Targeted and stimuli-responsive gene/drug synergistic therapeutic system based on pcDNA3.1-Mmp13 and hydroxychloroquine for the effective treatment of liver fibrosis in mice

Liver fibrosis is a pathological process of abnormal proliferation of extracellular matrix (ECM) in liver. The activation of hepatic stellate cells (HSCs) is the main driving factor to form ECM in liver fibrosis. Deposition of ECM could further reactivate HSCs to result in a vicious cycle to promote the deterioration of liver fibrosis. It is of great significance to explore effective synergistic strategies not only inhibiting HSCs activation but also degrading the already formed ECM for the treatment of liver fibrosis. The targeted and stimuli-responsive gene/drug synergistic therapeutic system HCQ/p Mmp13 @PR based on the plasmid of pcDNA3.1- Mmp13 (p Mmp13 ) and hydroxychloroquine (HCQ) was constructed by using gene vector polyethyleneimine-polyethylene glycol-retinol (PR) modified with retinol (ROL) for targeting aHSCs. HCQ/p Mmp13 @PR had good stability, gene protection ability, targeting ability to aHSCs and biosafety. HCQ/p Mmp13 @PR had the characteristic of stimuli-responsive release of HCQ and p Mmp13 by intracellular high glutathione (GSH) environment, thus producing effective synergistic anti-liver fibrosis performance. The immunofluorescence, immunohistochemistry, western blot and flow cytometry experiments confirmed the synergistic therapeutic effect in vitro and in vivo. HCQ/p Mmp13 @PR could not only inhibit the autophagy to reduce the activation of HSCs, but also promote the expression of matrix metalloproteinase-13 (MMP-13) to degrade ECM to improve liver fibrosis. HCQ/p Mmp13 @PR co-delivery system has a good synergistic effect on the treatment of liver fibrosis, which provides the clinical experimental basis for the development of anti-liver fibrosis drugs.