Unraveling a Combined Inactivation Mechanism of Cytochrome P450s by Genipin, the Major Reactive Aglycone Derived from Gardeniae Fructus.

Genipin (GP) is the reactive aglycone of geniposide, the main component of traditional Chinese medicine Gardeniae Fructus (GF). The covalent binding of GP to cellular proteins is suspected to be responsible for GF-induced hepatotoxicity and inhibits drug-metabolizing enzyme activity, although the mechanisms remain to be clarified. In this study, the mechanisms of GP-induced human hepatic P450 inactivation were systemically investigated. Results showed that GP inhibited all tested P450 isoforms via distinct mechanisms. CYP2C19 was directly and irreversibly inactivated without time dependency. CYP1A2, CYP2C9, CYP2D6, and CYP3A4 T (testosterone as substrate) showed time-dependent and mixed-type inactivation, while CYP2B6, CYP2C8, and CYP3A4 M (midazolam as substrate) showed time-dependent and irreversible inactivation. For CYP3A4 inactivation, the kinact/KI values in the presence or absence of NADPH were 0.26 or 0.16 min-1 mM-1 for the M site and 0.62 or 0.27 min-1 mM-1 for the T site. Ketoconazole and glutathione (GSH) both attenuated CYP3A4 inactivation, suggesting an active site occupation- and reactive metabolite-mediated inactivation mechanism. Moreover, the in vitro and in vivo formation of a P450-dependent GP-S-GSH conjugate indicated the involvement of metabolic activation and thiol residues binding in GP-induced enzyme inactivation. Lastly, molecular docking analysis simulated potential binding sites and modes of GP association with CYP2C19 and CYP3A4. We propose that direct covalent binding and metabolic activation mediate GP-induced P450 inactivation and alert readers to potential risk factors for GP-related clinical drug-drug interactions.

Draft genome sequence of a new carotenoid-producing strain Brevibacterium sp. XU54, isolated from radioactive soil in Xinjiang, China.

Some species of the genus Brevibacterium are orange bacteria involved in cheese ripening, synthesis of odoriferous compounds, and carotenoids with aromatic end groups. Here, we report the genome sequence of Brevibacterium sp. XU54, isolated from radioactive soil in Xinjiang, China. The genome of XU54 consists of 4,899,099 base pairs with a GC content of 62.2%. The genome sequence was annotated with 4453 genes, encoding 4260 proteins, 13 rRNAs, and 49 tRNAs. 16S rRNA BLAST and comparative genomic analysis both indicated that XU54 may be a new species of Brevibacterium. In addition, compared to the type strains, some enzymes related to sulfur metabolism showed a low similarity of 66.85, 79.53 and 14.61%, respectively. The carotenoids biosynthesis gene cluster was identified and analyzed according to the genomic data, which revealed relatively low identity (5-85%) with existing strains. The optimum conditions for its growth and carotenoid production were then discussed. The whole-genome sequence of Brevibacterium sp. XU54 will be beneficial for utilizing these newly identified genes in carotenoid biosynthesis and regulation of sulfur metabolism pathway to promote the production of novel carotenoids and other structurally diverse compounds through combinatorial biosynthesis, which facilitates cheese ripening and coloration. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03366-1.

Gut microbiota affects the efficacy of Danggui Buxue Tang by affecting plasma concentration of active ingredients.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui Buxue Tang (DBT) is a traditional Chinese medicine, which has the function of supporting Qi and enriching blood. Antibiotics can cause Gut microbiota disorder and affect efficacy of DBT. AIM OF THE STUDY: Explore the manner in which Gut microbiota affects the efficacy of Danggui Buxue Tang. MATERIALS AND METHODS: In this study, antibiotics were used to destroy gut microbiota. The changes of DBT efficacy were detected to verify the effect of gut microbiota on DBT efficacy. The changes of gut microbiota was detected using 16S rRNA sequencing, and UPLC-MS/MS was used to analyze the plasma concentration of active ingredients. Correlation analysis was used to establish the relationship between gut microbiota, blood components and drug efficacy, and to explore the role of gut microbiota in the efficacy of DBT. RESULTS: The results showed that the efficacy in the DBT group was significantly improved compared with the control group (p<0.05). Compared with DBT group, the efficacy in antibiotic DBT treatment (ABXDBT) group was significantly reduced, 194 plasma metabolites and 18 DBT blood components were significantly altered in ABXDBT group, and 11 DBT blood components such as caffeic acid and formononetin were significantly decreased. Correlation analysis showed that 6 DBT blood components were related with the decrease of efficacy. Network pharmacology analysis showed that the above 6 DBT blood components participated in the hematopoietic regulation through PI3K-Akt and HIF-1 signaling pathways. Correlation analysis showed that Bacteroides and other intestinal bacteria were related to the absorption of DBT active ingredients. The drug metabolic pathway of gut microbiota was significantly decreased after antibiotic treatment (p = 0.033). CONCLUSIONS: Gut microbiota such as Bacteroides affects the efficacy of DBT by affecting the metabolism and absorption of DBT active ingredients such as caffeic acid and formononetin.

Danggui Buxue Tang restores antibiotic-induced metabolic disorders by remodeling the gut microbiota.

ETHNOPHARMACOLOGICAL RELEVANCE: Danggui Buxue Tang (DBT) has been used to promote hematopoiesis and relieve myelosuppression in China. Antibiotics can cause myelosuppression through gut microbiota disorders. AIM OF THE STUDY: This study aims to explore the way of DBT to alleviate the metabolic disorder caused by antibiotics. MATERIALS AND METHODS: In this study, 16S rRNA sequencing was used to detect the change of gut microbiota, metabolomics to analyze the change of metabolites. Correlation analysis was used to establishment the correlation between gut microbiota and metabolites. PICRUST 2 was used to predict the function of gut microbiota. RESULTS: Results showed that eighty-two genera of gut microbiota were affected by antibiotic, while twelve were significantly restored after DBT. Seventy-four potential metabolites were significantly different from the antibiotics and DBT. We found significant recovery by the Bacteroides and Rikenellaceae RC9 after DBT. The metabolic pathways influenced by the antibiotic treatment included primary and secondary bile biosynthesis, etc. The metabolic pathways that could be restored after DBT included the primary and secondary bile acid biosynthesis pathway, etc. Through correlation analysis, we found a correlation between the Bacteroides, Rikenellaceae_RC9_gut_group and other potential differential metabolisms such as those of taurodeoxycholic acid, N-phenylacetyl glycine, etc. The functional prediction showed that the biosynthesis of primary bile acid, secondary bile acid was significantly affected. CONCLUSIONS: DBT can restore the gut and reverse the metabolic disorder caused by antibiotics through Bacteroides, and it provides a new medical idea regarding the gut microbiota balance.

The traditional uses, phytochemistry, pharmacology and toxicology of Cinnamomi ramulus: a review.

OBJECTIVES: Cinnamomi ramulus (called Guizhi in Chinese) is a traditional medicine used to treat gastrointestinal dysfunction, cancer, arthritis, osteoporosis, spleen deficiency, Alzheimer's disease and obesity. This review aimed to provide a systematic summary on the geographical distribution, botany, traditional application, phytochemistry, pharmacology, pharmacokinetics, toxicology and other aspects of Cinnamomi ramulus. KEY FINDING: So far, more than 121 chemical compounds have been isolated from Cinnamomi ramulus, including volatile oil, organic acids, triterpenoid saponins, coumarins, tannins, flavonoids and flavonoid glycosides, steroids and polysaccharides. This paper reviews the pharmacological effects of Cinnamomi ramulus on antibacterial, anti-inflammatory, antiviral, antitumour, antipyretic and analgesic, antidiabetic and antiplatelet aggregation effects. Furthermore, the present review also indicates that Cinnamomi ramulus has the potential to develop into drugs for treating various diseases with high efficacy and low toxicity. SUMMARY: The convictive evidence from modern pharmacology research supports the traditional application of Cinnamomi ramulus. However, further studies on the structure-activity relationship of some of the isolated compounds may improve their biological potency. More toxicological studies will also contribute to the progress of clinical trial studies.

Enhanced fluoride adsorption of aluminum humate and its resistance on fluoride accumulation in tea leaves.

Tea trees (Camellia sinensis) can take in fluorine from soil and the content of fluorine in tea increases with maturity, leading to high content of fluoride in tea leaves and tea products. Long-term consumption of high fluoride tea products could result in chronic fluoride intoxication. Confining the fluoride in the earth with absorbents to reduce the fluoride accumulation of the tea trees during the growth period which could radically control the fluoride level in tea product. Humic acid (HA), a kind of organic matter in the earth was used as raw material to prepare adsorbent aluminum humate (HAA) by aluminum modification. The HAA absorbent presented excellent absorption performance to the fluoride in a wide pH range (4-10), and the maximum adsorptive capacity can reach to 62.5 mg/g. The absorption isotherm demonstrated the adsorption of fluoride was the monomolecular adsorption and the absorption was in accordance with the pseudo-second order kinetic equation. Fluoride content in real soil solution decreased significantly by 53.03% by using the HAA absorbent. The utilization of HAA adsorbent in the culture and field plots experiments also obviously adsorb the soluble fluoride in solution and soil, which could significantly suppress the fluoride accumulation in tea leaves. In September, the fluoride accumulation in tea leaves has been reduced 74.29% in the field plots experiments.

[Screening of Active Fractions from Huanglian Jiedu Decoction against Primary Neuron Injury after Oxygen-Glucose Deprivation].

OBJECTIVE: To observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability. RESULTS: RP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01). CONCLUSIONS: HJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Huanglian-Jie-Du-Tang extract protects against chronic brain injury after focal cerebral ischemia via hypoxia-inducible-factor-1α-regulated vascular endothelial growth factor signaling in mice.

Huanglian-Jie-Du-Tang (HJDT) is a traditional Chinese herbal formula which is widely used clinically. In this study, we investigated the effects of an aqueous (HJDTaq) and an ethanolic (HJDTet) extract of HJDT on chronic brain injury after focal cerebral ischemia in mice. The ischemia was induced by occlusion of the right middle cerebral artery for 30 min. HJDTaq (4 g/kg) and HJDTet (200, 400, 800 mg/kg) were orally administered for 21 d from day 7 before ischemia to day 14 after ischemia. The survival rate decreased to less than 50% at 35 d after ischemia. HJDTet at 400 mg/kg increased the survival rate. HJDTaq (4 g/kg) and HJDTet (400, 800 mg/kg) significantly attenuated the neurological dysfunction, brain atrophy and infarct volume after ischemia. There were few cells positive for CD31, hypoxia-inducible-factor-1α (HIF-1α), vascular endothelial growth factor (VEGF) and Flk-1 in the sham control. After ischemia, the number increased. HJDTaq (4 g/kg) and HJDTet (400 or 800 mg/kg) further increased the numbers of CD31, HIF-1α, VEGF and Flk-1-positive cells in the ischemic hemisphere. We conclude that HJDTaq and HJDTet have neuroprotective effects on chronic brain injury after focal cerebral ischemia and lead to accelerated angiogenesis by HIF-1α-regulated VEGF signaling.

[Comparison of volatile oils of cultivated Houttaynia cordata populations with wild].

OBJECTIVE: To provide scientific proofs for the exploitation, utilization, and normalized cultivation of Houttuynia cordata. METHOD: The underground parts of 17 wild H. cordata populations from different valleys and altitudes of mountain. Emei were transplanted to the same growth conditions. After one year's cultivation, volatile oil was obtained by steam distillation from the aerial part of the materials. The chemical constituents were separated and identified by GC-MS, and the relative content of each constituent was determined by area normalization. RESULT: Totally, 31 chemical components were identified, 19 components could be detected in all materials. The t-test results indicated that the contents of alpha-pinene and D-limonene were extremely significantly higher than that in the wild populations, and the contents of camphene and 2-undecanone were also significantly higher than that in the wild populations. And the reverse was found in the content of trans-beta-ocimene. All these 31 components could be divided into 9 chemical compositions, and 7 chemical compositions could be detected in the wild and cultivated. RSD values of 5 chemical compositions in wild populations were higher than that in the cultivated, except for the contents of diterpenyl alcohols and diterpenyl aldehydes. These five chemical compositions accounted for 84.05% and 90.12% of the whole volatile oils in the wild and cultivated conditions, respectively. CONCLUSION: The components and contents of the volatile oils between wild and cultivated were different. The volatile oils polymorphism decrease distinctively as all the wild populations of H. cordata were transplanted to the uniform environmental conditions.

[Effects of angustmycin on callus occurrence and proliferation of Panax notogingseng in vitro].

In order to explore quick, efficient inducement and proliferation of Panax notogingseng callus, the stem sections were taken as explants in the following 5 groups of medium: 1. MS + 2,4-D 2 mg/L (CK), 2. MS + 2,4-D 2 mg/L + N6-BA 2 mg/L, 3. MS + 2,4-D 2 mg/L + KT 2 mg/L, 4. MS + 2,4-D 2 mg/L + ZT 2 mg/L, 5. MS + 2,4-D 2 mg/L + LFS 2 mg/L. The results showed: 1. LFS (Lingfasu, a new kind of CTK) was able to promote the callus formation earlier 1-2 week than CK and to raise the rate of induced callus as high as 81%, higher 30% than that on other 4 mediums; 2. On medium 5, the fresh weight of callus increased 360.2% after being cultured 40 d, on other 4 mediums increased only 13.4%-21.8%. At the same time, 1 g callus cultured 40 d could obtain dry material 81.5 mg on medium 5, but on other 4 mediums only 21.5-25.9 mg; 3. The callus cultured on medium 5. continual generation to generation could live as long as more than 3 years, on other 4 mediums the callus hardened and aged quickly.