Exploring the in vitro anti-inflammatory effects of total steroids extract purified from the calyx of Physalis alkekengi L. var franchetii through an integrated approach combining network pharmacology, molecular docking, and experimental verification

Physalis alkekengi L. var. franchetii (PAFM) has traditionally been used for treatment of acute and chronic bronchitis, pharyngitis, and dysuria with long history. This study aimed to develop an efficient purification process of total steroids from the calyx of PAFM and demonstrate the anti-inflammatory activity of the steriods-riched extract. The optimal purification process by AB-8 macroporous resin was obtained which increased the content of total steroids of the extract of PAFM (PAPS) from 5.47 % to 52.86 %. A total of 13 compounds was identified by UPLC-Q-Exactive-Orbitrap MS in PAPS, including 6 physalins, 3 flavonoids, 1 fatty acid and 3 sucrose fatty acid esters. A total of 24 core targets were predicted by network pharmacology analysis, mainly involving MAPK1, PTGS2, MMP9 and NF-κB. Molecular docking was performed to confirm the affinity of key proteins for main components. Anti-inflammatory study showed that PAPS could dose-dependently inhibit the secretion of NO and TNF-α, IL-1β and IL-6 in LPS-induced RAW 264.7 cells. It exerted anti-inflammatory effects by downregulating the expression of proteins including NF-κB, and COX-2, suppressing the phosphorylation of ERK. The results explore an effective approach to preparing total steriods extract of PAFM. The anti-inflammatory activity of PAPS demonstrated that it has the therapeutic potential in inflammatory diseases. • The total steriod content of the purified extract from PAFM raised from 5.47 % to 52.86 %. • 13 bioactive compounds including 6 physalins were identified by UPLC-Q-Exactive-Orbitrap MS in PAPS. • Network pharmacology identified 24 core anti-inflammatory targets (MAPK1, PTGS2, MMP9), docking-validated favorable binding. • PAPS dose-dependently inhibits inflammatory factors in LPS-induced RAW264.7 cells via downregulating NF-κB/COX-2 and suppressing ERK phosphorylation.