Doh Kyung-Oh, Jung Hyun-Kyung, Moon Ik-Jae, Kang Hyun-Gu, Park Jeong-Hoh, Park Jong-Gu
Department of Physiology, Dongguk University College of Medicine, Gyeongju, Korea.
Int J Mol Med. 2008 Jan;21(1):33-9.
Transforming growth factor-beta1 (TGF-beta1) is an important mediator of tissue fibrosis, including liver cirrhosis. Ribbon-type antisense oligonucleotide to TGF-beta1 (TGF-beta1 RiAS) was designed and combined with cationic peptide derived from the nuclear localization signal of human immunodeficiency virus-1 Tat protein for enhanced cellular uptake. When Hepa1c1c7 cells were transfected with TGF-beta1 RiAS, the level of TGF-beta1 mRNA was reduced by >70%. TGF-beta1 RiAS, mismatched RiAS, and normal saline were each injected into mice via the tail vein, beginning the week after intraperitoneal CCl4 injection and continuing for 7 weeks, in order to determine whether TGF-beta1 RiAS prevents the fibrotic changes induced by the CCl4 injection. After 8 weeks of the experiment, all of the mice treated with TGF-beta1 RiAS survived, compared to 50% of the control group and 65% of the mismatched RiAS-treated group. Upon examining the biochemical effects on the liver, TGF-beta1 mRNA levels were reduced significantly only in the TGF-beta1 RiAS-treated group. Immunohistochemical studies showed a reduced accumulation of collagen and alpha-smooth muscle actin. Our experimental results suggest that ribbon antisense to TGF-beta1, with efficient uptake, effectively blocks the expression of TGF-beta1 and prevents fibrosis of the liver.
转化生长因子-β1（TGF-β1）是包括肝硬化在内的组织纤维化的重要介质。设计了针对TGF-β1的带状反义寡核苷酸（TGF-β1 RiAS），并将其与源自人类免疫缺陷病毒1型Tat蛋白核定位信号的阳离子肽结合，以增强细胞摄取。当用TGF-β1 RiAS转染Hepa1c1c7细胞时，TGF-β1 mRNA水平降低了70%以上。为了确定TGF-β1 RiAS是否能预防由四氯化碳注射诱导的纤维化变化，在腹腔注射四氯化碳后的第一周开始，通过尾静脉分别向小鼠注射TGF-β1 RiAS、错配的RiAS和生理盐水，并持续7周。实验8周后，所有接受TGF-β1 RiAS治疗的小鼠存活，而对照组和错配RiAS治疗组的存活率分别为50%和65%。在检查对肝脏的生化影响时，仅在TGF-β1 RiAS治疗组中TGF-β1 mRNA水平显著降低。免疫组织化学研究显示胶原蛋白和α-平滑肌肌动蛋白的积累减少。我们的实验结果表明，具有有效摄取能力的TGF-β1带状反义寡核苷酸能有效阻断TGF-β1的表达并预防肝脏纤维化。
