Hyperglycemic mice were treated with TNTL. The anti-hyperglycemic effect of TNTL was validated to confirm the biological consistency of the herbal products from batches. Improvement of DR by TNTL was examined by various assays on the retina. Next-generation transcriptome sequencing and cytokine array was used to identify the therapeutic targets. In vitro study was performed to validate the target.
We observed that TNTL at its high doses possessed anti-hyperglycemic effect in murine type I diabetic model, while at its doses without reducing blood glucose, it suppressed DR incidence. TNTL restored the blood-retina barrier integrity, suppressed retinal neovascularization, and attenuated the retinal ganglion cell degeneration. Transcriptomic analysis on the retina tissue of hyperglycemic mice with or without TNTL revealed that the inflammatory retina microenvironment was significantly repressed. TNTL treatment suppressed pro-inflammatory macrophages in the retina, which resulted in the inactivation of endothelial cell migration, restoration of endothelial cell monolayer integrity, and prevention of leakage. Cytokine array analysis suggested that TNTL could significantly inhibit the secretion of MIP1γ from pro-inflammatory macrophages. Prevention of endothelial dysfunction by TNTL may be mediated by the inhibition of MIP1γ/CCR1 axis. More specifically, TNTL suppressed MIP1γ release from pro-inflammatory macrophages, which in turn inhibited the activation of CCR1-associated signaling pathways in endothelial cells.
Our findings demonstrated that TNTL might be an alternative treatment to DR, and the primary source of potential drug candidates against DR targeting MIP1γ/CCR1 axis in the retinal microenvironment.
用TNTL处理高血糖小鼠。验证了TNTL的抗高血糖作用,以确认来自批次的草药产品的生物一致性。通过视网膜上的各种测定检查了TNTL对DR的改善。使用下一代转录组测序和细胞因子阵列来鉴定治疗靶标。进行体外研究以验证靶标。
我们观察到高剂量的TNTL在小鼠I型糖尿病模型中具有抗高血糖作用,在不降低血糖的情况下,它抑制了DR的发病率。TNTL恢复了血-视网膜屏障的完整性,抑制视网膜新生血管,减轻视网膜神经节细胞变性。对有或没有TNTL的高血糖小鼠的视网膜组织的转录组学分析显示,炎性视网膜微环境被显着抑制。TNTL治疗抑制视网膜中的促炎巨噬细胞,导致内皮细胞迁移失活,内皮细胞单层完整性的恢复,和防止泄漏。细胞因子阵列分析表明,TNTL可以显着抑制促炎巨噬细胞分泌MIP1γ。TNTL预防内皮功能障碍可能是通过抑制MIP1γ/CCR1轴介导的。更具体地说,TNTL抑制促炎巨噬细胞释放MIP1γ,这反过来又抑制了内皮细胞中CCR1相关信号通路的激活。
我们的研究结果表明,TNTL可能是DR的替代疗法,以及在视网膜微环境中靶向MIP1γ/CCR1轴的DR的潜在候选药物的主要来源。