摘要：

Background: Diabetic retinopathy (DR) is a vision-threatening microvascular complication of diabetes mellitus. Chronic inflammation and endothelial dysfunction are critical factors in the disease's pathogenesis. Consequently, interventions developed to reduce retinal inflammation are anticipated to be beneficial for both the prevention and treatment of DR. In the present study, we developed a unique class of drugless peptide-based nanohybrids with potent anti-inflammatory activities and investigated their therapeutic efficacy for treating DR in an oxygen-induced retinopathy (OIR) mouse model and a streptozotocin (STZ)-induced diabetic mouse model. Methods: Hexapeptides were applied to modify gold nanoparticles to form the drugless peptide-based nanohybrids (P12). We then examined the physicochemical properties and anti-inflammatory activities of P12 in HUVECs and BV2 cells and identified the critical amino acids for this novel bioactivity. The intravitreal and retro-orbital injections were applied to determine the optimal retinal delivery route for P12. The therapeutic efficacy of P12 in treating DR were investigated using both the OIR model and STZ-induced diabetic model. Through immunohistochemistry and flow cytometry analyses, we identified the major cells that internalize P12 in the retina. Furthermore, in vitro experiments were used to explore the underlying molecular mechanisms for the anti-inflammatory activities of P12. Results: We found that P12 exhibited potent anti-inflammatory effects in both HUVECs and BV2 cells. In addition, P12 can be efficiently delivered to the retina via intravitreal injection. Intravitreally injected P12 significantly improved early DR symptoms including vascular leakage and pericyte loss in STZ-induced diabetic mice. It also suppressed pathological neovascularization and retinal hemorrhage in OIR mice. Importantly, we found that intravitreally injected P12 was mainly taken up by microglial and endothelial cells, leading to reduced retinal endothelium inflammation and microglial activation in DR animal models. Mechanistic studies revealed that P12 potently inhibited several TLR4 downstream signaling pathways, such as NF-κB, JNK, and P38 MAPK, in both endothelial and microglial cells. This effect is due to the capacity of P12 in blocking the endosomal acidification process that governs the endosomal TLR signaling transduction. Conclusions: Our findings suggest that local injection of properly designed, drugless, peptide-based nanohybrids can serve as a safe and effective anti-inflammatory nanomedicine for treating DR.

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