Pharmacokinetic study of five components of Fuzheng Huayu tablets in healthy human plasma.

Natural medicines play a crucial role in clinical drug applications, serving as a primary traditional Chinese medicine for the clinical treatment of liver fibrosis. Understanding the in vivo metabolic process of the Fuzheng Huayu (FZHY) formula is essential for delving into its material basis and mechanism. In recent years, there has been a growing body of research focused on the mechanisms and component analysis of FZHY. This study aimed to examine the pharmacokinetics of FZHY in healthy volunteers following oral administration. Blood samples were collected at designated time intervals after the oral intake of 9.6-g FZHY tablets. A UHPLC-Q/Exactive method was developed to assess the plasma concentrations of five components post-FZHY ingestion. The peak time for all components occurred within 10 min. The peak concentration (Cmax ) values for amygdalin, schisandrin, and schisandrin A ranged from 3.47 to 28.80 ng/mL, with corresponding AUC(0-t) values ranging from 10.63 to 103.20 ng h/mL. For schisandrin B and prunasin, Cmax values were in the range of 86.52 to 229.10 ng/mL, and their AUC(0-t) values ranged from 375.26 to 1875.54 ng h/mL, indicating significant exposure within the body. These findings demonstrate that the developed method enables rapid and accurate detection and quantification of the five components in FZHY, offering a valuable reference for its clinical study.

The molecular pathogenesis of triptolide-induced hepatotoxicity.

Triptolide (TP) is the major pharmacologically active ingredient and toxic component of Tripterygium wilfordii Hook. f. However, its clinical potential is limited by a narrow therapeutic window and multiple organ toxicity, especially hepatotoxicity. Furthermore, TP-induced hepatotoxicity shows significant inter-individual variability. Over the past few decades, research has been devoted to the study of TP-induced hepatotoxicity and its mechanism. In this review, we summarized the mechanism of TP-induced hepatotoxicity. Studies have demonstrated that TP-induced hepatotoxicity is associated with CYP450s, P-glycoprotein (P-gp), oxidative stress, excessive autophagy, apoptosis, metabolic disorders, immunity, and the gut microbiota. These new findings provide a comprehensive understanding of TP-induced hepatotoxicity and detoxification.

Antibiotic pretreatment promotes orally-administered triptolide absorption and aggravates hepatotoxicity and intestinal injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Triptolide (TP) exhibits extensive pharmacological activity, but its hepatotoxicity and intestinal injury are significant and limit its clinical use. AIM OF THE STUDY: To investigate the effect of gut microbiota disturbance after antibiotic pretreatment on TP-induced hepatotoxicity, intestinal injury and their mechanism. MATERIALS AND METHODS: We compared the characteristics of TP-induced hepatotoxicity and intestinal injury in mice with or without antibiotic pretreatment. The levels of cytokines in the serum, immunohistochemistry, and the pharmacokinetics of TP were determined. RESULT: Antibiotic pretreatment aggravates TP-induced hepatotoxicity and ileum/colon injury. TP induces hepatotoxicity in a dose-dependent manner after antibiotic pretreatment. Serum IL-1ß and IL-6 levels were increased in mice given oral TP after antibiotic pretreatment. TP can increase the expression of NLRP3 inflammasome in hepatocytes, and Oral TP after antibiotic pretreatment can significantly enhance its expression, but NLRP3 inflammasome no significant change in colon and ileum. The pharmacokinetic characteristics of TP are altered significantly by antibiotic pretreatment, as shown by a 145.87% increase in Cmax, a 155.11% increase in AUC0-t, a 155.1% increase in relative bioavailability, and a 15.44% delay in MRT. Moreover, TP causes hepatotoxicity in a time-dependent manner. CONCLUSIONS: Antibiotic pretreatment aggravates triptolide-induced hepatotoxicity and intestinal injury through elevated inflammatory response and promoted triptolide absorption.

Simultaneous determination of eight analytes of Fuzheng Huayu recipe in beagle dog plasma by UHPLC-Q/exactive Orbitrap HRMS and its application to toxicokinetics.

Fuzheng Huayu recipe (FZHY) is a Chinese patent medicine for the treatment of liver fibrosis. This study aimed to investigate the toxicokinetics of FZHY in beagle dogs after oral administration. Blood samples were collected on days 1, 15 and 28 after oral gavage of FZHY dosages of 400 or 1,200 mg/kg body weight once a day. A UHPLC-Q-Orbitrap method was developed and validated to simultaneously determine and quantify eight components of FZHY in beagle dog plasma. The times to peak concentration for eight components were18-120 min. The peak concentrations (Cmax ) of amygdalin, genistein, daidzein and 3,4-dihydroxybenzaldehyde were 1.43-43.50 ng/ml, the areas under the concentration-time curve (AUC(0-t) ) were 2.45-6,098.25 ng min/ml, and the apparent volumes of distribution (Vd ) were 0.05-131.23 × 104 ml/kg. The values of Cmax of prunasin, schisantherin A, schisandrin A and schisandrin were 7.35-1,450.73 ng/ml, the values of AUC(0-t) were 3,642.30-330,388.65 ng min/ml, and the values of Vd were 11.15-1,087.18 × 104 ml/kg. No obvious accumulation of the eight compounds was observed in beagle dogs. The results showed that the method is rapid, accurate and sensitive, and is suitable for detecting the eight analytes of FZHY. This study provides an important basis for the assessment of FZHY safety.

Systematic Evaluation of the Mechanisms of Mulberry Leaf (Morus alba Linne) Acting on Diabetes Based on Network Pharmacology and Molecular Docking.

BACKGROUND: Diabetes mellitus is one of the most common endocrine metabolic disorder- related diseases. The application of herbal medicine to control glucose levels and improve insulin action might be a useful approach in the treatment of diabetes. Mulberry leaves (ML) have been reported to exert important activities of anti-diabetic. OBJECTIVE: In this work, we aimed to explore the multi-targets and multi-pathways regulatory molecular mechanism of Mulberry leaves (ML, Morus alba Linne) acting on diabetes. METHODS: Identification of active compounds of Mulberry leaves using Traditional Chinese Medicine Systems Pharmacology (TCMSP) database was carried out. Bioactive components were screened by FAF-Drugs4 website (Free ADME-Tox Filtering Tool). The targets of bioactive components were predicted from SwissTargetPrediction website, and the diabetes related targets were screened from GeneCards database. The common targets of ML and diabetes were used for Gene Ontology (GO) and pathway enrichment analysis. The visualization networks were constructed by Cytoscape 3.7.1 software. The biological networks were constructed to analyze the mechanisms as follows: (1) compound-target network; (2) common target-compound network; (3) common targets protein interaction network; (4) compound-diabetes protein-protein interactions (ppi) network; (5) target-pathway network; and (6) compound-target-pathway network. At last, the prediction results of network pharmacology were verified by molecular docking method. RESULTS: 17 active components were obtained by TCMSP database and FAF-Drugs4 website. 51 potential targets (11 common targets and 40 associated indirect targets) were obtained and used to build the PPI network by the String database. Furthermore, the potential targets were used for GO and pathway enrichment analysis. Eight key active compounds (quercetin, Iristectorigenin A, 4- Prenylresveratrol, Moracin H, Moracin C, Isoramanone, Moracin E and Moracin D) and 8 key targets (AKT1, IGF1R, EIF2AK3, PPARG, AGTR1, PPARA, PTPN1 and PIK3R1) were obtained to play major roles in Mulberry leaf acting on diabetes. And the signal pathways involved in the mechanisms mainly include AMPK signaling pathway, PI3K-Akt signaling pathway, mTOR signaling pathway, insulin signaling pathway and insulin resistance. The molecular docking results show that the 8 key active compounds have good affinity with the key target of AKT1, and the 5 key targets (IGF1R, EIF2AK3, PPARG, PPARA and PTPN1) have better affinity than AKT1 with the key compound of quercetin. CONCLUSION: Based on network pharmacology and molecular docking, this study provided an important systematic and visualized basis for further understanding of the synergy mechanism of ML acting on diabetes.

Integrated network pharmacology and molecular docking strategy to explore the mechanism of medicinal and edible Astragali Radix-Atractylodis Macrocephalae Rhizoma acting on pneumonia via immunomodulation.

Pneumonia refers to a death-causing infection. Astragali Radix (AR) and Atractylodis Macrocephalae Rhizoma (AMR) are widely used as traditional tonic and promising edible immunomodulatory herbal medicine, but the systemic mechanism is not well understood. Therefore, a strategy based on network pharmacology and molecular docking was designed to explore the systemic mechanism of AR-AMR acting on pneumonia. After a series of bioinformatics assays, seven kernel targets were obtained, including TNF, IL6, IFNG, IL1B, IL10, IL4, and TLR9. And seven key compounds were identified as the synergy components of AR-AMR acting on pneumonia, the four key compounds belonging to AR were (3R)-3-(2-hydroxy-3,4-dimethoxyphenyl)-7-chromanol, formononetin, quercetin, and kaempferol, the three key compounds belonging to AMR were atractylone, 14-acetyl-12-senecioyl-2E, 8E, 10E-atractylentriol, and α-Amyrin. The crucial pathways were mainly related to three modules, including immune diseases, infectious disease, and organismal systems. Collectively, these observations strongly suggest that the molecular mechanisms of AR-AMR regulating pneumonia were closely related to the correlation between inflammation and immune response. PRACTICAL APPLICATIONS: Astragali radix and Atractylodis macrocephalae rhizoma can be used as "medicine-food homology" for dietary supplement. AR and AMR are widely used as a traditional tonic and promising edible immunomodulatory herbal medicine. The AR-AMR herb pairs are used for compatibility many times in the recommended prescriptions in COVID-19 develop pneumonia in China. However, the ingredients and mechanisms of AR-AMR acting on Pneumonia via immunomodulation are unclear. In this paper, bioinformatics and network biology were used to systematically explore the mechanisms of the AR-AMR herb pairs in treatment of pneumonia, and further analyze the correlation mechanism between it and COVID-19 develop pneumonia. To sum up, our study reveals the interrelationships between components, targets, and corresponding biological processes of AR-AMR acting on pneumonia. Understanding these relationships may provide guidance and theoretical basis for the further application of AR-AMR herb pairs.

Virtual Screening of Potential Anti-fatigue Mechanism of Polygonati Rhizoma Based on Network Pharmacology.

BACKGROUND AND OBJECTIVE: A large number of people are facing the danger of fatigue due to the fast-paced lifestyle. Fatigue is common in some diseases, such as cancer. The mechanism of fatigue is not definite. Traditional Chinese medicine is often used for fatigue, but the potential mechanism of Polygonati Rhizoma (PR) is still not clear. This study attempts to explore the potential anti-fatigue mechanism of Polygonati Rhizoma through virtual screening based on network pharmacology. METHODS: The candidate compounds of PR and the known targets of fatigue are obtained from multiple professional databases. PharmMapper Server is designed to identify potential targets for the candidate compounds. We developed a Herbal medicine-Compound-Disease-Target network and analyzed the interactions. Protein-protein interaction network is developed through the Cytoscape software and analyzed by topological methods. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment are carried out by DAVID Database. Finally, we develop Compound-Target-Pathway network to illustrate the anti-fatigue mechanism of PR. RESULTS: This approach identified 12 active compounds and 156 candidate targets of PR. The top 10 annotation terms for GO and KEGG were obtained by enrichment analysis with 35 key targets. The interaction between E2F1 and PI3K-AKT plays a vital role in the anti-fatigue effect of PR due to this study. CONCLUSION: This study demonstrates that PR has multi-component, multi-target and multipathway effects.

[Purification of total flavonoids from the extracts of Thesium Chinese with macroporous adsorption resin].

OBJECTIVE: To optimize the process parameters for purifying total flavonoids from Thesium chinese with D101 macroporous adsorption resin. METHODS: Purification technology of total flavonoids from the extracts of Thesium chinese was investigated by dynamic adsorption method with the transfer rates and the purity of total flavonoids as indexes. RESULTS: The optimal purifying conditions were as follows: sample concentration was 2.259 mg/mL with a speed of 1 BV/h, eluent concentration was 70% alcohol with a speed of 1BV/h. After purification, the transfer rate of total flavonoids was 94.44% and the purity of total flavonoids achieved 12.45%, which was 4 times of that by coarse extraction (2.91%). CONCLUSION: The purification technology is simple, stable and can significantly improve the contents of total flavoids in extracts.

Effects of a selective cyclooxygenase-2 inhibitor, nimesulide, on the growth of ovarian carcinoma in vivo.

New therapies against cancer are based on targeting cyclooxygenase-2 (COX-2). Whether COX-2 inhibitor therapy would be beneficial in the prevention and/or treatment of ovarian cancer still remains unclear. This study was designed to investigate whether nimesulide, a COX-2 selective inhibitor, could suppress tumor growth in implanted ovarian carcinoma mice and to explore the molecular mechanisms. Human ovarian SKOV-3 carcinoma cells xenograft-bearing mice were treated with nimesulide 62.5 mg/kg or 250 mg/kg alone i.g., daily for 21 days. Microvessel density (MVD) of ovarian carcinoma was determined with anti-CD(34) as the label. Prostaglandin E2 (PGE2) levels were also determined by ELISA. In addition, the expression of COX-2 and COX-1 at protein and mRNA levels in the control groups was also detected by immunohistochemistry and reverse-transcription polymerase chain reaction (RT-PCR). Nimesulide treatment showed a dose-dependent growth-inhibitory effect of human ovarian SKOV-3 tumors. The inhibitory rates in nimesulide 62.5 mg/kg group and 250 mg/kg group were 20.40% and 50.55% respectively, however, which is not significant statistically compared with that of control group (P > 0.05). In treatment groups, nimesulide significantly reduced intratumor PGE2 levels (all, P < 0.01). Microvessel densities in treatment groups were 61.20 +/- 1.67 (62.5 mg/kg) and 66.27 +/- 1.20 (250 mg/kg), which are significant statistically compared with that of control group (79.97 +/- 1.07) (all, P < 0.01). However, COX-1, not COX-2, mRNA, and protein levels are elevated in tumor tissues. Nimesulide decreased microvessel density is associated with the reduction of PGE2 levels but without affecting growth inhibition and the expression of COX-2. Importantly, tumor growth implanted in SKOV-3 mice was not significantly attenuated suggesting that COX-1 in ovarian carcinoma tissue also has an important role in tumor growth. These findings may implicate COX-1 as a suitable target for the treatment of ovarian cancer.

[Expressions of Cx43 and Skp2 in epithelial ovarian tumor and their clinical significances].

BACKGROUND & OBJECTIVE: Connexin 43 (Cx43), an important member of connexins family, is frequently down-regulated in neoplastic cells. It has been shown to have gap junction-dependent anti-tumor effect on various tumor cell lines. Recently, it was reported that Cx43 could inhibit tumor growth via down-regulating the expression of S-phase kinase associated protein 2 (Skp2). Skp2, a member of F-box protein family, can specifically recognize, and accelerate the ubiquitin-mediated degradation of several key regulators of G(1) phase progression. This study was to detect expressions of Cx43 and Skp2 in epithelial ovarian tumor, and to explore their correlations to tumorigenesis and development of ovarian cancer. METHODS: Expressions of Cx43 and Skp2 were examined by immunohistochemistry in 81 specimens of epithelial ovarian tumor (13 specimens of adenoma, 12 specimens of borderline adenoma, and 56 specimens of adenocarcinoma). Relationship between expression levels of Cx43 and Skp2, and association of their expression levels with clinicopathologic factors were statistically analyzed. RESULTS: Positive rates of Cx43 in ovarian adenoma, borderline adenoma, and ovarian adenocarcinoma were 84.6% (11/13), 66.7% (8/12), and 33.9% (19/56), respectivelyu expression level of Cx43 in ovarian adenocarcinoma was significantly lower than those in ovarian adenoma (P<0.01), and borderline adenoma (P<0.01). Positive rates of Skp2 in ovarian adenoma, borderline adenoma, and ovarian adenocarcinoma were 0, 0, and 46.3% (26/56), respectivelyu expression level of Skp2 in ovarian adenocarcinoma was significantly higher than those in ovarian adenoma (P<0.01), and borderline adenoma (P<0.01). Moreover, the expression levels of Cx43 and Skp2 were independent of age and histological type, but significantly associated with pathologic grade, clinical stage, and positive lymph node metastasis of ovarian adenocarcinoma. Besides, in ovarian adenocarcinoma, expression level of Cx43 was moderately inversely correlated with that of Skp2 (r=-0.48, P<0.01). CONCLUSIONS: Down-regulation of Cx43, and over-expression of Skp2 are tumor specific, and may play important roles in tumorigenesis, and development of ovarian cancer. Up-regulation of Skp2 may be related with down-regulation of Cx43 in ovarian cancer.