抑制miR-145表达影响小鼠牙种植体周围炎炎症反应的实验研究

Inhibition of miR-145 expression affects the inflammatory response in experimental peri-implantitis in mice

目的:探讨抑制miR-145对小鼠种植体周围炎炎症反应的影响。方法:将C57BL/6J雄性小鼠40只随机分为正常对照组、炎症对照组、炎症miR-NC组、炎症miR-inhibitor组。在各组上颌植入种植体,对3个炎症组拴丝形成种植体周围炎后,对炎症miR-inhibitor组局部注射miR-145抑制剂,在炎症miR-NC组局部注射miR-145抑制剂的阴性对照剂,微计算机断层扫描观察各组种植体周围骨丧失情况,苏木素-伊红染色观察种植体周围组织炎细胞浸润情况,实时荧光定量PCR检测种植体周围牙龈组织miR-145、趋化因子CC基序配体2(CCL2)、肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素10(IL-10)、基质金属蛋白酶8(MMP8)的表达水平。结果:与正常对照组小鼠相比,3个炎症组小鼠骨丧失量及炎细胞浸润明显增加(P<0.05),牙龈组织中炎症相关因子CCL2、TNF-α、IL-1β、IL-10、MMP8表达水平升高(P<0.05);抑制miR-145表达后,种植体周围炎小鼠骨丧失量、炎细胞浸润情况改善,CCL2、TNF-α、IL-1β、MMP8表达减少,IL-10表达升高(P<0.05)。结论:抑制miR-145的表达可缓解种植体周围炎炎症反应,可能与其调节牙龈组织中炎症相关因子表达有关。

Objective:To investigate the effect of inhibiting miR-145 on inflammatory response in experimental peri-implantitis in mice.Methods:Forty male C57BL/6J mice were randomly divided into four groups:normal control group,peri-implantitis control group,peri-implantitis miR-NC group,and peri-implantitis miR-inhibitor group.All groups received dental implant insertion in maxillae,then the three peri-implantitis groups received ligation around implants to induce peri-implantitis.Afterwards,miR-145 inhibitor was injected into peri-implant gingiva in peri-implantitis miR-inhibitor group,and its negative control reagent was injected in peri-implantitis miR-NC group.Peri-implant bone loss in each group was detected using micro-computer tomography,and infiltration of inflammatory cells around the implants was examined using hematoxylin and eosin staining.Expression levels of miR-145,C-C motif chemokine ligand 2(CCL2),tumor necrosis factorα(TNF-α),interleukin 1β(IL-1β),interleukin 10(IL-10)and matrix metalloproteinase 8(MMP8)in peri-implant gingival tissues were detected using real-time quantitative PCR.Results:Compared with normal control group,bone loss and inflammatory cell infiltration in all peri-implantitis groups were greater(P<0.05),and the expressions of inflammation-related factors CCL2,TNF-α,IL-1β,IL-10 and MMP8 were increased(P<0.05).Upon miR-145 inhibition,the bone loss and inflammatory cell infiltration were ameliorated,and the expression of inflammation-related factors CCL2,TNF-α,IL-1βand MMP8 was decreased,while the expression of IL-10 enhanced(P<0.05).Conclusions:Inhibition of miR-145 expression could suppress peri-implantitis,which may be associated with the regulationof inflammation-related factor expression in peri-implant gingival tissues.