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PKM2调控的巨噬细胞糖酵解在糖尿病肾病炎症中的作用
Role of pyruvate kinase M20-regulated glycolysis in inflammatory activation of macrophages in diabetic nephropathy
- 第三军医大学学报 2020年42卷第24期 页码：2394-2402
- 作者机构：
1.230031 合肥，安徽医科大学第四附属医院肾内科
2.230031 合肥，安徽医科大学第一附属医院肾内科
- 作者简介：
[ 徐兴欣，E-mail:xuxingxin.2008@163.com XU Xingxin, E-mail:xuxingxin.2008@163.com
- 基金信息：国家自然科学基金青年科学基金(81600568);安徽省自然科学基金青年项目(1708085QH178)
DOI：10.16016/j.1000-5404.202007015
中图分类号： R392.32;R587.2;R692收稿：2020-07-02，
修回：2020-09-10，
纸质出版：2020-12-30
- 稿件说明：
移动端阅览
谢姗, 徐兴欣. PKM2调控的巨噬细胞糖酵解在糖尿病肾病炎症中的作用[J]. 第三军医大学学报, 2020,42(24):2394-2402.
Shan XIE, Xingxin XU. Role of pyruvate kinase M20-regulated glycolysis in inflammatory activation of macrophages in diabetic nephropathy[J]. Journal of Third Military Medical University, 2020, 42(24): 2394-2402.
谢姗, 徐兴欣. PKM2调控的巨噬细胞糖酵解在糖尿病肾病炎症中的作用[J]. 第三军医大学学报, 2020,42(24):2394-2402. DOI： 10.16016/j.1000-5404.202007015.
Shan XIE, Xingxin XU. Role of pyruvate kinase M20-regulated glycolysis in inflammatory activation of macrophages in diabetic nephropathy[J]. Journal of Third Military Medical University, 2020, 42(24): 2394-2402. DOI： 10.16016/j.1000-5404.202007015.
摘要
目的
2
探究在糖尿病肾病(diabetic nephropathy
DN)中M2型丙酮酸激酶(pyruvate kinase M2
PKM2)调控的糖酵解对Nod样受体蛋白3(nod-like receptor protein 3
NLRP3)和黑色素瘤缺乏因子2(absentin melanoma 2
AIM2)介导的巨噬细胞炎症激活的影响。
方法
2
① 收取30例DN患者的血清和肾穿刺活检标本作为DN组，20例健康人血清标本和20例癌旁正常肾组织标本作为正常(Normal)组，通过免疫组化检测糖酵解标志物PKM2、巨噬细胞标志物CD68、炎症因子TNF-α的表达变化，ELISA检测血清TNF-α、IL-1β、MCP-1水平，激光共聚焦显微镜观察肾组织CD68与PKM2共表达。②选取小鼠骨髓来源巨噬细胞分组培养，通过简单随机化的方法分为甘露醇对照组、siRNA对照组、低糖对照组、高糖刺激组、高糖刺激+ PKM2 siRNA组、高糖刺激+糖酵解抑制剂组；Western blot检测各组细胞PKM2、p-EIF2AK2、EIF2AK2、NLRP3、AIM2表达，ELISA及RT-PCR检测各组细胞MCP-1、IL-1β、TNF-α分泌及mRNA表达，乳酸测定试剂盒及葡萄糖吸收试剂盒检测各组细胞乳酸含量和葡萄糖吸收。
结果
2
与Normal组相比，DN患者的肾组织中PKM2表达增加(
P
<
0.01)，巨噬细胞浸润增多及炎症因子表达增多(
P
<
0.01)。与Control组相比，高糖可以上调巨噬细胞PKM2、p-EIF2AK2、NLRP3、AIM2蛋白表达(
P
<
0.01)及MCP-1、IL-1β、TNF-α mRNA的表达(
P
<
0.01)，增加细胞上清液中MCP-1、IL-1β、TNF-α、乳酸及葡萄糖水平(
P
<
0.01)，而PKM2 siRNA可以抑制高糖诱导的炎症。
结论
2
高糖可以刺激巨噬细胞的糖酵解水平增加，并促进炎症小体激活及炎症因子的释放，其机制与PKM2- EIF2AK2信号通路激活有关。
Abstract
Objective
2
To explore the role of pyruvate kinase M20 (PKM2)-regulated glycolysis on nod-like receptor protein 3 (NLRP3)- and absentin melanoma 2 (AIM2)-mediated inflammatory activation of macrophages in diabetic nephropathy (DN).
Methods
2
Renal biopsy samples from 30 DN patients and tumor-adjacent renal tissue samples from 20 renal cancer patients were collected for detection of the expressions of the glycolytic marker PKM2
the macrophage infiltration marker CD68 and tumor necrosis factor-α (TNF-α) using immunohistochemistry. The levels of TNF-α
interleukin-1β (IL-1β) and monocyte chemoattractant protein-1 (MCP-1) in serum samples from the DN patients and 20 healthy volunteers were detected by ELISA. In cultured mouse bone marrow-derived macrophages
the effects of mannitol
high-glucose exposure
high-glucose exposure plus PKM2 siRNA
and high-glucose exposure plus glycolysis inhibitor on the expression of PKM2
p-EIF2AK2
EIF2AK2
NLRP3 and AIM2 were detected using Western blotting; the secretion and mRNA expression of TNF-α
IL-1β and MCP-1 were detected by ELISA and RT-PCR
and the lactate content and glucose absorption of the cells were assayed using commercial detection kits.
Results
2
Compared with the tumor-adjacent renal tissues
the renal tissues from DN patients showed significantly increased expression of PKM2
CD68 and TNF-α (
P
<
0.01). In the cell cultures of macrophages
high glucose exposure obviously upregulated the cellular expression of PKM2
p-EIF2AK2
NLRP3 and AIM2 proteins (
P
<
0.01) and the mRNA expression of TNF-α
IL-1β and MCP-1 (
P
<
0.01) and increased the levels of TNF-α
IL-1β
MCP-1
lactic acid and glucose in the cell supernatant (
P
<
0.01). Transfection of the cells with PKM2 siRNA significantly inhibited inflammatory responses of the macrophages induced by high-glucose exposure.
Conclusion
2
High glucose stimulation can enhance the level of glycolysis in macrophages and promote the activation of inflammasomes and release of inflammatory cytokines through a mechanism involving the activation of PKM2-EIF2AK2 signaling pathway.
关键词
Keywords
references
FU J, AKAT K M, SUN Z, et al . Single-cell RNA profiling of glomerular cells shows dynamic changes in experimental diabetic kidney disease[J ] . J Am Soc Nephrol, 2019, 30(4): 533-545. DOI: 10.1681/asn.2018090896.
BHATT K, LANTING L L, JIA Y, et al . Anti-inflammatory role of MicroRNA-146a in the pathogenesis of diabetic nephropathy[J ] . J Am Soc Nephrol, 2016, 27(8): 2277-2288. DOI: 10.1681/asn.2015010111.
ANGIARI S, RUNTSCH M C, SUTTON C E, et al . Pharmacological activation of pyruvate kinase M2 inhibits CD4 + T cell pathogenicity and suppresses autoimmunity[J ] . Cell Metab, 2020, 31(2): 391-405.e8. DOI: 10.1016/j.cmet.2019.10.015..
XIE M, YU Y, KANG R, et al . PKM2-dependent glycolysis promotes NLRP3 and AIM2 inflammasome activation[J ] . Nat Commun, 2016, 7: 13280. DOI: 10.1038/ncomms13280.
鞠小丽, 王强. EIF2AK2和NLRP3相互作用影响细胞分泌IL-1β的研究[J].安徽医科大学学报, 2016, 51(11): 1565-1568.
JU X L, WANG Q. Investigation of EIF2AK2 interaction with NLRP3 and effect on the release of IL-1β[J]. Acta Univ Med Anhui, 2016, 51(11): 1565-1568.
CHOI S Y, LIM S W, SALIMI S, et al . Tonicity-responsive enhancer-binding protein mediates hyperglycemia-induced inflammation and vascular and renal injury[J ] . J Am Soc Nephrol, 2018, 29(2): 492-504. DOI: 10.1681/asn.2017070718.
LINKERMANN A, PARMENTIER S P, HUGO C. We AIM2 inflame[J]. J Am Soc Nephrol, 2018, 29(4): 1077-1079. DOI: 10.1681/asn.2018020116.
PALSSON-MCDERMOTT E M, CURTIS A M, GOEL G, et al . Pyruvate kinase M2 regulates Hif-1α activity and IL-1β induction and is a critical determinant of the Warburg effect in LPS-activated macrophages[J ] . Cell Metab, 2015, 21(1): 65-80. DOI: 10.1016/j.cmet.2014.12.005.
CALLE P, HOTTER G. Macrophage phenotype and fibrosis in diabetic nephropathy[J]. Int J Mol Sci, 2020, 21(8): 2806. DOI: 10.3390/ijms21082806.
PRÓCHNICKI T, LATZ E. Inflammasomes on the crossroads of innate immune recognition and metabolic control[J]. Cell Metab, 2017, 26(1): 71-93. DOI: 10.1016/j.cmet.2017.06.018.
CORCORAN S E, O'NEILL L A. HIF1α and metabolic reprogramming in inflammation[J]. J Clin Invest, 2016, 126(10): 3699-3707. DOI: 10.1172/jci84431.
周赟, 易竹君, 阙克婷, 等.铁离子通过ROS-乙酰化P53促进巨噬细胞M1型极化[J].第三军医大学学报, 2018, 40(1): 45-52.
ZHOU Y, YI Z J, QUE K T, et al . Iron ion polarizes macrophages to M1 subtype through ROS-acetyl-P53[J ] . J Third Mil Med Univ, 2018, 40(1): 45-52.
HUANG J, LIU K, ZHU S, et al . AMPK regulates immunometabolism in Sepsis[J ] . Brain Behav Immun, 2018, 72: 89-100. DOI: 10.1016/j.bbi.2017.11.003.
CHO S J, MOON J S, NIKAHIRA K, et al . GLUT1-dependent glycolysis regulates exacerbation of fibrosis via AIM2 inflammasome activation[J ] . Thorax, 2020, 75(3): 227-236. DOI: 10.1136/thoraxjnl-2019-213571.
HUGHES M M, O'NEILL L A J. Metabolic regulation of NLRP3[J]. Immunol Rev, 2018, 281(1): 88-98. DOI: 10.1111/imr.12608.
YANG L, XIE M, YANG M, et al . PKM2 regulates the Warburg effect and promotes HMGB1 release in sepsis[J ] . Nat Commun, 2014, 5: 4436. DOI: 10.1038/ncomms5436.
ERRA DÍAZ F, DANTAS E, GEFFNER J. Unravelling the interplay between extracellular acidosis and immune cells[J]. Mediators Inflamm, 2018, 2018: 1218297. DOI: 10.1155/2018/1218297.
LU B, NAKAMURA T, INOUYE K, et al . Novel role of PKR in inflammasome activation and HMGB1 release[J ] . Nature, 2012, 488(7413): 670-674. DOI: 10.1038/nature11290.
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