伊班膦酸钠对膝骨关节炎大鼠自噬相关蛋白LC3 Ⅱ的影响  

The effect of ibandronte sodium on autophagy-related protein LC3 Ⅱ in rat model of knee osteoarthritis

作　　者：周桂娟[1] 周君[1,2,3] 肖子建 孙光华 伍琦[1] 曾亚华[1] 刘静[1] 钟培瑞 成果 王甜甜 邓程远 廖瑛 ZHOU Gui-Juan;ZHOU Jun;XIAO Zi-Jian;SUN Guang-Hua;WU Qi;ZENG Ya-Hua;LIU Jing;ZHONG Pei-Rui;CHENG Guo;WANG Tian-Tian;DENG Cheng-Yuan;LIAO Ying(Department of Rehabilitation,The First Affiliated Hospital of University of South China,Hengyang 421001,China;Department of Rehabilitation,West China Hospital,Sichuan University,Chengdu 610041,China;Key Laboratory of Rehabilitation Medicine in Sichuan,Chengdu 610041,China)

机构地区：[1]南华大学附属第一医院康复医学科,衡阳421001 [2]四川大学华西医院康复医学中心,成都610041 [3]康复医学四川省重点实验室,成都610041

出　　处：《中国疼痛医学杂志》2020年第2期91-97,共7页Chinese Journal of Pain Medicine

基　　金：国家自然科学基金面上项目(81674045);国家自然科学基金青年项目(81804155);湖南省卫生计生委科研计划课题项目 (C20180143)。

摘　　要：目的:研究伊班膦酸钠对膝骨关节炎大鼠自噬相关蛋白LC3 Ⅱ的表达变化,初步探讨软骨组织中自噬相关蛋白水平与骨关节炎(osteoarthritis, OA)发生发展的关系。方法:将30只3月龄雌性SD大鼠随机分为3组,每组10只:假手术组(Sham)、生理盐水治疗组(ACLT+NS)和伊班膦酸钠治疗组(ACLT+IB)。ACLT+NS组、ACLT+IB组行膝关节前交叉韧带切断术(ACLT)建立膝骨关节炎模型。术后1周ACLT+IB组予以伊班膦酸钠干预。检测指标:番红染色、Mankin’s评分;关节软骨下骨形态学检测;大鼠血I型胶原C端肽(CTX-I)、Ⅱ型胶原C端肽(CTX-Ⅱ)及IL-1、IL-6水平;基质金属蛋白酶-13(MMP-13)及自噬相关蛋白LC3 Ⅱ表达。结果:番红染色ACLT+NS组软骨结构破坏明显,Mankin’s评分显著高于Sham组和ACLT+IB组(P <0.001);ACLT+IB组相对骨体积(BV/TV)、骨小梁数量(Tb.N)、骨小梁厚度(Tb.Th)等均明显高于ACLT+NS组(P<0.001),骨小梁分离度(Tb.Sp)则显著低于ACLT+NS (P <0.001);ACLT+NS组血清CTX-I、CTX-Ⅱ、IL-1、IL-6水平高于Sham组和ACLT+IB组(P <0.05);相比较于Sham组和ACLT+IB组,ACLT+NS组的MMP-13表达水平增高(P <0.01),LC3 Ⅱ表达降低(P <0.01)。结论:伊班膦酸钠能够下调MMP-13蛋白表达,并减少炎性因子IL-1和IL-6分泌,进而保护关节软骨,改善软骨下骨小梁微观结构,可能与自噬相关蛋白LC3 Ⅱ表达增高有关。Objective: To study the effects of ibandronate sodium on the cartilage and subchondral bone of anterior cruciate ligament transaction(ACLT) in rats with knee osteoarthritis and to explore the role of autophagy-related protein LC3 Ⅱ in the development of osteoarthritis. Methods: Thirty 3-month-old female SD rats were randomly divided into 3 groups(n = 10): sham operation group(SHAM), ACLT + NS group, and ACLT + IB group. Rats in ACLT + NS and ACLT + IB groups were undergone ACLT on the right knee. The rats of sham group were undergone sham operation. One week later, the rats of ACLT + IB group were given intraperitoneal injection of ibandronate sodium at a dosage of 10 μg/kg weekly for 12 weeks. In contrast, the ACLT + NS group received the same dose of NS. All rats were sacri?ced 13 weeks after operation. Safranin staining was used to observe the structure of articular cartilage. The degeneration of cartilage was graded by Mankin’s scale. The changes of subchondral bone were quantitatively analyzed by Micro-CT. The secretion of IL-1(Interleukin-1), IL-6(Interleukin-6), C-terminal cross-linking telopeptide of type I collagen(CTX-I) and C-terminal cross-linking telopeptide of type I collagen(CTX-Ⅱ) in serum was examined by ELISA. The protein expression level of matrix metalloproteinase-13(MMP-13) and LC3 Ⅱ was detected by Western blotting. Results: The destruction of articular cartilage was observed in ACLT + NS group. The mankin’s scores in ACLT + IB and Sham groups were significantly lower than that in ACLT + NS group(P < 0.001). Compared with ACLT + NS group, the trabecular bone relative volume(BV/TV), trabecular bone thickness(Tb.Th), trabecular bone number(Tb.N) of rats in ACLT + IB group increased signi?cantly(P < 0.001), while the trabecular bone separation(Tb.Sp) decreased signi?cantly(P < 0.001). Serum levels of CTX-I, CTX-Ⅱ, IL-1, IL-6 in ACLT + IB and Sham groups were signi?cantly lower than those in ACLT + NS group(P < 0.05). The expression of MMP-13 in ACLT + IB group was signi?cantly lowe

关 键 词：伊班膦酸钠 骨关节炎 软骨 软骨下骨 自噬相关蛋白LC3Ⅱ 

分 类 号：R684.3[医药卫生—骨科学]