持续静压力作用下牙周膜细胞骨保护素、核因子-κB受体活化因子配体表达的增龄性变化

doi:10.7518/hxkq.2022.06.005

华西口腔医学杂志 ›› 2022, Vol. 40 ›› Issue (6): 654-661.doi: 10.7518/hxkq.2022.06.005

持续静压力作用下牙周膜细胞骨保护素、核因子-κB受体活化因子配体表达的增龄性变化

吴洁¹(), 崔占琴¹, 韩瑜², 李文静³()

^1.河北医科大学第二医院口腔正畸科，石家庄 050000
^2.石家庄市人民医院口腔科，石家庄 050011
^3.河北医科大学第二医院口腔内科，石家庄 050000

收稿日期:2022-03-21 修回日期:2022-09-08 出版日期:2022-12-01 发布日期:2022-11-23
通讯作者: 李文静 E-mail:1060324234@qq.com;lwjkq@163.com
作者简介:吴洁，硕士，E-mail：1060324234@qq.com
基金资助:
河北省政府资助临床优秀人才项目(冀财社［2017］46号)

Aging effect of osteoprotegerin and receptor activator of nuclear factor-κB ligand expression in human periodontal ligament cells under continuous static pressure

Wu Jie¹(), Cui Zhanqin¹, Han Yu², Li Wenjing³()

^1.Dept. of Orthodontics, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China
^2.Dept. of Stomatology, Shijiazhuang People’s Hospital, Shijiazhuang 050011, China
^3.Dept. of Oral Medicine, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, China

Received:2022-03-21 Revised:2022-09-08 Online:2022-12-01 Published:2022-11-23
Contact: Li Wenjing E-mail:1060324234@qq.com;lwjkq@163.com
Supported by:
Hebei Provincial Government Funded Clinical Outstanding Talents Project(JCS[2017]46);Correspondence: Li Wenjing, E-mail: lwjkq@163.com

摘要/Abstract

摘要：

目的通过细胞实验观察持续静压力作用下人牙周膜细胞（HPDLCs）表达骨保护素（OPG）、核因子-κB受体活化因子配体（RANKL）的增龄性变化。方法组织块法原代培养HPDLCs，分为A组（13~18岁）、B组（19~29岁）、C组（30~44岁）。CCK-8检测HPDLCs的增殖能力，β-半乳糖苷酶染色检测HPDLCs衰老情况。3组细胞均分别给予0、1.5、3、6、12、24、48、72 h体外持续静压力机械刺激，酶联免疫吸附测定法检测所提上清液中OPG、RANKL的表达。结果体外持续静压力刺激后，牙周膜细胞OPG表达水平随加力时间及年龄增长而增加（P<0.01）。RANKL表达水平随加力时间延长而增加，随年龄增长而下降（P<0.01）。OPG/RANKL比值随加力时间呈先下降后增加趋势，且随年龄增长而增加（P<0.01）。结论体外持续静压力作用下，增龄因素可上调牙周膜细胞OPG表达及OPG/RANKL比值，下调RANKL表达。

关键词: 增龄, 正畸, 人牙周膜细胞, 持续静压力, 骨保护素, 核因子-κB受体活化因子配体

Abstract:

Objective The expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) in human periodontal ligament cells (HPDLCs) was investigated by cell culture under continuous static pressure. Methods HPDLCs were primarily cultured by tissue explant method and divided into three groups: group A (13-18 years old), group B (19-29 years old), and group C (30-44 years old). CCK-8 was used to detect the proliferation of HPDLCs. The senescence of HPDLCs was detected by senescence-associated β-galactosidase staining. Cells in the three groups were respectively given 0, 1.5, 3, 6, 12, 24, 48, and 72 h of continuous static pressure in vitro. The expression of OPG and RANKL in the supernatant was detected by enzyme-linked immunosorbent assay. Results After continuous static pressure in vitro for stimulation, the expression of OPG and RANKL changed. The expression of OPG increased with time and age (P<0.01). The expression of RANKL increased with time and decreased with age (P<0.01). The ratio of OPG/RANKL initially decreased, increased with time, and then continued to rise with age (P<0.01). Conclusion Aging could increase the expression of OPG and the ratio of OPG/RANKL and decrease the expression of RANKL in HPDLCs under continuous static pressure in vitro.

Key words: aging, orthodontics, human periodontal ligament cells, continuous static pressure, osteoprotege-rin, receptor activator of nuclear factor-κB ligand

中图分类号:

R 783.5

吴洁, 崔占琴, 韩瑜, 李文静. 持续静压力作用下牙周膜细胞骨保护素、核因子-κB受体活化因子配体表达的增龄性变化[J]. 华西口腔医学杂志, 2022, 40(6): 654-661.

Wu Jie, Cui Zhanqin, Han Yu, Li Wenjing. Aging effect of osteoprotegerin and receptor activator of nuclear factor-κB ligand expression in human periodontal ligament cells under continuous static pressure[J]. West China Journal of Stomatology, 2022, 40(6): 654-661.

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加力时间/h		空白对照组		压力组
加力时间/h	A组	B组	C组	A组	B组	C组
0	5.572 0±0.347 4	5.845 4±0.445 1	5.521 6±0.196 9	5.571 4±0.179 6	5.393 2±0.339 6	5.414 9±0.584 3
1.5	7.866 4±0.976 6	8.499 1±0.962 6	9.899 7±0.843 3	15.221 4±0.714 3	15.608 5±0.947 9	16.026 6±0.229 2
3	12.771 9±0.754	14.372 8±0.516 0	15.606 0±0.605 0^a	29.084 1±0.930 4	30.479 7±0.455 6	31.461 7±0.496 7
6	20.153 3±1.967 5	23.958 3±1.044 7^a	25.938 6±1.3967 ^a	33.594 7±1.175 3	38.826 7±1.991 5^a	40.096 1±0.740 6^a
12	37.565 4±1.529 6	40.235 7±1.711 2^a	42.015 8±1.996 7^a	89.030 9±1.384 9	92.129 1±1.517 5^a	97.437 3±1.609 0^ab
24	59.576 6±1.804 9	65.217 5±1.443 9^a	71.085 0±1.644 1^ab	118.093 8±1.172 1	124.568 2±3.128 9^a	133.845 6±1.492 3^ab
48	73.786 7±2.926 9	77.833 1±1.850 0^a	80.485 9±1.644 3^ab	162.248 3±3.692 5	169.217 1±2.799 3^a	174.224 2±1.938 0^ab
72	84.806 0±1.875 5	90.283 1±1.679 8^a	96.120 2±1.436 2^ab	224.321 6±1.799 0	229.489 3±1.606 0^a	235.264 5±2.483 3^ab

加力时间/h	空白对照组			压力组
加力时间/h	A组	B组	C组	A组	B组	C组
0	0.272 8±0.016 9^cdefg	0.266 4±0.016 9^cdefg	0.284 1±0.016 9^defd	0.278 3±0.015 2	0.239 6±0.011 6	0.233 6±0.016 9
1.5	0.340 4±0.026 6^cdefg	0.314 7±0.044 2^cdefg	0.306 8±0.066 3^defg	0.948 5±0.011 7	0.870 8±0.073 0	0.647 8±0.187 9
3	0.533 8±0.059 7^defg	0.493 1±0.024 0^defg	0.458 8±0.053 1^defg	2.580 0±0.099 4	1.867 6±0.175 5	1.503 2±0.159 6^a
6	0.908 6±0.159 6	0.848 2±0.226 2	0.775 5±0.257 1	4.749 3±0.138 3	3.396 0±0.787 2^a	2.857 8±0.515 2^a
12	1.6608±0.267 0^cef	1.350 8±0.254 4^ef	1.195 9±0.242 9	6.893 3±0.754 3	5.515 7±0.891 5^a	5.052 6±0.701 2^a
24	2.620 5±0.597 2^cdf	2.482 7±0.664 0^cdf	2.140 1±0.337 1^cd	8.889 2±0.714 8	6.458 1±0.402 6^a	5.390 3±0.992 6^ab
48	3.109 3±0.434 9^cd	2.759 4±0.334 9^cd	2.595 9±0.223 5^acd	10.003 4±0.864 6	7.546 9±0.854 6^a	6.185 1±0.376 6^ab
72	3.425 3±0.342 4^cde	3.289 7±0.611 0^cdef	2.952 0±0.453 0^cde	11.602 8±0.768 5	8.386 9±0.291 8^a	7.182 3±0.722 1^ab

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持续静压力作用下牙周膜细胞骨保护素、核因子-κB受体活化因子配体表达的增龄性变化

Aging effect of osteoprotegerin and receptor activator of nuclear factor-κB ligand expression in human periodontal ligament cells under continuous static pressure

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