

The isolated cells exhibited a typical adherent

spindle-shaped

fibroblast-like morphology. Flow cytometry confirmed they were highly positive for the mesenchymal marker CD29 (99.22%) and nearly negative for the hematopoietic marker CD45 (0.42%). These cells also demonstrated osteogenic and adipogenic differentiation potential and were maintained in good condition. Compared with the sham-operated group

the SAP model group showed significantly increased pancreatic histopathological scores

elevated serum amylase and lipase activities

higher levels of pro-inflammatory cytokines (IL-1β and TNF-α)

and a lower level of IL-10 (all

0.000 1). Both ulinastatin and MSCs treatment significantly attenuated these changes

reducing histopathological scores

serum enzyme activities

IL-1β

and TNF-α (

0.001)

while increasing IL-10 (

0.05). Importantly

the MSCs-treated group showed significantly greater improvement in all these metrics compared to the ulinastatin-treated group (

0.05). TEM analysis revealed that MSCs treatment promoted the recovery of mitochondrial cristae structure

improved overall organelle morphology

and reduced endopl

asmic reticulum swelling in acinar cells

with these restorative effects being more pronounced than in the ulinastatin group. The immunofluorescence results showed that compared with the SAP model group

the expression levels of NLRP3

GSDMD

and caspase-1 in pancreatic tissue were decreased in both the MSCs-treated group and the ulinastatin-treated group (

0.000 1). Western blot results indicated that

compared with the SAP model group

the relative protein expression levels of NLRP3

GSDMD

GSDMD-N

caspase-1

cleaved caspase-1

and IL-1β in pancreatic tissue were reduced to varying degrees in both the MSCs-treated group and the ulinastatin-treated group (

0.000 1). qPR-PCR results demonstrated that

compared with the SAP model group

the mRNA expression levels of NLRP3

GSDMD

and caspase-1 in pancreatic tissue were significantly decreased in both the MSCs-treated group and the ulinastatin-treated group (

0.01). Moreover

compared with the ulinastatin-treated group

the MSCs-treated group exhibited a more significant decrease in all the aforementioned indicators (

0.001).

Conclusion

Mesenchymal stem cells alleviate inflammation and tissue damage in severe acute pancreatitis. Their protective mechanism involves inhibiting the NLRP3/GSDMD/IL-1β signaling axis

thereby suppressing acinar cell pyroptosis.

关键词

Keywords

references

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