脱蛋白牛骨基质对骨膜细胞影响巨噬细胞M1表型的调控  

Deproteinized bovine bone matrix regulates the effect of periosteal cells on macrophage M1 phenotype

作　　者：黄翀 陈斯 陈冬[1] 夏海斌[1] HUANG Chong;CHEN Si;CHEN Dong;XIA Hai-bin(School of Stomatology,Wuhan University,Hubei Wuhan 430079)

机构地区：[1]武汉大学口腔医学院,湖北武汉430079

出　　处：《临床口腔医学杂志》2021年第3期141-144,共4页Journal of Clinical Stomatology

基　　金：国家自然科学基金资助项目(82071095,31570982)。

摘　　要：目的:探讨脱蛋白牛骨基质骨替代材料与炎症环境中骨膜细胞的条件培养液对巨噬细胞M1表型的影响。方法:取P3代小鼠颅骨骨膜细胞,10 ng/mL LPS刺激24 h,分别换成常规培养基和脱蛋白牛骨基质(Bio-Oss骨粉)浸提液培养24 h,取上清记为A液、B液。体外培养小鼠RAW264.7细胞,100 ng/mL LPS诱导成为M1型巨噬细胞,分别加入条件培养液A液(对照组)和B液(实验组)。培养24 h后CCK-8检测细胞增殖活性;qRT-PCR分析M1型表型基因诱导性一氧化氮合酶(inducible nitric oxide synthase,iNOS)、肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)。免疫荧光检测iNOS的表达。结果:CCK-8结果显示,实验组M1巨噬细胞增殖活性提高。与对照组相比,实验组iNOS、TNF-α、IL-1β的mRNA表达均下调(P<0.05);免疫荧光结果显示,实验组的iNOS表达下调。结论:脱蛋白牛骨基质可抑制骨膜细胞间接作用下的巨噬细胞M1表型基因的表达。Objective:To investigate the effect of deproteinized bovine bone matrix which is one of bone replacement material and conditioned culture medium of inflammatory periosteal cells on M1 phenotype macrophage.Methods:P3 generation mouse skull periosteum cells were taken and stimulated with 10 ng/mL LPS for 24 h,then replaced with conventional medium and deproteinized bovine bone matrix(Bio-Oss)extract medium for 24 h,respectively.The supernatant was recorded as liquid A and liquid B.Mouse RAW264.7 cells were cultured in vitro and induced into M1-type macrophages by 100 ng/mL LPS.The above-mentioned conditional culture medium was added respectively,the control group was added with liquid A and experimental group with liquid B.CCK-8 was used to measure cell proliferation activity after 24 h culture.qRT-PCR was used to analyze the inducible nitric oxide synthase(iNOS),tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)of the M1-type phenotype genes.The expression of iNOS was detected by immunofluorescence.Results:CCK-8 results showed that the proliferation activity of M1 macrophages in experimental group improved.The mRNA expressions of iNOS,TNF-αand IL-1βin the experimental group were down-regulated compared with those in the control group(P<0.05).The iNOS expression was also down-regulated in experimental group showed by immunofluorescence.Conclusion:Deproteinized bovine bone matrix can affect periosteal cells in inhibition of the expression of M1 phenotype gene in macrophages.

关 键 词：骨膜细胞 巨噬细胞 脱蛋白牛骨基质 

分 类 号：Q813.1[生物学—生物工程]