Multi -dimensional Mechanism Analysis of Choerospondias axillaris (Roxb.) Burtt et Hill in Treating Kidney Stones: Network Pharmacology, Molecular Docking and In Vitro Experimental Verification

Meiqi Qiu ^* Senhua Li Yu Zhang Jiaoxia Yan Shiting Qin Xijing Yin Yujun Li Chunhui Zeng ^* Ke Yang ^*

• Guangxi University of Chinese Medicine, Nanning, China

Results: Through network pharmacology analysis, 9 active ingredients were obtained, namely (-)-taxifolin, naringenin, (-)-catechin, quercetin, bis[(2S)-2-ethylhexyl] benzene-1,2-dicarboxylate, (2R)-5,7-dihydroxy-2-(4-hydroxyphenyl) chroman-4one, beta-sitosterol, ellagic acid, kaempferol. There were 272 protein targets for the active ingredients and 3525 for diseases. After intersecting the two sets, 187 potential targets were identified. PPI network analysis revealed that the top five core targets were AKT1, IL6, TNF, TP53, and IL-1β. GO analysis indicated that the treatment of kidney stones with CA was involved in biological processes like the response to oxidative stress and regulation of inflammatory response. KEGG prediction suggested that the treatment of kidney stones with CA was closely associated with signaling pathways such as NF-κB and MAPK. Molecular docking results demonstrated that five key active ingredients (quercetin, kaempferol, (-)-catechin, β-sitosterol and naringenin) exhibited good binding ability with their corresponding core targets. The results of two-dimensional CaOx agar gel system showed that the CA-L group significantly decreased the aggregation of COM crystals. In the CA-M and CA-H groups, the crystals mainly existed in the form of Calcium Oxalate Dihydrate (COD), which was readily excreted with urine, causing minimal damage to renal epithelial cells. Moreover, the crystal surface area was significantly smaller compared that of the model group. CA could protect cells damaged by COM crystals by increasing SOD activity, reducing ROS levels, and decreasing lactate dehydrogenase (LDH) leakage. Simultaneously, CA downregulated the expression of inflammatory proteins such as NLRP3, Caspase-1, IL-1β, as well as the expression of OPN protein, which promotes crystal adhesion. Conclusion: CA can attenuate the damage and adhesion of COM crystals to cells through multiple mechanisms. These include enhancing the cellular antioxidant capacity, regulating the activation of the NLRP3 inflammasome , reducing the expression of the crystal adhesion protein OPN, and preventing the further aggregation or mineralization of CaOx crystals. Thus, CA achieves the objective of treating kidney stones.