[Experimental study of cardioprotective effects of Cinnamomi Ramulus and Cinnamomi Cortex formula granules on myocardial ischemia/reperfusion injury in rats based on efficacy of "warming and coordinating heart Yang"].

This study aimed to parallelly investigate the cardioprotective activity of Cinnamomi Ramulus formula granules(CRFG) and Cinnamomi Cortex formula granules(CCFG) against acute myocardial ischemia/reperfusion injury(MI/RI) and the underlying mechanism based on the efficacy of "warming and coordinating the heart Yang". Ninety male SD rats were randomly divided into a sham group, a model group, CRFG low and high-dose(0.5 and 1.0 g·kg~(-1)) groups, and CCFG low and high-dose(0.5 and 1.0 g·kg~(-1)) groups, with 15 rats in each group. The sham group and the model group were given equal volumes of normal saline by gavage. Before modeling, the drug was given by gavage once a day for 7 consecutive days. One hour after the last administration, the MI/RI rat model was established by ligating the left anterior descending artery(LAD) for 30 min ischemia followed by 2 h reperfusion except the sham group. The sham group underwent the same procedures without LAD ligation. Heart function, cardiac infarct size, cardiac patho-logy, cardiomyocyte apoptosis, cardiac injury enzymes, and inflammatory cytokines were determined to assess the protective effects of CRFG and CCFG against MI/RI. The gene expression levels of nucleotide-binding oligomerization domain-like receptor family pyrin domain protein 3(NLRP3) inflammasome, apoptosis-associated speck-like protein containing a CARD(ASC), cysteinyl aspartate specific proteinase-1(caspase-1), Gasdermin-D(GSDMD), interleukin-1ß(IL-1ß), and interleukin-18(IL-18) were determined by real-time quantitative polymerase chain reaction(RT-PCR). The protein expression levels of NLRP3, caspase-1, GSDMD, and N-GSDMD were determined by Western blot. The results showed that both CRFG and CCFG pretreatments significantly improved cardiac function, decreased the cardiac infarct size, inhibited cardiomyocyte apoptosis, and reduced the content of lactic dehydrogenase(LDH), creatine kinase MB isoenzyme(CK-MB), aspartate transaminase(AST), and cardiac troponin Ⅰ(cTnⅠ). In addition, CRFG and CCFG pretreatments significantly decreased the levels of IL-1ß, IL-6, and tumor necrosis factor-α(TNF-α) in serum. RT-PCR results showed that CRFG and CCFG pretreatment down-regulated the mRNA expression levels of NLRP3, caspase-1, ASC, and downstream pyroptosis-related effector substances including GSDMD, IL-18, and IL-1ß in cardiac tissues. Western blot revealed that CRFG and CCFG pretreatments significantly decreased the protein expression levels of NLRP3, caspase-1, GSDMD, and N-GSDMD in cardiac tissues. In conclusion, CRFG and CCFG pretreatments have obvious cardioprotective effects on MI/RI in rats, and the under-lying mechanism may be related to the inhibition of NLRP3/caspase-1/GSDMD signaling pathway to reduce the cardiac inflammatory response.

[Mechanism of Jingfang Granules in relieving alcohol and protecting liver based on bioinformatics technology].

The present study explored the potential mechanism of Jingfang Granules in relieving alcohol and protecting liver by network pharmacology and molecular docking and verified the effects and related pathways by animal experiments. The active components of Jingfang Granules were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Targets of drugs and diseases were obtained from PubChem, Swiss Target Prediction and CTD. The common targets were uploaded to STRING to plot the protein-protein interaction(PPI) network. The core targets were screened out and the target organs were identified by Bio GPS and Metascape, followed by Gene Ontology(GO) analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis of common targets. The acute drunk mouse model was established and the effects of Jingfang Granules on serum ethanol level and the expression of proteins related to the phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(Akt) signaling pathway in the liver tissue of mice were observed. A total of 187 active components of Jingfang Granules were obtained, including 47 common targets with alcoholic liver injury. GO enrichment analysis and KEGG pathway analysis showed that Jingfang Granules might play the role of relieving alcohol and protecting liver through the PI3 K-Akt signaling pathway. The drug-component-target and component-target-pathway networks revealed that the important active components of Jingfang Granules in relieving alcohol and protecting liver included quercetin, 5-O-methylvisamminol, glyasperin M, glyasperin B and hederagenin. Molecular docking showed that the active components had a good affinity with AKT1, EGFR, ESR1 and PTGS2. Experimental results showed that Jingfang Granules(15 and 10. 5 g·kg-1) could significantly reduce the content of serum ethanol in mice and up-regulate the protein expression ratios of p-PI3 K/PI3 K and p-Akt/Akt in the liver tissue. Jingfang Granules could relieve alcohol and protect liver through multi-component and multitarget, and the mechanism may be related to the activation of the PI3 K-Akt signaling pathway.

Network pharmacology and in vitro study of Fengreqing oral liquid in the intervention of wind-heat pattern.

OBJECTIVE: To investigate the underlying mechanism of the effect of Fengreqing oral liquid (, FOL) on wind-heat pattern (WHP). METHODS: In this study, we predicted the potential targets of FOL via the approach of network pharmacology and verified it by in vitro inflammation model. In the network pharmacology part, two strategies, namely the direct target search and the indirect one, were used to collect the target sets of FOL in WHP treatment. The enrichment analysis was carried out by David database and ClueGo plug-in in Cytoscape. Furthermore, the potential targets were mapped in the candidate pathways. In the verification experiment section, in vitro model of lipopolysaccharide (LPS) induced RAW 264.7 was used to confirm the predictive results in the network pharmacology part. RESULTS: Through the two screening strategies, a total of 141 non-repetitive intervention targets of FOL on WHP were obtained. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis showed that the intervention effect was mainly focused on the anti-inflammatory effect, and the Toll-like receptor signaling pathway was one of the most critical regulatory pathways. Further mapping analysis showed that phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT) signaling transfer might be the key part of regulating the concentration of inflammation mediators of FOL in the Toll-like receptor signaling pathway. In vitro experiment showed that FOL significantly reduced the levels of NO, IL-1, IL-6, and TNF-α produced by RAW264.7 induced by LPS. Further immunofluorescence found that this effect is related to the regulation of PI3K-AKT pathway activity by FOL. CONCLUSION: FOL can intervene in WHP by regulating the content of inflammatory mediators via the PI3K-AKT pathway.

Anxiolytic potency of iridoid fraction extracted from Valeriana jatamansi Jones and its mechanism: a preliminary study.

Valeriana jatamansi Jones (V. jatamansi) has been widely used for treating anxiety and its mechanism involves many aspects including GABA level. This study aimed to evaluate the anxiolytic potency of an iridoid fraction extracted from the radix and rhizomes of V. jatamansi. The iridoid fraction was extracted by using D101 resin; its major components were analysed preliminarily by thin layer chromatography, ultraviolet spectrophotometry and high-performance liquid chromatography; and its anxiolytic effects at 6 mg/kg (low-dose), 9 mg/kg (medium-dose) and 12 mg/kg (high-dose) were evaluated using the elevated plus maze test, the light-dark box test, the Vogel's drinking conflict test, and the open field drink test. Its action mechanism was investigated using the ELISA. This study provided evidence on the anxiolytic potency of the iridoid fraction from V. jatamansi and revealed its action mechanism of regulating the GABA level.