Penicillamine Combined with Gandou Decoction Regulates Caspase11-GSDMD Pathway to Inhibit Pyroptosis in Wilson Disease Model
Objective To investigate the inhibitory effects and potential mechanisms of penicillamine combined with Gandou Decoction on neuronal pyroptosis in a Wilson disease(WD)model.Methods The in vitro experiments included the following groups,i.e.,the normal group,model group(CuCl2 10 µ mol/L),control group(WEL 20 μ mol/L),and WEL group(CuCl2 10 µ mol/L+WEL 20 µ mol/L).And the in vivo experiments,included the following groups,i.e.,the normal group(0.9％NaCl solution 0.2 mL·10 g-1·d-1),model group(0.9％NaCl solution 0.2 mL·10 g-1·d-1),Gandou Decoction group(0.2 mL·10 g-1·d-1),penicillamine group(0.1 g·kg-1·d-1),and combination treatment group(penicillamine 0.1 g·kg-1·d-1+Gandou Decoction 0.2 mL·10 g-1·d-1).After 4 weeks of administration,motor function was assessed through behavioral experiments,and striatal tissues were collected.Neuronal damage was observed using Fluoro-Jade B(FJB)staining,ultrastructural changes were examined by transmission electron microscopy(TEM),copper content was measured using inductively coupled plasma mass spectrometry(ICP-MS),and microglial activation was assessed by immunofluorescence staining.The expression levels of pyroptosis-related genes and proteins in cells and striatal tissues were determined by RT-qPCR and Western Blot.Results In the in vivo experiments,compared to the normal group,the model group exhibited reduced motor function,increased FJB-positive cells in the striatum(P＜0.01),neuronal swelling,membrane pore formation,microglial activation(P＜0.01),and elevated expression levels of pyroptosis-related genes and proteins(P＜0.05).Compared to the model and penicillamine groups,the combination treatment group showed improved motor function,reduced FJB-positive cells(P＜0.01),intact neuronal cell membranes,intact neuronal cell membranes,and decreased microglial activation(P＜0.01),and lowered expression levels of pyroptosis-related genes and proteins(P＜0.05).In the in vitro experiments,the model group exhibited elevated expression levels of pyroptosis-related genes and proteins compared to the normal group(P＜0.05).The WEL group showed reduced expression levels of pyroptosis-related genes and proteins compared to the model group(P＜0.05).Conclusion Penicillamine combined with Gandou Decoction can effectively reduce neuronal damage in the WD model,and its mechanism is associated with the inhibition of Caspase11-GSDMD pathway-mediated neuronal pyroptosis.
Wilson's diseaseneuronal pyroptosisCaspase11-GSDMD pathwayGandou Decoctionpenicillamineintegrative medicine
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