Tongluo Huatan capsule(通络化痰胶囊) improves cognitive function by regulating the endocytosis of N-methyl-D-aspartic acid receptors mediated by clathrin in a rat model of vascular dementia

OBJECTIVE:To explore the neuroprotective mechanisms of Tongluo Huatan capsule(THC)in a rat model of vascular dementia(VD).METHODS:A rat model of VD was established by repeated clamping of bilateral common carotid arteries with the intraperitoneal injection of sodium nitroprusside solution.VD rats were administered THC,memantine hydrochloride,or distilled water daily for 14 d after operation.Learning and memory abilities were assessed using the step-down passive avoidance test,novel object recognition(NOR)test,and Morris water maze(MWM)test.Pathological changes in the hippocampus were observed through hematoxylin and eosin and Nissl staining.The expression levels of clathrin,RAB5 B,andN-methyl-D-aspartic acid receptor 1(NMDAR1)were measured by immunohistochemistry staining,real-time quantitative polymerase chain reaction and Western blot.RESULTS:Rats in VD group showed impaired learning and memory abilities(step-down passive avoidance,NOR,and MWM)and abnormalities in neuronal morphology(light microscopy)in the hippocampus.The m RNA or protein expression levels of clathrin and RAB5 B were decreased,and NMDAR1 was increased in hippocampal tissues(P<0.05).Administration of THC promoted the learning and memory abilities and the morphological structure of hippocampal neurons in VD rats.Besides,THC enhanced m RNA or protein expression levels of clathrin and RAB5 B,and decreased NMDAR1(P<0.05).CONCLUSION:THC may improve cognitive functions by regulating the endocytosis of NMDA receptors mediated by clathrin.

Xinglou Chengqi Decoction improves neurological function in experimental stroke mice as evidenced by gut microbiota analysis and network pharmacology

The current study was designed to explore the brain protection mechanism of Xinglou Chengqi Decoction(XCD)based on gut microbiota analysis and network pharmacology. A transient middle cerebral artery occlusion(MCAO) model of mice was established, followed by behavioral evaluation, TTC and TUNEL staining. Additionally, to investigate the effects of gut microbiota on neurological function after stroke, C57BL/6 mice were treated with anti-biotic cocktails 14 days prior to ischemic stroke(IS) to deplete the gut microbiota. High-throughput 16S rDNA gene sequencing, metabonomics technique, and flow multifactor technology were used to analyze bacterial communities, SCFAs and inflammatory cytokines respectively. Finally, as a supplement, network pharmacology and molecular docking were applied to fully explore the multicomponent-multitarget-multichannel mechanism of XCD in treating IS, implicated in ADME screening, target identification, network analysis, functional annotation, and pathway enrichment analysis.We found that XCD effectively improved neurological function, relieved cerebral infarction and decreased the neuronal apoptosis.Moreover, XCD promoted the release of anti-inflammatory factor like IL-10, while down-regulating pro-inflammatory factors such as TNF-α, IL-17A, and IL-22. Furthermore, XCD significantly increased the levels of short chain fatty acids(SCFAs), especially butyric acid. The mechanism might be related to the regulation of SCFAs-producing bacteria like Verrucomicrobia and Akkermansia, and bacteria that regulate inflammation like Paraprevotella, Roseburia, Streptophyta and Enterococcu. Finally, in the network pharmacological analysis, 51 active compounds in XCD and 44 intersection targets of IS and XCD were selected. As a validation, components in XCD docked well with key targets. It was obviously that biological processes were mainly involved in the regulation of apoptotic process, inflammatory response, response to fatty acid, and regulation of establishment of endothelial barrier in GO enrichment. XCD can improve neurological function in experimental stroke mice, partly due to the regulation of gut microbiota. Besises, XCD has the characteristic of "multi-component, multi-target and multi-channel" in the treatment of IS revealed by network pharmacology and molecular docking.