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目的研究重组腺病毒介导的人内皮抑素(nAD-GFP-ES)对大鼠Ⅱ型胶原诱导型关节炎(CIA)炎症及细胞因子的影响。方法首先,扩增并纯化重组腺病毒。然后,建立CIA模型,自造模次日治疗组给予rAD-GFP-ES(1×10~(11)pfu·kg~(-1)·周~(-1)×4周),各组大鼠每周进行一次关节炎指数(AI)评分,于造模第4周取血清行Western Blot检测大鼠体内内皮抑素(ES)、白细胞介素(IL)-1D、肿瘤坏死因子(TNF)-α的表达并计算相对含量。结果①纯化后的rAD-GFP-ES和rAD-GFP滴度分别为：6．6×10~(12) pfu/m1．4．8×10~(12)pfu/ml；吸光度(A)_(260nm)/A_(2180nm)均＞1．3。②感染rAD-GFP-ES的大鼠体内ES获得了稳定表达,是非模型组的2．4倍。③感染rAD-GFP-ES可以降低大鼠AI评分及IL-1β、TNF-α的表达。结论①rAD-GFP- ES注射可使ES基因在大鼠体内获得稳定表达。②感染rAD-GFP-ES能明显降低模型大鼠的AI评分。③IL-1β、TNF-α共同促进CIA大鼠炎症的形成和发展,rAD-GFP-ES可能通过抑制IL-1β、TNF-α的表达而发挥抗炎的作用。
Objective To investigate the effect of recombinant adenovirus mediated human endostatin (nAD-GFP-ES) on inflammation and cytokines in rat model of collagen induced arthritis (CIA). Methods Firstly, the recombinant adenovirus was amplified and purified. Then, the CIA model was established, and rAD-GFP-ES (1 × 10-11 pfu · kg -1 · week -1 × 4 weeks) The rats were sacrificed once a week for evaluation of the AI index. Serum levels of endostatin (ES), interleukin (IL) -1D and tumor necrosis factor (TNF) -α expression and calculate the relative content. Results ① The titer of purified rAD-GFP-ES and rAD-GFP were 6.6 × 10 ~ (12) pfu / ml.4.8 × 10 ~ (12) pfu / ml respectively; (260nm) / A 2180nm> 1.3. ② In vivo, ES of rats infected with rAD-GFP-ES was stably expressed, which was 2.4 times higher than that of non-model group. ③ Infection of rAD-GFP-ES can reduce the AI score of rats and the expression of IL-1β and TNF-α. Conclusion ① ESR gene can be stably expressed in rat after injection of rAD-GFP-ES. ② Infection with rAD-GFP-ES significantly reduced the AI score in model rats. IL-1β and TNF-α can promote the formation and development of inflammation in CIA rats. RAD-GFP-ES may play an anti-inflammatory role by inhibiting the expression of IL-1β and TNF-α.