Effect of estrogen deficiency on the expression levels of nuclear factor-kappa B and interleukin-17 in periodontal tissues

doi:10.3969/j.issn.2095-4344.2017.24.005

Abstract

Abstract:

BACKGROUND: There is a very close relationship between osteoporosis and periodontal disease in postmenopausal women, but the mechanism remains unclear.
OBJECTIVE: To investigate the expression of nuclear factor-κB in the alveolar bone and interleukin-17 in the serum and gingiva in the mouse model of osteoporosis caused by ovariectomy.
METHODS: Female mice aged 3 months were randomly divided into ovariectomy and sham operation groups. At 6 months after surgery, the mouse models were evaluated histologically on the submandibular bone and thigh bone stained with hematoxylin and eosin. In the submandibular bone, the expression levels of OCN and Runx2 were detected by RT-PCR, and the expression level of nuclear factor-κB was detected by immunohistochemical staining and western blot assay. Besides, the expression level of interleukin-17 in the serum and gingival homogenate was evaluated using Cytometric Beads Array.
RESULTS AND CONCLUSION: The thigh bone in the ovariectomy group revealed the thin cortical bone, enlarged marrow cavity, and increased resorption lacunae, as well as fewer, thinner trabeculae with lower density and irregular structure. Compared with the sham operation group, the expression levels of OCN and Runx2 in the alveolar bone were decreased in the ovariectomy group. The activation of nuclear factor-κB (P65) appeared with P65 positive expression in the submandibular bone in the ovariectomy group, and the relative expression level was higher than that in the sham operation group. The serum level of interleukin-17 in the ovariectomy group was higher than that in the sham operation group, but the level in the gingival tissue showed no significant difference between the two groups. These results indicate that estrogen deficiency after ovariectomy can activate nuclear factor-κB signal pathway to play a role in periodontal osteolysis. However interleukin-17 in the local periodontal tissue may not be a key cytokine to damage the periodontal tissue.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Osteoporosis, Postmenopausal, Ovariectomy, NF-kappa B, Interleukin-17, Tissue Engineering

CLC Number:

R318

Jia Zhi, Zhang Feng-wei, Qi Xing-ying, Yan Xiao, Hua Ye, Zhao Meng-ming, Liu Da-yong. Effect of estrogen deficiency on the expression levels of nuclear factor-kappa B and interleukin-17 in periodontal tissues[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(24): 3796-3802.

Figures/Tables 1

2.1 雌激素缺乏状态下股骨及下颌骨组织学变化苏木精-伊红染色结果显示：假手术组小鼠股骨骨小梁密集排列，互相连接成网状结构，骨小梁厚度大，其间距小(图1A)；卵巢切除小鼠骨小梁连接中断，明显稀少，明显变细，间距增宽(图1B)，牙槽骨高度降低，牙槽骨吸收；骨小梁的间隙较大，骨组织界面可以看见较多的骨吸收陷窝及破骨细胞，提示骨代谢呈骨吸收活跃状态，箭头所示分别为水平型吸收的牙槽骨和骨陷窝(图2)。 2.2 雌激素缺乏状态下成骨相关因子表达变化对小鼠下颌骨骨组织总RNA进行紫外分光光度计检测，A260/A280 nm大于1.8，提示RNA纯度较好；RNA样品经琼脂糖凝胶电泳，可清晰看见28 s和18 s条带，提示RNA未降解(图3A)。电泳所得成骨相关因子OCN和Runx2的RT-PCR凝胶图像见图3B。经Quantity one进行灰度值分析，卵巢切除组下颌骨OCN相对表达量为0.166 8±0.047 0，假手术组为0.539 1±0.129 3，差异有显著性意义(P < 0.05)；卵巢切除组下颌骨Runx2相对表达量为0.306 4±0.048 3，假手术组为0.581 7±0.113 1，差异有显著性意义(P < 0.05)。 2.3 雌激素缺乏状态下NF-κB表达变化免疫组化染色结果显示：卵巢切除组小鼠下颌骨中软骨细胞和成骨细胞NF-κB P65均呈阳性表达，而假手术组未见明显NF-κB P65表达(图4，5)。 Western-blot结果显示：卵巢切除组小鼠牙槽骨NF-κB P65的条带较假手术组条带颜色清晰且粗大(图6A)；成像扫描经Quantity one进行灰度值分析，卵巢切除组下颌骨NF-κB P65相对表达量为0.585 4±0.043 0，假手术组为0.104 5±0.027 0，差异有显著性意义(P < 0.05，图6B)。 2.4 雌激素缺乏状态下白细胞介素17表达变化卵巢切除组小鼠血清中白细胞介素17相对表达量为3.313± 0.203，假手术组为1.417±0.204，差异有显著性意义(P < 0.05)；而在牙龈组织中，卵巢切除组小鼠白细胞介素17相对表达量为1.050±0.703，假手术组为1.167±0.662，差异无显著性意义(P > 0.05)。"

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