乳酸通过“心-胃轴”促进巨噬细胞 M1 型极化诱导胃黏膜损伤

doi:10.12122/j.issn.1673-4254.2026.06.07

摘要/Abstract

摘要：

目的探讨乳酸介导心肌缺血再灌注（MIR）诱导胃黏膜损伤的现象及机制。方法动物实验：30只C57BL/6J小鼠随机分为Sham组、MIR组（心肌缺血45 min，再灌注2 h）以及MIR+乳酸脱氢酶A抑制组（MIR+LDHAI组），每组10只。细胞实验：采用浓度梯度乳酸（0、5、15、30 mmol/L）刺激人单核细胞THP-1来源巨噬细胞极化；将人胃黏膜细胞 GES-1与乳酸或脂多糖（LPS）诱导的 M1型极化巨噬细胞间接共培养，并设置Control组（M0细胞上清）、LPS-M1组（含100 ng/mL LPS的M1细胞上清）以及Lactate-M1 组（含15 mmol/L乳酸的M1细胞上清）。采用胃黏膜损伤评定量化表进行胃黏膜大体观损伤量化评分；采用 HE染色观察心肌组织和胃黏膜组织病理改变；采用ELISA检测血清乳酸和cTnT水平；采用Western blotting和（或）免疫荧光法检测组织和（或）细胞中CD68、iNOS、Arg-1、COX-1和COX-2表达；采用代谢组学方法分析Sham和MIR两组血清代谢产物的差异。结果 MIR同时造成心脏和胃的损伤，表现为心肌组织水肿以及心肌细胞坏死，胃黏膜组织结构紊乱伴随免疫细胞浸润；Western blotting和免疫荧光染色显示MIR后胃黏膜组织iNOS蛋白水平增加，Arg-1蛋白表达降低，进一步采用荧光双重标记 CD68和iNOS，发现MIR后重叠的荧光区域增加，提示发生巨噬细胞M1型极化（P<0.05）。光镜下发现与M1型巨噬细胞间接共培养可诱导GES-1细胞损伤；Western blotting和免疫荧光染色提示M1细胞间接共培养诱导GES-1细胞COX-1蛋白表达减少，COX-2蛋白表达增加（P<0.05）。检测小鼠血清代谢组学提示MIR后循环乳酸的水平增加（P<0.05）。Western blotting和免疫荧光染色显示15 mmol/L乳酸诱导THP-1来源巨噬细胞M1型极化，表现为iNOS表达增加，Arg-1表达减少（P<0.05）；与乳酸和（或）LPS诱导的M1型巨噬细胞间接共培养诱导GES-1细胞损伤；抑制乳酸生成减轻MIR诱导的胃黏膜损伤，表现为胃黏膜组织结构改善和免疫细胞浸润减少。结论乳酸诱导巨噬细胞M1型极化参与MIR诱导的胃黏膜损伤，抑制乳酸生成可缓解乳酸通过“心-胃轴”造成的胃黏膜损伤。

关键词: 心肌缺血再灌注, 乳酸, 胃黏膜损伤, 巨噬细胞极化, 炎症损伤

Abstract:

Objective To investigate the role of lactate in myocardial ischemia-reperfusion (MIR)-induced gastric mucosal injury. Methods Thirty C57BL/6J mice were randomized into sham-operated group, MIR (45 min ischemia and 2 h reperfusion) group, and MIR+LDHAI group, and gastric mucosal injury was assessed using macroscopic scoring and HE staining; serum lactate and cTnT levels were measured using ELISA. Metabolomics analysis was conducted to identify the differential serum metabolites between the sham-operated and MIR mice. In the cell experiment, human gastric mucosal cells (GES-1) were indirectly co-cultured with THP-1 cell-derived macrophages polarized to the M1 phenotype by lactate and/or lipopolysaccharide (LPS) stimulation. In both the gastric mucosa tissues of the mice and the co-cultured GES-1 cells, the protein expression levels of CD68, iNOS, Arg-1, COX-1, and COX-2 were detected using Western blotting and immunofluo-rescence staining. Results Compared to sham operation, MIR induced concurrent injury in both the heart and stomach of the mice and caused elevation of iNOS protein levels and reduction of Arg-1 protein expression in the gastric mucosa, where increased overlapping fluorescent areas were detected after MIR using fluorescent double-labeling for CD68 and iNOS. Serum metabolomics analysis revealed a significant increase in circulating lactate levels after MIR. In GES-1 cells, indirect co-culture with M1 macrophages induced obvious cell injury, decreased COX-1 expression, and increased cellular COX-2 protein expression. Western blotting and immunofluorescence staining confirmed that 15 mmol/L lactate significantly increased iNOS and decreased Arg-1 expression in THP-1-derived macrophages. Indirect co-culture of GES-1 cells with M1 macrophages induced by lactate or LPS resulted in obvious GES-1 cell injury. In the mouse models of MIR, inhibition of lactic acid production effectively alleviated MIR-induced gastric mucosal injury. Conclusion Lactate-induced M1 macrophage polarization mediates MIR-related gastric mucosal injury in mice via the heart-gastric axis, an effect reversible by LDHA inhibition.

Key words: myocardial ischemia-reperfusion, lactate, gastric mucosal injury, macrophage polarization, inflammation

杨云恒, 贾雅清, 尹高生, 李双秀, 杨萍. 乳酸通过“心-胃轴”促进巨噬细胞 M1 型极化诱导胃黏膜损伤[J]. 南方医科大学学报, 2026, 46(6): 1267-1277.

Yunheng YANG, Yaqing JIA, Gaosheng YIN, Shuangxiu LI, Ping YANG. Lactate induces gastric mucosal injury by promoting M1 polarization of macrophages via the cardio-gastric axis[J]. Journal of Southern Medical University, 2026, 46(6): 1267-1277.

图/表 7

图1 小鼠 MIR 模型建立及鉴定

Fig1 Establishment and verification of the mouse model of myocardial ischemia-reperfusion (MIR). A: Schematic of the experimental protocol. C57 WT mice had an adaptation prior to MIR surgery on day 3. B: Electrocardiogram of the mice in each group. C: Pathological examination of gastric mucosa tissues in each group (HE staining) D: Level of serum cTnT in each group (n=10), respectively. *P<0.05.

图2 MIR 后小鼠胃黏膜组织病理改变

Fig.2 Histopathological changes in the gastric mucosa of the mice after MIR. A: Gastric mucosa of the mice after MIR. B: Gastric injury score of the mice in each group (****P<0.0001; n=10). C: HE staining of gastric mucosa tissues of the mice in each group.

图3 MIR小鼠胃黏膜组织iNOS表达增加, Arg-1表达降低

Fig.3 MIR induces upregulation of iNOS and downregulation of Arg-1 in the gastric mucosa of mice. A-C: Western Blotting for analyzing iNOS and Arg-1 expressions (**P<0.01,***P<0.001, n=10). D-G: Immunofluorescence staining images for analyzing iNOS and Arg-1 expressions (Original magnification: ×400; *P<0.05,**P<0.01; n=10). H-I: Immunofluorescent co-localization of CD68 and iNOS (×400; ****P<0.0001; n=10).

图4 LPS-M1 巨噬细胞诱导 GES-1 细胞细胞炎性损伤

Fig.4 LPS-M1 macrophages induce inflammatory injury in GES-1 cells. A: Microscopic observation of GES-1 cells before and after co-culture with M1 type macrophage (×200). B-D: Western blotting for analyzing COX-1 and COX-2 expressions. **P<0.01 (n=10).

图5 两组血清代谢组学比较

Fig.5 Comparative analysis of serum metabolomics between sham-operated and MIR mice. A: Scatter plot showing the differences between the two groups (OPLS-Damodel, n=5). B: Volcano plot showing the overall distribution of metabolite differences between the two groups. C: Distribution of major metabolites. D: Fold changes of some of upregulated and down-regulated metabolites after logarithmic transformation.

图6 体外乳酸诱导 THP-1 来源巨噬细胞 M1 型极化

Fig.6 Lactate-induced M1 polarization of THP-1 cells-derived macrophages in vitro. A-C: Western Blotting for analyzing iNOS and Arg-1 expressions (*P<0.05,**P<0.01,***P<0.001; n=10). D, E: Immunofluorescence staining for analyzing iNOS and Arg-1 expressions (×400). **P<0.01, ***P<0.001.

图8 抑制乳酸生成减轻MIR诱导的胃损伤

Fig.8 Reducing lactate alleviates gastric injury induced by MIR in mice. A: Schematic of the experimental protocol. C57 WT mice were given the LDHA inhibitor GSK2837808A (5 mg/kg) prior to MIR surgery on day 3. B: Level of serum lactate in MIR and MIR+LDHAI groups (**P<0.01; n=10). C: Gastric mucosa of the mice in MIR and MIR+LDHAI groups after MIR. D: Gastric injury score of the mice in each group (**P<0.01; n=10). E: HE staining of the gastric mucosa of the mice in MIR and MIR+LDHAI groups.

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