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目的 研究药桑提取物脱氧野尻霉素(1-Deoxynojirimycin, DNJ)在高糖环境下对MC3T3-E1细胞的增殖和成骨分化的作用。方法 取P4代MC3T3-E1细胞，在50 mmol/L的高糖环境下通过细胞毒性检测试剂盒(Cell dountingkit-8,CCK-8)检测不同浓度的DNJ(10 mmol/L、1 mmol/L、100μmol/L、10μmol/L、1μmol/L、100 nmol/L、10 nmol/L、1 nmol/L)干预对MC3T3-E1细胞增殖的影响，筛选出后续实验研究的DNJ浓度。取培养至对数生长期的MC3T3-E1细胞，按不同干预方式分为：空白组(完全培养基)、高糖组(50 mmol/L糖浓度的完全培养基)、实验组[50 mmol/L糖浓度的完全培养基+不同浓度DNJ(100、10、1μmol/L)溶液]。使用流式细胞技术检测细胞的凋亡和活性氧，采用ELISA试剂盒检测细胞上清液中AGEs和IL-1β、IL-6、TNF-α的含量，通过碱性磷酸酶染色及活性检测MC3T3-E1细胞的早期成骨能力的影响，通过茜素红染色和定量检测MC3T3-E1细胞的晚期成骨能力，采用RT-PCR检测IL-1β、IL-6、TNF-α、Bax、Bcl-2、IGF-1、ALP、OCN、OSX、Col-1和Runx2的mRNA表达情况。结果 高糖环境下，100、10、1μmol/L的DNJ可以促进MC3T3-E1细胞增殖。与高糖组相比，实验组凋亡率、活性氧含量、细胞上清液中AGEs、IL-1β、IL-6和TNF-α含量降低，差异有统计学意义(P<0.05)。RT-PCR结果表明，与高糖组相比，实验组Bax、IL-1β、IL-6和TNF-α的mRNA表达降低，Bcl-2、IGF-1、ALP、OCN、OSX、Col-1和Runx2的mRNA表达升高，差异有统计学意义(P<0.05)。结论 在高糖环境下，一定浓度范围内DNJ能够抑制MC3T3-E1细胞的凋亡和炎症因子的产生，促进成骨细胞的分化。
Abstract：Objective To investigate the effect of 1-Deoxynojirimycin(DNJ) on the proliferation and osteogenic differentiation of MC3T3-E1 cells in high glucose environment. Methods The P4 generation MC3T3-E1 cells were taken, and the effects of different concentrations of DNJ(10 mmol/L, 1 mmol/L,100 μmol/L, 10 μmol/L, 1 μmol/L, 100 nmol/L, 10 nmol/L and 1 nmol/L) on the proliferation of osteogenic precursor cells were detected by CellDounting Kit-8(CCK-8) in a high glucose environment of 50 mmol/L, and the concentration of DNJ in subsequent experimental studies was screened. MC3T3-E1 cells cultured to logarithmic growth phase were selected and divided into blank group(complete medium), high glucose group(complete medium with 50 mmol/L glucose concentration) and experimental group [complete medium with 50 mmol/L glucose concentration+different concentrations of DNJ(100 μmol/L, 10 μmol/L, 1 μmol/L) solution]. Flow cytometry was used to detect cell apoptosis and reactive oxygen species production. The contents of AGEs, IL-1β, IL-6 and TNF-α were detected by ELISA kit. The early osteogenic ability of MC3T3-E1 cells were detected by alkaline phosphatase staining and activity. The late osteogenic ability of MC3T3-E1 cells were detected by alizarin red staining and quantitative detection. The mRNA expressions of IL-1β, IL-6, TNF-α, Bax, Bcl-2, IGF-1, ALP, OCN, OSX, Col-1 and Runx2 were detected by RT-PCR. Results In the high glucose environment, 100 μmol/L, 10 μmol/L and 1 μmol/L DNJ can significantly promote the proliferation of MC3T3-E1 cells, so it is used for subsequent research. Compared with the high glucose group, the apoptosis rate, reactive oxygen species and the contents of AGEs, IL-1β, IL-6 and TNF-α in the cell supernatant of the experimental group weredecreased, and the difference was statistically significant(P<0.05). The results of RT-PCR showed that compared with the high glucose group, the mRNA expression of Bax, IL-1β, IL-6 and TNF-α in the experimental group was significantly decreased, and the mRNA expression of Bcl-2, IGF-1, ALP, OCN, OSX, Col-1 and Runx2 was significantly increased, the difference was statistically significant(P<0.05). Conclusion In the high glucose environment, DNJ can inhibit the apoptosis of mouse pre-osteoblast MC3T3-E1 and the production of inflammatory factors within a certain concentration range, and promote the differentiation of osteoblasts.
[1] LATUSEK K,S?OTWINSKA-PAWLACZYK A,WARAKOMSKA A,et al.Pilot study:the effectiveness of hyperbaric oxygen therapy in the treatment of periodontitis in patients with type 2 diabetes[J].Healthcare (Basel),2023,11(9):1344-1355.
[2] WU C Z,YUAN Y H,LIU H H,et al.Epidemiologic relationship between periodontitis and type 2 diabetes mellitus[J].BMC Oral Health,2020,20(1):204-219.
[3] PRESHAW P M,BISSETT S M.Periodontitis and diabetes[J].Br Dent J,2019,227(7):577-584.
[4] TAN L,LIU J,LIU Z.Association between periodontitis and the prevalence and prognosis of prediabetes:a population-based study[J].Transl Med,2023,21(1):484-494.
[5] AUSENDA F,BARBERA E,COTTI E,et al.Clinical,microbiological and immunological short,medium and long-term effects of different strains of probiotics as an adjunct to non-surgical periodontal therapy in patients with periodontitis systematic review with meta-analysis[J].Jpn Dent Sci Rev,2023,59(4):62-73.
[6] MOHSENR O M,HASSAN R.A comparative study of the therapeutic effect of bone marrow mesenchymal stem cells versus insulin on mandibular dento-alveolar complex collagen formation and beta-catenin expression in experimentally induced type I diabetes[J].Saudi Dent,2023,35(6):668-677.
[7] DU Y,LI D X,LU D Y,et al.Amelioration of lipid accumulations and metabolism disorders in differentiation and development of 3T3-L1 adipocytes through mulberry leaf water extract[J].Phytomedicine,2022,98(1):153959-153964.
[8] 王诗睿，赵冲.桑叶中1-脱氧野尻霉素降糖机制研究进展[J].核农学报，2023,37(11):2252-2257.
[9] 张文杰，吴泽钰，夏雨凝，等.桑叶水提物对高糖环境下前成骨细胞成骨分化的作用研究[J].现代口腔医学杂志，2024,38(1):1-8.
[10] PANG Y,YUAN X,GUO J,et al.The effect of liraglutide on the proliferation,migration,and osteogenic differentiation of human periodontal ligament cells[J].Periodontal Res,2019,54(2):106-114.
[11] REN W,CHAI M,JIANG M,et al.High glucose mediates apoptosis and osteogenesis of MSCs via downregulation of AKT-Sirt1-TWIST[J].Mol Biol Rep,2022,49(4):2723-2733.
[12] 李军，靳爽，张娟，等.利拉鲁肽在高糖环境下对体外培养的小鼠前成骨细胞MC3T3-E1增殖、分化的影响[J].中华内分泌代谢杂志，2016,32(3):232-235.
[13] JIN Q H,HU X J,ZHAO H Y.Curcumin activates autophagy and attenuates high glucose-induced apoptosis in HUVECs through the ROS/NF-κB signaling pathway[J].Experimental Therapeutic Medicine,2022,24(3):596-606.
[14] LI Z,LI S,WANG N,et al.Liraglutide,a glucagon-like peptide-1 receptor agonist,suppresses osteoclastogenesis through the inhibition of NF-κB and MAPK pathways via GLP-1R[J].Biomed Pharmacother,2020,130(38):110523-110533.
[15] LUC K,SCHRAMM-LUC A,GUZIK T J,et al.Oxidative stress and inflammatory markers in prediabetes and diabetes[J].Physiol Pharmacol,2019,70(6):10-26.
[16] SRUTHI C R,RAGHU K G.Advanced glycation end products and their adverse effects:The role of autophagy[J].Biochem Mol Toxicol,2021,35(4):22710-22727.
[17] CIPRIANI C,COLANGELO L,SANTORI R,et al.The interplay between bone and glucose metabolism[J].Front Endocrinol (Lausanne),2020,24(11):122-131.
[18] YAN Y,ZHANG H,LIU L,et al.Periostin reverses high glucose-inhibited osteogenesis of periodontal ligament stem cells via AKT pathway[J].Life Sci,2020,242(12):117184-117196.
[19] LLORENTE I,GARCIA-CASTANEDA N,VALERO C,et al.Osteoporos is in rheumatoid arthritis:dangerous liaisons[J].Front Med (Lausanne),2023,27(11):601618-601629.
[20] YAN Z,WANG Y,SONG Y,et al.Phenethylferulate as a natural inhibitor of inflammation in LPS-stimulated RAW 264.7 macrophages:focus on NF-κB,Akt and Mapk signaling pathways[J].BMC Complement Med Ther,2023,23(1):398-409.
[21] 欧玉琼.阿魏酸对高糖培养MC3T3-E1细胞增殖、分化及炎症因子表达的影响[D].广州：南方医科大学，2023.
[22] KANAZAWA I,SUGIMOTO T J.Diabetes mellitus-induced bone fragility[J].Intern Med,2018,57(19):2773-2785.
[23] SUN Y,ZHU Y,LIU X,et al.Morroniside attenuates high glucose-induced BMSC dysfunction by regulating the Glo1/AGE/RAGE axis.Cell Prolif[J].Cell Prolif,2020,53(8):12866-12881.
基本信息:
中图分类号:R285
引用信息:
[1]张文杰,冯凯,陈意磊,等.高糖环境下药桑提取物脱氧野尻霉素对MC3T3-E1细胞的影响[J].新疆医科大学学报,2024,47(04):471-479.
基金信息:
新疆维吾尔自治区自然科学基金重点项目(2022D01D57); 新疆维吾尔自治区“天山英才”科技领军创新人才项目(2023TSYCLJ0032)
2024-04-15
2024-04-15