CD44 Receptor-Targeted and Reactive Oxygen Species-Responsive H2S Donor Micelles Based on Hyaluronic Acid for the Therapy of Renal Ischemia/Reperfusion Injury

Abstract

For the therapy attenuating renal ischemia-reperfusion (IR) injury, a novel drug delivery system was urgently needed, which could precisely deliver drugs to the pathological renal tissue. Here, we have prepared new nanomaterials with a reactive oxygen species (ROS)-responsive hydrogen sulfide (H₂S) donor and hyaluronic acid that targets CD44 receptor. The novel material was synthesized and characterized via related experiments. Then, rapamycin was loaded, which inhibited kidney damage. In the in vitro study, we found that the micelles had ROS-responsiveness, biocompatibility, and cell penetration. In addition, the experimental results showed that the intracellular H₂S concentration after administration was threefold higher than that of the control group. The western blot assay revealed that they have anti-inflammatory effects via H₂S donor blocking the NF-κB signaling pathway. Consequently, the rising CD44 receptor-targeting and ROS-sensitive H₂S donor micelles would provide a promising way for renal IR injury. This work provides a strategy for improving ischemia/reperfusion injury for pharmaceuticals.

Figure 1

RAP release mechanism diagram of ROS-sensitive, H₂S-responsive, and CD44 receptor-targeting nanoparticles.

Figure 2

Chemical synthesis of HATM.

Figure 3

Characterization of HATM. (A) ¹H-NMR spectrum of HATM. (B) Particle size, zeta potential, and TEM picture of HATM. Scale bars, 100 nm. (C) ROS-sensitive drug releases of HATM in vitro.

Figure 4

(A,B) Cytotoxicity of HATM@RAP for RAW264.7 cells and renal mesangial cells at 12 and 24 h. (C–F) Cell uptake of HATM@RAP for RAW264.7 cells and renal mesangial cells. (C) Concentration-dependent study of HATM in RAW264.7 cells. (D) Time-dependent study in RAW264.7 cells. (E) Result of the concentration-dependence study for renal mesangial cells. (F) Time-dependent study to renal mesangial cells. (G) Concentration of H₂O₂ in RAW264.7 cells after various groups (n = 3, mean ± SD). (H) Concentration of H₂S in RAW264.7 cells after various groups (n = 3, mean ± SD). (I) Representative Western blot bands of NF-κB and p-NF-κB after various groups. (J) Quantitative data of western blot of NF-κB and p-NF-κB (n = 3, mean ± SD). *P < 0.05, **P < 0.01.

Figure 5

H&E stained sections of liver, heart, spleen, and lungs after various groups.

Figure 6

HATM@RAP could improve renal function in renal IRI rats. (A–C) Concentration of urea nitrogen (A), creatinine (B), and urinary protein (C) after various groups.*P < 0.05, **P < 0.01. (D) H&E, PAS, and TUNEL stains of each group in kidneys.