Department of Nursing, Chung Hwa University of Medical Technology, Tainan Hsien 717, Taiwan.
Hum Gene Ther. 2009 Feb;20(2):147-58. doi: 10.1089/hum.2008.096.
In osteoarthritis (OA), inflammation and apoptosis are two important factors contributing to disease progression. As kallistatin can suppress inflammatory responses and reduce cell apoptosis, we investigated the therapeutic effect of kallistatin gene transfer in the rat model of OA by anterior cruciate ligament transection (ACLT). OA was induced in Wistar rats by ACLT in the knee of one hind limb. Adenoviral vector encoding human kallistatin (AdHKBP) was injected intraarticularly into the knee joints after ACLT. The viral effect on tissue was evaluated. The inflammatory responses and transgene expression were determined by immunoblot analysis, enzyme-linked immunosorbent assay, and immunohistochemistry. Apoptosis of chondrocytes was quantified by TUNEL assay. The effects of kallistatin in combination with hyaluronic acid (HA) on the medial femoral condyles and synovia were also assessed histologically. Inflammation trigged by the vectors was limited. Expression of human kallistatin after intraarticular injection was identified. Kallistatin gene transfer reduced the levels of interleukin-1beta and tumor necrosis factor-alpha in joints. Examination of gross morphology revealed that rats treated with AdHKBP had reduced severity of OA compared with control rats treated with adenoviral vector encoding green fluorescent protein (AdGFP). The protective effect of kallistatin on cartilage was accompanied by a decrease in apoptotic cells. Intraarticular administration of AdHKBP, when in conjunction with HA, significantly improved knee joint histologic scores. These results suggest that local administration of adenoviral vectors encoding kallistatin significantly suppressed OA progression, accompanied by reduction of inflammatory response and apoptosis. Thus, kallistatin gene therapy may be a potential treatment for OA.
在骨关节炎（OA）中，炎症和细胞凋亡是导致疾病进展的两个重要因素。由于卡利斯塔丁可以抑制炎症反应和减少细胞凋亡，我们通过前交叉韧带切断术（ACLT）研究了卡利斯塔丁基因转移在 OA 大鼠模型中的治疗效果。在一侧后肢的膝关节中通过 ACLT 诱导 Wistar 大鼠发生 OA。在 ACLT 后，将编码人卡利斯塔丁（AdHKBP）的腺病毒载体关节内注射到膝关节中。评估病毒对组织的影响。通过免疫印迹分析、酶联免疫吸附试验和免疫组织化学测定来确定炎症反应和转基因表达。通过 TUNEL 测定法量化软骨细胞的凋亡。还通过组织学评估了卡利斯塔丁与透明质酸（HA）联合对股骨内侧髁和滑膜的影响。载体引发的炎症受到限制。关节内注射后鉴定出人卡利斯塔丁的表达。卡利斯塔丁基因转移降低了关节中白细胞介素-1β和肿瘤坏死因子-α的水平。大体形态学检查显示，与用编码绿色荧光蛋白的腺病毒载体（AdGFP）治疗的对照大鼠相比，用 AdHKBP 治疗的大鼠 OA 严重程度降低。卡利斯塔丁对软骨的保护作用伴随着凋亡细胞的减少。与 HA 联合关节内给予 AdHKBP 显著改善了膝关节组织学评分。这些结果表明，局部给予编码卡利斯塔丁的腺病毒载体显著抑制了 OA 的进展，同时减少了炎症反应和凋亡。因此，卡利斯塔丁基因治疗可能是 OA 的一种潜在治疗方法。
