文献类型：期刊文献

中文题名：PI3K/Akt信号通路在磷酸三钙磨损颗粒诱导小鼠颅骨溶解中的作用

英文题名：Role of PI3K/Akt Signaling Pathway in the TCP Wear Particles-induced Calvarial Osteolysis in Mice Model

作者：张雨笛[1];严明[2];俞立虹[1];章思烨[1];诸葛建颖[1];郭健恒[1];丁樑[1];张云[1]

机构：[1]绍兴文理学院医学院;[2]杭州电子科技大学生命信息与仪器工程学院

年份：2017

卷号：36

期号：3

起止页码：212

中文期刊名：中国运动医学杂志

外文期刊名：Chinese Journal of Sports Medicine

收录：CSTPCD、、北大核心2014、CSCD2017_2018、北大核心、CSCD

基金：浙江省自然科学基金(No.LY17H060007;No.LY15H180012)

语种：中文

中文关键词：磷酸三钙(TCP)磨损颗粒;骨溶解;破骨细胞;PI3K/Akt信号通路

外文关键词：TCP wear particles; osteolysis; osteoclasts; PI3K/Akt signaling pathway

中文摘要：目的:研究PI3K/Akt信号通路在磷酸三钙(tricalcium phosphate,TCP)磨损颗粒诱导小鼠颅骨溶解中的作用。方法:36只雄性ICR小鼠随机分为假手术组(Sham)、模型组(TCP)和LY294002处理组,每组12只。取TCP磨损颗粒30 mg置于颅顶后缝合皮肤构建小鼠颅骨溶解模型。LY294002处理组小鼠于术后第2天颅顶局部注射PI3K抑制剂LY294002(5 mg·kg^(-1)),每周3次;假手术组小鼠颅骨顶仅注射生理盐水,注射时间与LY294002一致。2周后处死动物取骨膜和颅骨。通过Micro-CT分析小鼠颅骨溶解情况、骨密度(bone mineral density,BMD)和骨矿含量(bone mineral content,BMC);HE染色观察颅骨表面炎症反应及破骨细胞(osteoclasts)形成;Real-time PCR检测骨组织中破骨细胞生成标志物抗酒石酸酸性磷酸酶(tartrate-resistant acid phosphatase,TRAP)、组织蛋白酶K(cathepsin K,Cst K)、核因子κB受体活化因子配体(receptor activator for nuclear factor-κB kigand,RANKL)和c-Fos的m RNA水平;ELISA检测骨膜中炎症因子TNF-α、IL-1β和IL-6等水平;Western Blotting检测颅骨组织Akt、p-Akt Ser473和p-AktThr308等蛋白表达变化。结果:Micro-CT和组织形态学分析结果显示,PI3K抑制剂LY294002能明显抑制TCP磨损颗粒诱导小鼠颅骨溶解和破骨细胞形成(P<0.05),增加BMD和BMC含量(P<0.05);下调破骨细胞生成标志物TRAP、Cst K、RANKL和c-Fos等m RNA水平(P<0.05),并抑制炎症因子TNF-α、IL-1β和IL-6的释放(P<0.05);而且LY294002显著减弱TCP磨损颗粒诱导的Akt信号蛋白活化,导致p-Akt S-er473和p-AktThr308等蛋白表达明显下调(P<0.05)。结论:PI3K/Akt信号通路参与调控TCP磨损颗粒诱导的假体周围骨溶解,可作为假体周围骨溶解和关节松动的治疗靶标。

外文摘要：Objective To explore the role of the PI3K/Akt signaling pathway in the calvarial osteoly-sis induced by TCP wear particles in mice model. Methods Thirty-six male ICR mice were randomlydivided into a sham group（n=12）,TCP group（n=12）and a LY294002-treated group（n=12）. A mu-rine calvarial model of osteolysis was established through implanting 30 mg of TCP particles onto thesurface of bilateral parietal bones following the removal of the periosteum. On the second postoperativeday,LY294002（5 mg·kg^（-1））was locally injected to the calvarium under the periosteum three times aweek;mice in the sham group received local injection of normal saline（N.S.）in the calvarium,and the injection time was consistent with that of LY294002. Two weeks later,the calvaria and periostea wereobtained after the mice were executed. The calvarial osteolysis,bone mineral density（BMD）and bonemineral content（BMC）were analyzed using Micro-CT,Hematoxylin-Eosin（HE）staining was conductedto observe the inflammatrory response and formation of osteoclasts. Real-time PCR was applied to de-tect the m RNA level of tartrate-resistant acid phosphatase（TRAP）,the marker of osteoclasts forma-tion,cathepsin K（Cst K）,receptor activator for nuclear factor-κB kigand（RANKL）and c-Fos. The re-lease of tumor necrosis factor-α（TNF-α）,interleukin-6（IL-6）and IL-1β were measured using en-zyme-linked immumsorbent assay（ELISA）. Results Micro-CT and histological analysis indicated thatLY294002,the specific inhibitor of PI3 K,significantly prevented TCP wear particles-induced osteolysisand osteoclastogenesis,and increased BMD and BMC in the calvaria of mice. Real-time PCR data re-vealed LY294002 significantly suppressed the increase in m RNA level of osteoclastogenic genes suchas TRAP,Cst K,RANKL and c-Fos in the calvaria of TCP wear particles-implanted group. ELISA as-say showed that TCP wear particles-induced release of TNF-α,IL-1β and IL-6 was significantly inhib-ited by LY294002 treatment. Furthermore,LY294002 significantly attenuated TCP wear particles-trig-gered activation of Akt,and down-regulated the level of p-Akt Ser473 and p-AktThr308. Conclusion PI3K/Akt signaling pathway contributes to TCP wear particle-induced osteolysis,and can be developedas a new therapeutic target for the prevention and treatment of bone destruction diseases caused bywear debris.