Treatment of pulmonary fibrosis with siRNA against a collagen-specific chaperone HSP47 in vitamin A-coupled liposomes
Treatment of pulmonary fibrosis with siRNA against a collagen-specific chaperone HSP47 in vitamin A-coupled liposomes
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DOI：
10.1080/01902148.2017.1354946
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发表时间：
2017-01-01
影响因子：
1.7
通讯作者：
Takahashi, Hiroki
中科院分区：
文献类型：
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作者：
Otsuka, Mitsuo;Shiratori, Masanori;Takahashi, Hiroki
背景资料：肺纤维化是进行性间质性肺疾病（如特发性肺纤维化）的危及生命的病理状态。肌成纤维细胞在肺纤维化的发病机制中起重要作用。本研究旨在评估小干扰RNA（siRNA）对胶原特异性伴侣热休克蛋白47（HSP 47）的抑制作用。在博莱霉素（BLM）诱导的肺纤维化大鼠模型中，使用封装在维生素A偶联脂质体（VA-lip-siRNA HSP 47）中的针对HSP 47的siRNA，将siRNA优先递送至肌成纤维细胞。方法和结果：在第1天至第21天的预防性施用时间表和第15天至第35天的治疗性施用时间表下，用BLM或磷酸盐缓冲盐水的气管内注射处理雄性Sprague-Dawley大鼠，随后每周三次静脉内注射VA-lip-siRNA HSP 47。用免疫印迹法检测热休克蛋白47的表达。通过免疫荧光染色检测与6 '-羧基荧光缀合的VA-lip-siRNA HSP 47到肌成纤维细胞中的特异性递送。通过形态学和生物化学方法分析VA-lip-siRNA HSP 47对纤维化的影响。通过免疫荧光染色验证了VA-lip-siRNA HSP 47向BLM处理的大鼠中纤维化区域中的肌成纤维细胞的优先递送。VA-lip-siRNA HSP 47的处理明显抑制了BLM处理大鼠肺中HSP 47的表达并诱导了肌成纤维细胞的凋亡。BLM治疗大鼠的肺中羟脯氨酸水平和炎性细胞因子，以及支气管肺泡灌洗液中炎性细胞的数量显着抑制治疗。形态学评估显示，VA-lip-siRNA HSP 47在预防和治疗方案中也显著改善BLM处理的大鼠的形态学肺纤维化。结论：这些结果表明，VA-lip-siRNA HSP 47不仅在预防方面而且在治疗方面改善肺纤维化，因此，这种药物递送系统应该为难治性肺纤维化提供新的治疗。
Background: Pulmonary fibrosis is a life-threatening pathological state of progressive interstitial lung diseases, such as idiopathic pulmonary fibrosis. Myofibroblasts are known to play a critical role in the pathogenesis of pulmonary fibrosis. This study aimed to evaluate the inhibitory effect of a small interfering RNA (siRNA) on a collagen-specific chaperone heat shock protein 47 (HSP47). The siRNA was preferentially delivered to myofibroblasts in a bleomycin (BLM)-induced pulmonary fibrosis rat model using siRNA against HSP47, encapsulated in a vitamin A-coupled liposome (VA-lip-siRNA HSP47). Methods and Results: Male Sprague-Dawley rats were treated with an intratracheal injection of BLM or phosphate buffered saline followed by an intravenous injection of VA-lip-siRNA HSP47 three times per week under preventive administration schedules from day 1 to day 21 and therapeutic administration schedules from day 15 to day 35. The expression of HSP47 after the treatment was assessed by immunoblotting. The specific delivery of VA-lip-siRNA HSP47 conjugated with 6'-carboxyfluoresce into myofibroblasts was examined by immunofluorescence staining. The effect of VA-lip-siRNA HSP47 on fibrosis was analyzed by morphological and biochemical methods. Preferential delivery of VA-lip-siRNA HSP47 to myofibroblasts in fibrotic areas in BLM-treated rats was verified by immunofluorescence staining. Treatment of VA-lip-siRNA HSP47 clearly suppressed HSP47 expression and induced apoptosis of myofibroblasts in the lung of BLM-treated rats. Hydroxyproline levels and inflammatory cytokines in the lungs, and the number of inflammatory cells in the bronchial alveolar lavage of BLM-treated rats were significantly suppressed by the treatment. Morphological assessment showed that VA-lip-siRNA HSP47 also significantly improved the morphological pulmonary fibrosis of BLM-treated rats in both preventive and therapeutic schedules. Conclusions: These results suggest that VA-lip-siRNA HSP47 improves pulmonary fibrosis in not only preventive, but also therapeutic schedules, and thus, this drug delivery system should provide a novel therapy for refractory pulmonary fibrosis.