Exploring the pharmacological mechanism of calculus bovis on cerebral ischemic stroke using a network pharmacology approach

Abstract Ethnopharmacological relevance Calculus bovis is commonly used in traditional Chinese medicine for the treatment of cerebrovascular diseases, with the role of clearing away heat and detoxification, detumulation and pain relief, such as the treatment of cerebral ischaemia, liver and gallbladder diseases and various inflammtion.However, the mechanism by which calculus bovis treats ischemic stroke is not well understood. Aim of the study In this study, the anti-inflammatory, anti-oxidative and antiapoptot-ic effects of calculus bovis on neurovascular units were studied, and the mechanism of in vitro cultured calculus bovis on neurovascular units was also discussed. Materials and methods Neurons, astrocytes, and endothelial cells were used to construct models of brain neurovascular units in vitro. The oxygen–glucose deprivation/reoxygenation and glucose (OGD/R) model was used to detect the efficacy of in vitro cultured calculus bovis, inflammatory factors, oxidative stress, and apoptosis. The expression of ZO-1, Occludin, Claudin-5, HIF-1α, VEGF, PI3K, Akt, Bax, Bcl-2, and Caspase-3 were detected. Results In vitro cultured calculus bovis protects the blood-brain barrier, repair tight junction proteins, increases the protein expression of ZO-1, Occludin and Claudin-5, maintains the TEER value, repairs damaged endothelial cells, increases the activity of γ-GT, reduce LDH, reduces inflammatory injury, and reduces the level of TNF-α, LI-6, and IL-1β. In vitro cultured calculus bovis reduces oxidative stress damage, reduces NO, and improves SOD activity. In vitro cultured calculus bovis protects neurons through anti-apoptotic activity, reduces the apoptotic proteins Bax and Caspase-3, increases the protein expression level of Bcl-2, and protects brain neurovascular units through the HIF/VEGF and PI3K/Akt signalling pathways. Conclusion In summary, the protective effect of calculus bovis on neurovascular units can be realized through anti-oxidative stress, anti-inflammatory and antiapoptotic.The mechanism of action of in vitro cultured calculus bovis on ischemic stroke involves multiple targets and signalling pathways. PI3K/Akt, HIF-1α and VEGF pathways have been proved to be effective in protecting neurovascular units in the brain.