Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbits.
Effects of GGCX overexpression on anterior cruciate ligament transection-induced osteoarthritis in rabbits.
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DOI：
10.3892/mmr.2017.8304
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发表时间：
2017-11
影响因子：
3.4
通讯作者：
Xiaolin Fu;Ruiyun Qiu;Chunfang Tang;Xiaomei Wang;Xigao Cheng;Ming Yin
中科院分区：
文献类型：
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作者：
Xiaolin Fu;Ruiyun Qiu;Chunfang Tang;Xiaomei Wang;Xigao Cheng;Ming Yin
骨关节炎（OA）缺乏有效的治疗方法。γ-谷氨酰羧化酶（GGCX）是调节软骨基质Gla蛋白（MGP）羧化的关键酶。GGCX过表达是否能预防OA仍不清楚。本研究旨在探讨GGCX过表达对日本白色兔前交叉韧带横断（ACLT）诱导的OA的影响及其机制。采用ACLT手术建立兔OA模型。将48只家兔随机分为4组：假手术组、骨关节炎模型+ GGCX过表达质粒组、骨关节炎模型+生理盐水组和骨关节炎模型+空载体组。采用ELISA、免疫组化、逆转录-定量聚合酶链反应（RT-PCR）和蛋白质印迹法（Western blotting）分别检测非羧化MGP（ucMGP）、羧化MGP（cMGP）、基质金属蛋白酶（MMP）-13、X型胶原、II型胶原、肿瘤坏死因子（TNF）-α和白细胞介素（IL）-1 β的表达。分别通过Giemsa和番红O-fast绿色染色评估胫骨软骨的形态学变化。与假手术对照组相比，OA模型组GGCX表达明显降低。通过注射编码GGCX的慢病毒携带的过表达质粒增加GGCX表达。GGCX过表达改善了关节软骨中的ATL C诱导的损伤。OA模型组cMGP和II型胶原表达明显降低，ucMGP、X型胶原、MMP-13、IL-1β和TNF-α表达明显升高。值得注意的是，这些表达水平在OA模型兔中被GGCX过表达逆转。本研究的结果表明，在OA模型兔中GGCX的表达降低，而GGCX的过表达能够促进MGP的羧化，减轻炎症，降低MMP-13的表达并调节胶原的表达。结果还表明，GGCX可以作为OA的治疗靶点。
Effective therapeutic methods for osteoarthritis (OA) are lacking. γ‑glutamyl carboxylase (GGCX) is a key enzyme that regulates carboxylation of cartilage matrix Gla protein (MGP). Whether GGCX overexpression protects against OA remains unknown. The aim of the present study was to explore the effects of GGCX overexpression on anterior cruciate ligament transection (ACLT)‑induced OA and its mechanisms in Japanese white rabbits. ACLT surgery was used to establish an OA model in rabbits. A total of 48 rabbits were randomly divided into 4 groups: Sham, OA model + GGCX overexpression plasmid, OA model + saline and OA model + empty vector. The expression of uncarboxylated MGP (ucMGP), carboxylated MGP (cMGP), matrix metalloproteinase (MMP)‑13, collagen type X, collagen type II, tumor necrosis factor (TNF)‑α and interleukin (IL)‑1β were detected by ELISA, immunohistochemistry, reverse transcription‑quantitative polymerase chain reaction and western blotting. Morphological changes to tibial cartilage were assessed by Giemsa and safranin O‑fast green staining, respectively. Compared with the Sham control, GGCX expression was significantly decreased in the OA Model group. GGCX expression was increased by injection of a lentivirus‑carried overexpression plasmid that encoded GGCX. GGCX overexpression ameliorated ATLC‑induced damage in articular cartilage. OA Model rabbits exhibited significantly decreased expression levels of cMGP and collagen type II, and increased expression of ucMGP, collagen type X, MMP‑13, IL‑1β and TNF‑α. Notably, these expression levels were reversed by GGCX overexpression in OA Model rabbits. Results from the present study indicated that GGCX expression was decreased in OA Model rabbits, whereas overexpression of GGCX was able to promote carboxylation of MGP, reduce inflammation, decrease MMP‑13 expression and regulate collagen expression. The results also indicated that GGCX may serve as a therapeutic target for OA.