Page 35 - 《中国药房》2022年19期

模拟微重力下柚皮苷对T细胞共育的成骨细胞增殖分化的影响 Δ

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陈建兴 ，尹文哲 ，孙奇峰，周 森，张亚龙，王 翌（哈尔滨医科大学附属第二医院骨四科，哈尔滨 150086）
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中图分类号 R965 文献标志码 A 文章编号 1001-0408（2022）19-2333-05
DOI 10.6039/j.issn.1001-0408.2022.19.06

摘 要 目的 在模拟微重力情况下，探索柚皮苷作用于T细胞-成骨细胞共育体系后，对成骨细胞增殖分化的影响。方法 本实
验设正常重力对照组、正常重力柚皮苷组、模拟微重力对照组、模拟微重力柚皮苷组，其中对照组分别在正常重力或模拟微重力条
件下将T细胞与成骨细胞共育，柚皮苷组分别在对照组基础上加入1×10 mol/L柚皮苷溶液。显微镜下观察成骨细胞形态学，
－5
CCK-8法检测成骨细胞增殖率，碱性磷酸酶（ALP）试剂盒检测ALP活性，实时荧光定量聚合酶链式反应检测Runt相关转录因子2
（Runx2）、白细胞介素6（IL-6）mRNA相对表达水平，Western blot法检测Runx2和IL-6蛋白相对表达水平。结果 与正常重力对照
组比较，模拟微重力对照组成骨细胞密度降低，仅有少量聚集，细胞形态以圆形居多；与模拟微重力对照组比较，模拟微重力柚皮
苷组成骨细胞成梭形或多角形，形态饱满，成鱼群状聚集。与正常重力对照组比较，正常重力柚皮苷组成骨细胞增殖率、ALP活
性、Runx2 mRNA和蛋白相对表达水平显著升高，IL-6 mRNA和蛋白相对表达水平显著降低（P＜0.05）；模拟微重力对照组成骨细
胞增殖率、ALP活性、Runx2 mRNA和蛋白相对表达水平显著降低，IL-6 mRNA和蛋白相对表达水平显著升高（P＜0.05）。与模拟
微重力对照组比较，模拟微重力柚皮苷组成骨细胞增殖率、ALP活性、Runx2 mRNA和蛋白相对表达水平显著升高，IL-6 mRNA和
蛋白相对表达水平显著降低（P＜0.05）。结论 在T细胞-成骨细胞共育体系中，模拟微重力可以抑制成骨细胞的增殖分化能力；该
条件下柚皮苷可以改善成骨细胞的增殖分化能力，其作用机制与IL-6水平的降低及Runx2水平的升高有关。
关键词 柚皮苷；成骨细胞；模拟微重力；增殖分化；T细胞

Effects of naringin on the proliferation and differentiation of osteoblasts co-cultured with T cells under
simulated microgravity
CHEN Jianxing，YIN Wenzhe，SUN Qifeng，ZHOU Sen，ZHANG Yalong，WANG Yi（Dept. Four of Orthopaedics，
the Second Affiliated Hospital of Harbin Medical University，Harbin 150086，China）

ABSTRACT OBJECTIVE To explore the effects of naringin on the proliferation and differentiation of osteoblasts in T cell-
osteoblast co-culture system under simulated microgravity. METHODS This experiment was setting normal gravity control group，
normal gravity naringin group，simulated microgravity control group and simulated microgravity naringin group. In the control
group，T cells were co-cultured with osteoblasts under normal gravity or simulated microgravity conditions. In the naringin group，
1×10 －5 mol/L naringin solution was added additionally on the basis of control group.The morphology of osteoblasts was observed
under the microscope. The proliferation rate of osteoblasts was detected by CCK-8 method. The alkaline phosphatase（ALP）activity
was detected by ALP kit. mRNA relative expressions of Runt related transcription factor 2（Runx2）and interleukin-6（IL-6）were
detected by real-time fluorescence quantitative polymerase chain reaction. The relative expression of Runx2 and IL-6 protein were
detected by Western blot. RESULTS Compared with normal gravity control group，the density of osteoblasts in the simulated
microgravity control group decreased，only a small number of cells were aggregated，and the cell morphology was mostly round；
compared with the simulated microgravity control group，the osteoblasts in the simulated microgravity naringin group formed
spindle or polygonal，plump in shape，and clustered in groups of fish. Compared with normal gravity control group，the
proliferation rate of osteoblasts，ALP activity，relative expressions of Runx2 mRNA and protein in the normal gravity naringin
group were significantly increased，while the relative expressions of IL-6 mRNA and protein were significantly decreased（P＜
0.05）；the proliferation rate of osteoblasts，ALP activity，relative expressions of Runx2 mRNA and protein in the simulated
microgravity control group were significantly decreased，while relative expressions of IL-6 mRNA and protein were significantly
increased（P＜0.05）. Compared with the simulated microgravity control group，the proliferation rate of osteoblasts，ALP activity，
relative expressions of Runx2 mRNA and protein in the simulated microgravity naringin group were significantly increased，while
relative expressions of IL-6 mRNA and protein were significantly decreased（P＜0.05）. CONCLUSIONS In the T cell-osteoblast
co-culture system，simulated microgravity can inhibit the proliferation and differentiation of osteoblasts；naringin can improve the
proliferation and differentiation ability of osteoblasts under this
Δ 基金项目 国家自然科学基金资助项目（No.81873093）
condition， and its mechanism of action is related to the
＊ 第一作者 硕 士 研 究 生 。 研 究 方 向 ：中 药 基 础 。 E-mail：
jianxingchen2021@126.com decrease of IL-6 level and the increase of Runx2 level.
# 通信作者 教授，博士生导师。研究方向：骨质疏松及关节疾 KEYWORDS naringin；osteoblasts；simulated microgravity；
病。电话：0451-86297531。E-mail：yinwenzhe@medmail.com.cn proliferation and differentiation；T cell

中国药房 2022年第33卷第19期 China Pharmacy 2022 Vol. 33 No. 19 ·2333·