消退素D1在甲状旁腺激素促进牵张成骨中的水平变化及其对巨噬细胞极化效应研究

doi:10.13701/j.cnki.kqyxyj.2024.06.007

摘要/Abstract

摘要： 目的:研究甲状旁腺激素(parathyroid hormone, PTH)促进牵张成骨过程中消退素D1(resolvin D1, RvD1)表达水平变化及其对巨噬细胞极化的影响。方法:48只兔建立下颌骨牵张成骨模型,随机分为对照组和实验组。对照组术后注射生理盐水,实验组间歇性注射PTH 20 μg/kg。分别对各组兔牵张后第3、5、7天进行取样,通过苏木精-伊红(hematoxylin-eosin staining,HE)染色观察牵张区新骨组织;液相色谱串联质谱(liquid chromatography tandem mass spectrometry,LC-MS/MS)检测新骨组织中RvD1含量;实时荧光定量PCR(quantitative real-time PCR,qPCR)检测新骨组织中诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)及精氨酸酶1(Arginase 1,Arg1)的表达。体外构建由脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7炎症模型,并添加100 ng/mL RvD1进行处理,qPCR和流式细胞术检测巨噬细胞极化标记物的表达。结果:PTH促进下颌骨牵张成骨兔牵张区新骨生成并明显提高新骨中RvD1含量(P＜0.01)。同时,实验组牵张区新骨中巨噬细胞M2型极化标志物Arg1的表达升高,而iNOS的表达降低(P＜0.01)。体外实验结果显示RvD1上调了LPS处理的RAW264.7细胞中M2型极化标志物Arg1、CD206和抑炎因子白细胞介素-10(interleukin-10,IL-10)的表达(P＜0.05),并下调M1型标志物iNOS、CD86和促炎因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达(P＜0.01)。结论:PTH能提高兔下颌骨牵张新骨组织中RvD1含量,RvD1可进一步诱导巨噬细胞向M2型极化而发挥促进炎症消退和骨再生作用。

关键词: 消退素D1, 甲状旁腺激素, 牵张成骨, 巨噬细胞

Abstract: Objective: To study the changes of Resolvin D1 (RvD1) and its effect on macrophage polarization during mandibular distraction osteogenesis promoted by parathyroid hormone (PTH). Methods: The mandibular distraction osteogenesis model was established in 48 rabbits and the rabbits were randomly divided into two groups. The experimental group was injected with PTH 20 μg/kg intermittently after operation, while the control group was injected with normal saline. Samples were taken on the 3rd, 5th, and 7th day of distraction, respectively, and the new bone tissue in the distraction area was observed by hematoxylin-eosin (HE) staining. LC-MS/MS was used to detect the content of RvD1 in the new bone tissue in the distraction zone. qPCR was used to detect the mRNA level of iNOS and Arg1 in new bone tissue. RAW264.7 cells were cultured and treated with LPS, RvD1, and LPS+RvD1, respectively. The expression level of macrophage polarization markers was detected by qPCR and flow cytometry. Results: HE staining and gross specimen observation showed that the new bone formation in the distraction zone of the experimental group was better than that of the control group. The content of RvD1 in the experimental group was significantly higher than that in the control group (P<0.01). qPCR results showed that PTH significantly increased the expression level of M2-type polarization marker Arg1 gene in macrophages, and decreased the expression level of M1-type polarization marker iNOS (P<0.01). In vitro, the expression level of Arg1 mRNA of M2-type polarization marker was up-regulated after RvD1 treatment, while the expression of iNOS mRNA of M1-type polarization marker was down-regulated(P＜0.01). The results of flow cytometry showed that the LPS-induced increase in CD86 expression was significantly reduced and CD206 expression was significantly elevated after treatment of cells with RvD1. Conclusion: Intermittent injection of PTH can increase the content of RvD1 in rabbit mandibular distraction osteogenesis. RvD1 can polarize macrophages to M2, thus exerting anti-inflammatory effects.

Key words: Resolvin D1, parathyroid hormone, distraction osteogenesis, macrophage

刘毅恒, 田力月, 李永頔, 杨芷姗, 唐正龙. 消退素D1在甲状旁腺激素促进牵张成骨中的水平变化及其对巨噬细胞极化效应研究[J]. 口腔医学研究, 2024, 40(6): 506-511.

LIU Yiheng, TIAN Liyue, LI Yongdi, YANG Zhishan, TANG Zhenglong. Changes of Resolvin D1 and Its Polarization Effect on Macrophages in Mandibular Distraction Osteogenesis Promoted by Parathyroid Hormone Administration[J]. Journal of Oral Science Research, 2024, 40(6): 506-511.

参考文献

[1] 覃祥城,曾景奇,王凡,等.中药促进牵张成骨的研究进展[J].广州中医药大学学报, 2020, 37(2): 382-385.
[2] Inada N, Ohata T, Maruno H, et al. Optimal timing for intermittent administration of parathyroid hormone (1-34) for distraction osteogenesis in rabbits [J]. J Orthop Surg Res, 2022, 17(1): 130.
[3] Zhang W, Wang N, Yang M, et al. Periosteum and development of the tissue-engineered periosteum for guided bone regeneration [J]. J Orthop Translat, 2022, 33: 41-54.
[4] Zhao H, Jia Y, Wang F, et al. Cobalt-doped mesoporous silica coated magnetic nanoparticles promoting accelerated bone healing in distraction osteogenesis [J]. Int J Nanomedicine, 2023, 18: 2359-2370.
[5] Chen T, Wang Y, Hao Z, et al. Parathyroid hormone and its related peptides in bone metabolism [J]. Biochem Pharmacol, 2021, 192: 114669.
[6] 李永頔,朱鹏娜,王冬香,等.骨保护素/核因子κB受体活化因子配体在甲状旁腺激素调控下颌骨牵张成骨中的表达效应[J].华西口腔医学杂志,2016,34(3): 234-238.
[7] Ai-Aql ZS, Alagl AS, Graves DT, et al. Molecular mechanisms controlling bone formation during fracture healing and distraction osteogenesis [J]. J Dent Res, 2008, 87(2): 107-118.
[8] Zweifler LE, Sinder BP, Stephan C, et al. Parathyroid hormone and trabectedin have differing effects on macrophages and stress fracture repair [J]. Bone, 2024, 179: 116983.
[9] 袁红梅,万敬员,张力.促炎症消退新介质:消退素与保护素[J]. 生命科学,2012,24(1): 54-57.
[10] Xiang SY, Ye Y, Yang Q, et al. RvD1 accelerates the resolution of inflammation by promoting apoptosis of the recruited macrophages via the ALX/FasL-FasR/caspase-3 signaling pathway [J]. Cell Death Discov, 2021, 7(1): 339.
[11] Gao J, Su Y, Wang Z. Lung inflammation resolution by RvD1 and RvD2 in a receptor-dependent manner [J]. Pharmaceutics, 2023, 15(5):1527.
[12] 刘音辰.特异性促炎症消退介质在牙周炎中的研究进展[J].华西口腔医学杂志,2021,39(1): 94-98.
[13] Sordi R, Chiazza F, Collotta D, et al. Resolvin D1 attenuates the organ injury associated with experimental hemorrhagic shock [J]. Ann Surg, 2021, 273(5): 1012-1021.
[14] McCauley LK, Dalli J, Koh AJ, et al. Cutting edge: Parathyroid hormone facilitates macrophage efferocytosis in bone marrow via proresolving mediators resolvin D1 and resolvin D2 [J]. J Immunol, 2014, 193(1): 26-29.
[15] Tang ZL, Zhang WJ, Wang DX, et al. An experimental study addressing the promotion of mandibular defect repair through the intermittent subcutaneous injection of parathyroid hormone [J]. J Oral Maxillofac Surg, 2014, 72(2): 419-430.
[16] Ohata T, Maruno H, Ichimura S. Changes over time in callus formation caused by intermittently administering PTH in rabbit distraction osteogenesis models [J]. J Orthop Surg Res, 2015, 10: 88.
[17] Wang SP, Chen YJ, Hsu CE, et al. Intermittent parathyroid hormone treatment affects the bone structural parameters and mechanical strength of the femoral neck after ovariectomy-induced osteoporosis in rats [J]. Biomed Eng Online, 2022, 21(1): 6.
[18] Yamashita J, McCauley LK. Effects of intermittent administration of parathyroid hormone and parathyroid hormone-related protein on fracture healing: A narrative review of animal and human studies [J]. JBMR Plus, 2019, 3(12): e10250.
[19] Wagner F, Vach W, Augat P, et al. Daily subcutaneous Teriparatide injection increased bone mineral density of newly formed bone after tibia distraction osteogenesis, a randomized study [J]. Injury, 2019, 50(8): 1478-1482.
[20] Shen H, Jiang W, Yu Y, et al. microRNA-146a mediates distraction osteogenesis via bone mesenchymal stem cell inflammatory response [J]. Acta Histochem, 2022, 124(6): 151913.
[21] Zhang J, Chen J, Jiang Q, et al. Resolvin D1 attenuates inflammation and pelvic pain associated with EAP by inhibiting oxidative stress and NLRP3 inflammasome activation via the Nrf2/HO-1 pathway [J]. J Inflamm Res, 2023, 16: 3365-3379.
[22] Xu B, Li M, Cheng T, et al. Resolvin D1 protects against sepsis-associated encephalopathy in mice by inhibiting neuro-inflammation induced by microglia [J]. Am J Transl Res, 2022, 14(9): 6737-6750.
[23] Pinto N, Klein Y, David E, et al. Resolvin D1 improves allograft osteointegration and directly enhances osteoblasts differentiation [J]. Front Immunol, 2023, 14: 1086930.
[24] Benabdoun HA, Kulbay M, Rondon EP, et al. In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis [J]. Arthritis Res Ther, 2019, 21(1): 72.
[25] Runtsch MC, Angiari S, Hooftman A, et al. Itaconate and itaconate derivatives target JAK1 to suppress alternative activation of macrophages [J]. Cell Metab, 2022, 34(3): 487-501.e8.
[26] Muñoz J, Akhavan NS, Mullins AP, et al. Macrophage polarization and osteoporosis: A review [J]. Nutrients, 2020, 12(10):2999.
[27] Chazaud B. Inflammation and skeletal muscle regeneration: Leave it to the macrophages! [J]. Trends Immunol, 2020, 41(6): 481-492.
[28] 聂张玲,包崇云.巨噬细胞影响骨形成机制的研究进展[J].口腔医学研究,2021,37(11): 966-969.