Analysis of the regulating PI3K/AKT/mTOR signaling pathway and anti-apoptosis activity of Shenqi granule through Network Pharmacology and in vitro experiments

Abstract

Idiopathic membranous nephropathy (IMN), a common pathological type of nephrotic syndrome. Shenqi granule(SQ) is a traditional Chinese medical formula that has been used for decades to treat IMN, and there is a large amount of clinical data confirming its effectiveness,but the mechanism is unclear. This study explores the potential mechanisms and targets of action of SQ through network pharmacology and validates them through in vitro experiments and molecular docking techniques. Network pharmacology is a method that can determine how TCM works through pharmacokinetic evaluation, allowing us to study its molecular mechanisms. Through in vitro experiments, MPC5 cells are used to establish puromycin aminonucleoside (PAN)-induced podocytes damage models to extract cell protein, western blot detection signal pathway protein and related target proteins, molecular docking was performed for the validation. The network pharmacology study results indicate that SQ has 106 compounds, and 195 shared targets with MN. The treatment of IMN with SQ is mainly related to the apoptosis, PI3K/AKT/mTOR signaling pathway and other significant signaling pathways. In vitro experiments showed that SQ could effectively inhibit the activity of the PI3K/AKT/mTOR signaling pathway, increase the expression of Bcl2, and suppress the expression levels of apoptosis-related proteins such as Calaspase-3 and Bax in MPC5 cells. This study initially investigated the pharmacological effects of SQ, which effectively ameliorates IMN by potentially regulating the phosphorylation level of the AKT/mTOR pathway, inhibiting apoptotic activity, and restoring skeletal proteins.