中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Trauma
2015年
9期
850-855
,共6页
邵洪国%崔京福%朱世军%王熠军%朱锋%杨惠林%耿德春%徐耀增
邵洪國%崔京福%硃世軍%王熠軍%硃鋒%楊惠林%耿德春%徐耀增
소홍국%최경복%주세군%왕습군%주봉%양혜림%경덕춘%서요증
关节成形术,置换%骨质溶解%破骨细胞%钛
關節成形術,置換%骨質溶解%破骨細胞%鈦
관절성형술,치환%골질용해%파골세포%태
Arthroplasty,replacement%Osteolysis%Osteoclasts%Titanium
目的 探讨淫羊藿苷对钛颗粒诱导炎症性骨溶解的治疗作用. 方法 雄性C57BL/J6小鼠84只,按随机数字表法分为空白组、对照组、淫羊藿苷低剂量治疗组(L组)和高剂量治疗组(H组),每组21只.治疗组采用钛颗粒诱导小鼠颅骨溶解模型,并给予不同浓度的淫羊藿苷:L组0.1 mg·g-1·d-1,H组0.3mg·g-1 ·d-1,持续至建模后14 d.采用显微CT评估骨溶解程度;HE染色观察细胞浸润程度;形态计量学测量骨溶解面积(EBS);抗酒石酸酸性磷酸酶(TRAP)染色检测成熟破骨细胞;定量RT-PCR检测降钙素受体(CTR)、TRAP、组织蛋白酶K(Cath-K)和激活T细胞的核因子c1(NFATc1)基因mRNA含量;免疫组织化学检测肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和IL-6的表达变化;酶联免疫吸附(ELISA)法检测核因子-κB受体活化因子配体(RANKL)和骨保护素(OPG)的表达量. 结果 显微CT显示治疗组颅骨骨溶解现象明显减轻,骨密度、骨体积分数L组和H组分别为(89.44±18.51) mg/cm2、(48.32±9.54)％和(144.85±19.49) mg/cm2、(65.67 ±6.80)％,与对照组(61.23±13.41) mg/cm2、(27.83±7.51)％比较,差异有统计学意义(P＜0.05).HE染色表明,对照组骨膜内有大量巨噬样细胞,L组和H组炎症反应明显减轻.对照组EBS为(0.14±O.03) mm2,L组和H组EBS降为(0.07 ±0.01)mm2和(0.04 ±0.01) mm2 (P ＜0.05).对照组颅骨溶解侧可见大量TRAP阳性细胞,治疗后TRAP阳性细胞明显减少.免疫组织化学染色表明淫羊藿苷能抑制TNF-α、IL-1β和IL-6的表达.RT-PCR显示,钛颗粒上调CTR、TRAP、Cath-K和NFATc1基因mRNA表达,经治疗后上述基因mRNA表达水平较低,与对照组比较差异有统计学意义(P＜0.05).ELISA法检测结果显示,对照组RANKL和OPG的表达量分别为(23.02 ±4.39) pg/ml和(1 360.52 ±514.08) pg/ml,与治疗组[L组:(16.87±4.75) pg/ml、(1 890.54±550.94) pg/ml,H组:(12.80±5.77) pg/ml、(2 001.67±325.01) pg/ml]比较,差异有统计学意义(P＜0.05). 结论 淫羊藿苷能降低磨损颗粒引起的炎症反应,抑制破骨细胞活化,减轻骨溶解.
目的 探討淫羊藿苷對鈦顆粒誘導炎癥性骨溶解的治療作用. 方法 雄性C57BL/J6小鼠84隻,按隨機數字錶法分為空白組、對照組、淫羊藿苷低劑量治療組(L組)和高劑量治療組(H組),每組21隻.治療組採用鈦顆粒誘導小鼠顱骨溶解模型,併給予不同濃度的淫羊藿苷:L組0.1 mg·g-1·d-1,H組0.3mg·g-1 ·d-1,持續至建模後14 d.採用顯微CT評估骨溶解程度;HE染色觀察細胞浸潤程度;形態計量學測量骨溶解麵積(EBS);抗酒石痠痠性燐痠酶(TRAP)染色檢測成熟破骨細胞;定量RT-PCR檢測降鈣素受體(CTR)、TRAP、組織蛋白酶K(Cath-K)和激活T細胞的覈因子c1(NFATc1)基因mRNA含量;免疫組織化學檢測腫瘤壞死因子-α(TNF-α)、白細胞介素-1β(IL-1β)和IL-6的錶達變化;酶聯免疫吸附(ELISA)法檢測覈因子-κB受體活化因子配體(RANKL)和骨保護素(OPG)的錶達量. 結果 顯微CT顯示治療組顱骨骨溶解現象明顯減輕,骨密度、骨體積分數L組和H組分彆為(89.44±18.51) mg/cm2、(48.32±9.54)％和(144.85±19.49) mg/cm2、(65.67 ±6.80)％,與對照組(61.23±13.41) mg/cm2、(27.83±7.51)％比較,差異有統計學意義(P＜0.05).HE染色錶明,對照組骨膜內有大量巨噬樣細胞,L組和H組炎癥反應明顯減輕.對照組EBS為(0.14±O.03) mm2,L組和H組EBS降為(0.07 ±0.01)mm2和(0.04 ±0.01) mm2 (P ＜0.05).對照組顱骨溶解側可見大量TRAP暘性細胞,治療後TRAP暘性細胞明顯減少.免疫組織化學染色錶明淫羊藿苷能抑製TNF-α、IL-1β和IL-6的錶達.RT-PCR顯示,鈦顆粒上調CTR、TRAP、Cath-K和NFATc1基因mRNA錶達,經治療後上述基因mRNA錶達水平較低,與對照組比較差異有統計學意義(P＜0.05).ELISA法檢測結果顯示,對照組RANKL和OPG的錶達量分彆為(23.02 ±4.39) pg/ml和(1 360.52 ±514.08) pg/ml,與治療組[L組:(16.87±4.75) pg/ml、(1 890.54±550.94) pg/ml,H組:(12.80±5.77) pg/ml、(2 001.67±325.01) pg/ml]比較,差異有統計學意義(P＜0.05). 結論 淫羊藿苷能降低磨損顆粒引起的炎癥反應,抑製破骨細胞活化,減輕骨溶解.
목적 탐토음양곽감대태과립유도염증성골용해적치료작용. 방법 웅성C57BL/J6소서84지,안수궤수자표법분위공백조、대조조、음양곽감저제량치료조(L조)화고제량치료조(H조),매조21지.치료조채용태과립유도소서로골용해모형,병급여불동농도적음양곽감:L조0.1 mg·g-1·d-1,H조0.3mg·g-1 ·d-1,지속지건모후14 d.채용현미CT평고골용해정도;HE염색관찰세포침윤정도;형태계량학측량골용해면적(EBS);항주석산산성린산매(TRAP)염색검측성숙파골세포;정량RT-PCR검측강개소수체(CTR)、TRAP、조직단백매K(Cath-K)화격활T세포적핵인자c1(NFATc1)기인mRNA함량;면역조직화학검측종류배사인자-α(TNF-α)、백세포개소-1β(IL-1β)화IL-6적표체변화;매련면역흡부(ELISA)법검측핵인자-κB수체활화인자배체(RANKL)화골보호소(OPG)적표체량. 결과 현미CT현시치료조로골골용해현상명현감경,골밀도、골체적분수L조화H조분별위(89.44±18.51) mg/cm2、(48.32±9.54)％화(144.85±19.49) mg/cm2、(65.67 ±6.80)％,여대조조(61.23±13.41) mg/cm2、(27.83±7.51)％비교,차이유통계학의의(P＜0.05).HE염색표명,대조조골막내유대량거서양세포,L조화H조염증반응명현감경.대조조EBS위(0.14±O.03) mm2,L조화H조EBS강위(0.07 ±0.01)mm2화(0.04 ±0.01) mm2 (P ＜0.05).대조조로골용해측가견대량TRAP양성세포,치료후TRAP양성세포명현감소.면역조직화학염색표명음양곽감능억제TNF-α、IL-1β화IL-6적표체.RT-PCR현시,태과립상조CTR、TRAP、Cath-K화NFATc1기인mRNA표체,경치료후상술기인mRNA표체수평교저,여대조조비교차이유통계학의의(P＜0.05).ELISA법검측결과현시,대조조RANKL화OPG적표체량분별위(23.02 ±4.39) pg/ml화(1 360.52 ±514.08) pg/ml,여치료조[L조:(16.87±4.75) pg/ml、(1 890.54±550.94) pg/ml,H조:(12.80±5.77) pg/ml、(2 001.67±325.01) pg/ml]비교,차이유통계학의의(P＜0.05). 결론 음양곽감능강저마손과립인기적염증반응,억제파골세포활화,감경골용해.
Objective To determine the therapeutic effect of icariin in the treatment of titamum particle-induced inflammatory osteolysis and examine the relationship between icariin and receptor activator of NF-κB ligand (RANKL)/osteoprotegerin (OPG).Methods Eighty-four male C57BL/J6 mice were divided into four groups (21 animals for each) according to the random number table:black group,control group (titanium particles only),low-dosage icariin group (0.1 mg · g-1 · d-1,L group),and high-dosage icariin group (0.3 mg · g-1 · d-1,H group).The animal model of titanium particleinduced calvaria osteolysis was used in this study.After 2 weeks mice calvariae were harvested for microCT,histological and molecular analysis.Results As assessed by the micro-CT,icariin reduced particle-induced osteolysis:bone mineral density and bone volume fraction in L group was (89.44 ± 18.51) mg/cm2 and (48.32 ± 9.54) ％ and was (1 44.85 ± 19.49) mg/cm2 and (65.67 ± 6.80) ％ in H group as compared to (61.23 ± 13.41) mg/cm2 and (27.83 ± 7.51) ％ in control group (P ＜ 0.05).HE staining demonstrated diminished inflammation after treatment with icariin while intensive macrophage infiltration into the calvariae in control group.Histomorphometric results demonstrated the area of eroded bone surface (EBS) was reduced to (0.07 ± 0.01) mm2 and (0.04 ± 0.01) mm2 respectively in L and H groups compared to (0.14 ± 0.03) mm2 in control group (P ＜ 0.05).Tartrate resistant acid phosphatase (TRAP) staining revealed multiple TRAP-positive cells accumulated along the EBS in control group,whereas the number of TRAP-positive cells was reduced by icariin.Immunohistological examination showed titanium particle up-regulated expressions of tumor necrosis factor-α (TNF-α),interleukin (IL)-1β and IL-6.In contrast,low expressions were noted after icariin treatment.RT-PCR showed titanium particle stimulation could enhance mRNA expressions of calcitonin receptor (CTR),TRAP,cathepsin-K (Cath-K) and nuclear factor of activated T-cells,cytoplasmic 1 (NFATc1) in the clavariae tissues.However,the mRNA levels of these genes were markedly reduced after icariin treatment.ELISA results showed levels of RANKL and OPG were (23.02 ± 4.39) pg/ml and (1 360.52 ± 514.08) pg/ml in control group,but secretion of RANKL was reduced together with an increase in OPG by icariin [L group:(16.87 ± 4.75),(1 890.54 ± 550.94) pg/ml;H group:(12.80 ± 5.77),(2 001.67 ± 325.01)pg/ml,P ＜ 0.05].Conclusion Icariin can effectively inhibit titanium particle-induced inflammatory osteolysis by suppressing osteoclast activity and attenuating osteolysis.
