A network pharmacology approach and experimental validation to investigate the protection mechanism of Ganoderma lucidum (Leyss. ex Fr.) Karst spore for acute kidney injury through induction of apoptosis via p53/Caspase 3 signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE

According to the Sheng Nong's herbal classic, one of the components Ganoderma lucidum (Leyss. ex Fr.) "benefits waterways and kidney qi". The reproductive cells of Ganoderma lucidum (Leyss. ex Fr.) Karst spore (GLS) has richer active substances, and the protective mechanism against acute kidney injury (AKI) needs to be clarified.

PURPOSE

This study dedicates to evaluate the effect of GLS on AKI, explores activation of the p53 signaling pathway, and investigates potential apoptosis-regulating mechanism.

METHODS

Gentamicin (GM) was used to establish an AKI rat model by intraperitoneal injection followed by the GLS intervention. The therapeutic efficacy of the GLS was evaluated based on function indices in rats. Network pharmacology was used to identify key proteins for the GLS intervention in AKI rats and bioinformatics analysis were performed. The initiation of the p53 signaling pathway was verified by molecular biology and surface enhanced raman spectroscopy (SERS).

RESULTS

UPLC-Q-Tof MS was carried to characterize the triterpene saponin analogs extracted from the GLS. The molecular docking results of the protein Caspase 3 showed that Autodock Binding energy was 8.86, and PyRx Binding Affinity was 9.6. Successfully established an AKI rat model. Compared with the GM group, the serum renal function markers in GLS dose groups were decreased; the degree of renal tissue lesions, renal tubular injury and inflammatory cell infiltration in GLS dose groups were significantly reduced, the expression of p53, Cytochrome C, BAX, Caspase-3 and Cleaved Caspase-3 proteins in the renal tissues of GLS dose groups were down-regulated, and the expression of BcL-2 protein was up-regulated. TUNEL staining and SERS also showed that GLS significantly inhibited the hyperapoptosis of kidney tissue caused by GM. This is the first study applying the GLS to treat GM-induced AKI.

CONCLUSION

This study was innovatively proposed that the GLS exerts a protective effect against AKI by inhibiting p53 overexpression. In addition, SERS was firstly applied to the spectroscopic observation of renal apoptotic tissues, and the results indicated that the GLS had a role in down-regulating excessive apoptosis in the injured kidney.