Anti-Inflammatory Effect of the Proteasome Inhibitor Bortezomib on Endotoxin-Induced Uveitis in Rats
Anti-Inflammatory Effect of the Proteasome Inhibitor Bortezomib on Endotoxin-Induced Uveitis in Rats
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DOI：
10.1167/iovs.12-9505
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发表时间：
2012-06-01
影响因子：
4.4
通讯作者：
Yang, Chang-Hao
中科院分区：
文献类型：
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作者：
Chen, Fang-Ting;Liu, Yi-Chun;Yang, Chang-Hao
目的。我们评估了蛋白酶体抑制剂博尔替佐米（Velcade）对内毒素诱导的大鼠葡萄膜炎（EIU）和脂多糖（LPS）刺激的RAW 264.7细胞的抗炎作用。采用足部注射LPS诱导Lewis大鼠EIU。各治疗组在LPS注射前30分钟给予mg -132 （10 mg/kg）或高剂量（0.2 mg/kg）或低剂量（0.05 mg/kg）硼替佐米。24小时后处死大鼠，观察组织炎症反应。采用PCR和Western blot检测fractalkine、MCP-1、ICAM-1和iNOS的表达水平。免疫组化（IHC）研究证实了促炎介质和核因子κ B (nf - κ B) p65在虹膜和睫状体中的表达。利用电泳迁移迁移试验（EMSA）评价NF-kappa B的dna结合活性。使用RAW 264.7细胞进行体外研究以验证结果。大剂量硼替佐米预处理可显著减轻EIU的炎症反应。在高剂量硼替佐米组和MG-132组中总是观察到炎症介质的表达降低，但在低剂量硼替佐米组中总是没有注意到。在高剂量硼替佐米或MG-132预处理的大鼠中，nf - κ B的dna结合活性降低。体外研究表明硼替佐米对lps刺激的RAW细胞具有剂量依赖性的抗炎作用，与体内实验结果一致。硼替佐米抑制EIU，可能是通过抑制NF-kappa B的激活，进而下调相关炎症基因的表达。蛋白酶体抑制可能是葡萄膜炎的一种潜在治疗策略。（Invest Ophthalmol Vis Sci. 2012;53:3682-3694） DOI: 10.1167/iovs.12-9505
PURPOSE. We evaluated the anti-inflammatory effect of bortezomib (Velcade), a proteasome inhibitor, on endotoxin-induced uveitis (EIU) in rats and lipopolysaccharide (LPS)-stimulated RAW 264.7 cells.METHODS. EIU was induced by footpad injection of LPS into Lewis rats. MG-132 (10 mg/kg), or high-dose (0.2 mg/kg) or low-dose (0.05 mg/kg) bortezomib was given 30 minutes before LPS injection in each treatment group. The rats were sacrificed 24 hours later to observe the inflammatory response in tissues. The expression levels of fractalkine, MCP-1, ICAM-1, and iNOS were evaluated by PCR and Western blot analysis. Immunohistochemical (IHC) studies were used to demonstrate the expression of pro-inflammatory mediators and nuclear factor-kappa B (NF-kappa B) p65 in the iris and ciliary body. The DNA-binding activity of NF-kappa B was evaluated using an electrophoretic mobility shift assay (EMSA). An in vitro study using RAW 264.7 cells was performed to verify the results.RESULTS. Pretreatment with high-dose bortezomib significantly attenuated the inflammatory response of EIU. Reduced expression of inflammatory mediators always was observed in the high-dose bortezomib and MG-132 groups, but invariably was not noted in the low-dose bortezomib group. Decreased DNA-binding activity of NF-kappa B was noted in those rats pretreated with high-dose bortezomib or MG-132. In vitro study demonstrated the dose-dependent anti-inflammatory effects of bortezomib in LPS-stimulated RAW cells, consistent with the results obtained in vivo.CONCLUSIONS. Bortezomib inhibits EIU, probably by inhibiting the activation of NF-kappa B, which in turn, down-regulates the expression of the associated inflammatory genes. Proteasome inhibition may be a potential treatment strategy for uveitis. (Invest Ophthalmol Vis Sci. 2012;53:3682-3694) DOI: 10.1167/iovs.12-9505