The present study is focused on the modulation of Mycobacterium bovis BCG-induced inflammatory response by poly-dispersed acid-functionalized single-walled carbon nanotubes (AF-SWCNTs) in macrophages. Flow cytometric and confocal microscopy studies indicated that both BCG and AF-SWCNTs were efficiently internalized by RAW 264.7 and MH-S macrophage cell lines and were essentially localized in the cytoplasmic area. BCG-induced production of reactive oxygen species (ROS) and nitric oxide by the two cell lines was significantly inhibited by AF-SWCNTs. Using RT-PCR technique, a marked decline was observed in the expression of BCG-induced pro-inflammatory genes COX-2, iNOS, TNF-α, IL-6, and IL-1β upon treatment with AF-SWCNTs. Results of gelatin zymography indicated that the AF-SWCNTs treatment also induced a marked decline in BCG-induced release of matrix metalloproteinases MMP-2 and MMP-9 by the two macrophage cell lines. The anti-inflammatory effect of AF-SWCNTs in downregulating BCG-induced inflammatory response was further validated in murine peritoneal macrophages. Treatment with AF-SWCNTs led to a steep decline in BCG-induced NO production in murine peritoneal macrophages in vitro as well as in vivo. Peritoneal macrophages isolated from mice treated with BCG and AF-SWCNTs had a significantly lower intracellular expression of COX-2 as compared to the peritoneal macrophages derived from mice treated with BCG alone. Taken together, our results demonstrate a potent anti-inflammatory effect of AF-SWCNTs in alleviating BCG-induced inflammatory responses in macrophages in vitro and in vivo.

本研究的重点是对牛分枝杆菌的调节BCG 通过多分散酸功能化单壁碳纳米管 (AF-SWCNT) 在巨噬细胞中诱导的炎症反应。流式细胞术和共聚焦显微镜研究表明，BCG 和 AF-SWCNT 都被 RAW 264.7 和 MH-S 巨噬细胞系有效内化，并且基本上定位于细胞质区域。AF-SWCNTs 显着抑制了 BCG 诱导的两种细胞系产生的活性氧 (ROS) 和一氧化氮。使用 RT-PCR 技术，在用 AF-SWCNTs 处理后，观察到卡介苗诱导的促炎基因 COX-2、iNOS、TNF-α、IL-6 和 IL-1β 的表达显着下降。明胶酶谱学结果表明，AF-SWCNTs 处理还诱导两种巨噬细胞系 BCG 诱导的基质金属蛋白酶 MMP-2 和 MMP-9 的释放显着下降。AF-SWCNTs 在下调 BCG 诱导的炎症反应中的抗炎作用在小鼠腹腔巨噬细胞中得到进一步验证。AF-SWCNTs 治疗导致小鼠腹腔巨噬细胞中 BCG 诱导的 NO 产生急剧下降体外和体内. 与来自单独用 BCG 治疗的小鼠的腹膜巨噬细胞相比，从用 BCG 和 AF-SWCNT 治疗的小鼠分离的腹膜巨噬细胞具有显着降低的 COX-2 细胞内表达。总之，我们的结果证明了 AF-SWCNT在体外和体内减轻 BCG 诱导的巨噬细胞炎症反应方面具有有效的抗炎作用。