Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway
ZHONG Chang-rui1, FU Chun-hua2
1. Longyan First Affiliated Hospital of Fujian Medical University, Longyan Fujian 364000, China; 2. The Second Hospital of Longyan, Longyan Fujian 364000, China
Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway
ZHONG Chang-rui1, FU Chun-hua2
1. Longyan First Affiliated Hospital of Fujian Medical University, Longyan Fujian 364000, China; 2. The Second Hospital of Longyan, Longyan Fujian 364000, China
摘要Objective: To explore the effect of platelet-rich plasma(RPR) in stimulating the transformation of pro-inflammatory(M1) macrophages into antiinflammatory(M2) under the adenosine-monophosphate-dependent protein kinase(AMPK) signaling pathway. Methods:Rat peritoneal macrophages(RAW264.7) were cultured and randomly divided into 8 groups: blank control group, LPS group, RPR group A, RPR group B, LPS+RPR(12 h) group, LPS+RPR(24 h) group A, LPS+RPR(24 h) group B, LPS+RPR(24 h) group C. RPR was prepared based on blood donors. The expressions of AMPK signaling pathway-related proteins(AMPK, ULK1, mTOR) and macrophage markers(iNOS, Arg-1) in the blank control group, LPS group, LPS+RPR(12 h) group and LPS+RPR(24 h) group were observed and compared. The expressions of macrophage markers in LPS+RPR(24 h) B and C groups were compared, and the expressions of AMPK and TGF-β in RPR A and B groups were compared. Results: The gray values of AMPK and ULK1 in LPS cells decreased significantly, while those in LPS+RPR(12 h) and LPS+RPR(24 h) A cells increased significantly. The gray values of AMPK and ULK1 in LPS+RPR(24 h) A cells were higher than those in LPS+RPR(12 h) cells(P<0.05). The mTOR gray value of LPS cells was significantly higher than that of LPS+RPR(24 h) A cells, and the mTOR gray value of LPS+RPR(24 h) A cells was significantly lower than that of LPS+RPR(12 h) cells(P<0.05). The expression of iNOS in LPS cells was significantly decreased, the expression of iNOS in LPS+RPR(12 h) and LPS+RPR(24 h) cells was significantly increased, and the expression of iNOS in LPS+RPR(24 h) cells was higher than that in LPS+RPR(12 h) cells(P<0.05). The expression of Arg-1 in LPS cells was significantly decreased, the expression of Arg-1 in LPS+RPR(12 h) and LPS+RPR(24 h) A cells was significantly increased, and the expression of Arg-1 in LPS+RPR(24 h) A cells was higher than that in LPS+RPR(12 h) cells(P<0.05). The iNOS expression level of LPS+PRP(24 h) C cells was significantly higher than that of LPS+PRP(24 h) B cells, and the Arg-1 expression level was significantly lower than that of LPS+PRP(24 h) B cells(P<0.05). The gray values of AMPK and TGF-β in PRP B cells were significantly lower than those in PRP A cells(P<0.05). Conclusion: RPR can stimulate macrophage transformation from M1 to M2 by up-regulating AMPK signaling pathway.
Abstract：Objective: To explore the effect of platelet-rich plasma(RPR) in stimulating the transformation of pro-inflammatory(M1) macrophages into antiinflammatory(M2) under the adenosine-monophosphate-dependent protein kinase(AMPK) signaling pathway. Methods:Rat peritoneal macrophages(RAW264.7) were cultured and randomly divided into 8 groups: blank control group, LPS group, RPR group A, RPR group B, LPS+RPR(12 h) group, LPS+RPR(24 h) group A, LPS+RPR(24 h) group B, LPS+RPR(24 h) group C. RPR was prepared based on blood donors. The expressions of AMPK signaling pathway-related proteins(AMPK, ULK1, mTOR) and macrophage markers(iNOS, Arg-1) in the blank control group, LPS group, LPS+RPR(12 h) group and LPS+RPR(24 h) group were observed and compared. The expressions of macrophage markers in LPS+RPR(24 h) B and C groups were compared, and the expressions of AMPK and TGF-β in RPR A and B groups were compared. Results: The gray values of AMPK and ULK1 in LPS cells decreased significantly, while those in LPS+RPR(12 h) and LPS+RPR(24 h) A cells increased significantly. The gray values of AMPK and ULK1 in LPS+RPR(24 h) A cells were higher than those in LPS+RPR(12 h) cells(P<0.05). The mTOR gray value of LPS cells was significantly higher than that of LPS+RPR(24 h) A cells, and the mTOR gray value of LPS+RPR(24 h) A cells was significantly lower than that of LPS+RPR(12 h) cells(P<0.05). The expression of iNOS in LPS cells was significantly decreased, the expression of iNOS in LPS+RPR(12 h) and LPS+RPR(24 h) cells was significantly increased, and the expression of iNOS in LPS+RPR(24 h) cells was higher than that in LPS+RPR(12 h) cells(P<0.05). The expression of Arg-1 in LPS cells was significantly decreased, the expression of Arg-1 in LPS+RPR(12 h) and LPS+RPR(24 h) A cells was significantly increased, and the expression of Arg-1 in LPS+RPR(24 h) A cells was higher than that in LPS+RPR(12 h) cells(P<0.05). The iNOS expression level of LPS+PRP(24 h) C cells was significantly higher than that of LPS+PRP(24 h) B cells, and the Arg-1 expression level was significantly lower than that of LPS+PRP(24 h) B cells(P<0.05). The gray values of AMPK and TGF-β in PRP B cells were significantly lower than those in PRP A cells(P<0.05). Conclusion: RPR can stimulate macrophage transformation from M1 to M2 by up-regulating AMPK signaling pathway.
ZHONG Chang-rui, FU Chun-hua. Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway[J]. 中国生物医学工程学报(英文版）, 2024, 33(2): 85-92.
ZHONG Chang-rui, FU Chun-hua. Effect of Platelet-rich Plasma in Stimulating Macrophage Transformation into M2 Type under AMPK Signaling Pathway. Chinese Journal of Biomedical Engineering, 2024, 33(2): 85-92.
[1] Shi Zhongmin, Gu Wenqi, Yang Yunfeng, et al.Expert consensus on platelet-rich plasma treatment for osteochondral lesion of talus (version 2023)[J].Chinese Journal of Trauma,2023,39(5): 385-393. [2] Andia I, Abate M.Platelet-rich plasma: underlying biology and clinical correlates[J]. Regen Med, 2013, 8(5): 645-658. [3] Sacta MA, Tharmalingam B, Coppo M, et al.Gene-specific mechanisms direct glucocorticoid-receptor-driven repression of inflammatory response genes in macrophages[J]. Elife, 2018, 7: e34864. [4] Jia Rui, Hui Yi, Yan Shuguang, et al.Research progress on relationship between macrophage M1/M2 polarization and immune inflammatory diseases[J]. Chinese Journal of Immunology,2021, 37(22): 2791-2797. [5] Kohlstedt K, Trouvain C, Namgaladze D, et al.Adipocyte-derived lipids increase angiotensin-converting enzyme (ACE) expression and modulate macrophage phenotype[J]. Basic Res Cardiol, 2011, 106(2): 205-215. [6] Hu L, Shan Z, Wang F, et al.Vascular endothelial growth factor B exerts lipid-lowering effect by activating AMPK via VEGFR1[J]. Life Sci, 2021, 276: 119401. [7] Saucedo JM, Yaffe MA, Berschback JC, et al.Platelet-rich plasma[J]. J Hand Surg Am, 2012, 37(3): 587-589. [8] Wang Li, Zhang Mengting, Chen Yang, et al.Effect of human amniotic epithelial cells combined with platelet rich plasma on fibrotic factors and inflammatory factors in rats with intrauterine adhesions[J]. Progress in Modern Biomedicine, 2023, 23(8): 1430-1435. [9] Xiang HC, Lin LX, Hu XF, et al.AMPK activation attenuates inflammatory pain through inhibiting NF-κB activation and IL-1β expression[J]. J Neuroinflammation, 2019, 16(1): 34. [10] Namgaladze D, Snodgrass RG, Angioni C, et al.AMP-activated protein kinase suppresses arachidonate 15-lipoxygenase expression in interleukin 4-polarized human macrophages[J]. J Biol Chem, 2015, 290(40): 24484-24494. [11] Shi Linying, Li Yanhui, Xu Jingjing, et al.Platelet-rich plasma induces M2 macrophage polarization via regulating AMPK signaling pathway[J].Journal of Experimental Hematology, 2023, 31(5): 1486-1491.
