Clerodendranthus Spicatus Research Articles

Clerodendranthus spicatus (Thunb.) Water Extracts Reduce Lipid Accumulation and Oxidative Stress in the Caenorhabditis elegans.

Clerodendranthus spicatus (Thunb.) (Kidney tea) is a very distinctive ethnic herbal medicine in China. Its leaves are widely used as a healthy tea. Many previous studies have demonstrated its various longevity-promoting effects; however, the safety and specific health-promoting effects of Clerodendranthus spicatus (C. spicatus) as a dietary supplement remain unclear. In order to understand the effect of C. spicatus on the longevity of Caenorhabditis elegans (C. elegans), we evaluated its role in C. elegans; C. spicatus water extracts (CSw) were analyzed for the major components and the effects on C. elegans were investigated from physiological and biochemical to molecular levels; CSw contain significant phenolic components (primarily rosmarinic acid and eugenolinic acid) and flavonoids (primarily quercetin and isorhamnetin) and can increase the lifespan of C. elegans. Further investigations showed that CSw modulate stress resistance and lipid metabolism through influencing DAF-16/FoxO (DAF-16), Heat shock factor 1 (HSF-1), and Nuclear Hormone Receptor-49 (NHR-49) signalling pathways; CSw can improve the antioxidant and hypolipidemic activity of C. elegans and prolong the lifespan of C. elegans (with the best effect at low concentrations). Therefore, the recommended daily use of C. spicatus should be considered when consuming it as a healthy tea on a daily basis.

Anti-hyperuricemia effect of Clerodendranthus spicatus: a molecular biology study combined with metabolomics

Hyperuricemia (HUA), a metabolic disease caused by excessive production or decreased excretion of uric acid (UA), has been reported to be closely associated with a variety of UA transporters. Clerodendranthus spicatus (C. spicatus) is an herbal widely used in China for the treatment of HUA. However, the mechanism has not been clarified. Here, the rat model of HUA was induced via 10% fructose. The levels of biochemical indicators, including UA, xanthine oxidase (XOD), adenosine deaminase (ADA), blood urea nitrogen (BUN), and creatinine (Cre), were measured. Western blotting was applied to explore its effect on renal UA transporters, such as urate transporter1 (URAT1), glucose transporter 9 (GLUT9), and ATP-binding cassette super-family G member 2 (ABCG2). Furthermore, the effect of C. spicatus on plasma metabolites was identified by metabolomics. Our results showed that C. spicatus could significantly reduce the serum levels of UA, XOD, ADA and Cre, and improve the renal pathological changes in HUA rats. Meanwhile, C. spicatus significantly inhibited the expression of URAT1 and GLUT9, while increased the expression of ABCG2 in a dose-dependent manner. Metabolomics showed that 13 components, including 1-Palmitoyl-2-Arachidonoyl-sn-glycero-3-PE, Tyr-Leu and N-cis-15-Tetracosenoyl-C18-sphingosine, were identified as potential biomarkers for the UA-lowering effect of C. spicatus. In addition, pathway enrichment analysis revealed that arginine biosynthesis, biosynthesis of amino acids, pyrimidine metabolism and other metabolic pathways might be involved in the protection of C. spicatus against HUA. This study is the first to explore the mechanism of anti-HUA of C. spicatus through molecular biology and metabolomics analysis, which provides new ideas for the treatment of HUA.

Physical properties, phenolic profile and antioxidant capacity of Java tea (Clerodendranthus spicatus) stems as affected by steam explosion treatment

Java tea (Clerodendranthus spicatus) has been favored for its various health benefits and abundance of phenolic substances. Steam explosion (SE) treatment was performed in the pretreatment of Java tea stems and the physical properties, phenolic profile and antioxidant capacity were investigated. Extraction kinetics study showed that the phenolics yields of Java tea stems treated at 2.4 MPa for 10 min reached the maximum in 40 min, which was approximately 3 times the yields of raw stems in 180 min. The antioxidant activities of the extracts of Java tea stems were also significantly increased after SE treatment (P < 0.05). In addition, 19 phenolics were detected in Java tea stems by HPLC/QTOF‐MS/MS, and rosmarinic acid was found to be hydrolyzed to danshensu during the SE process. SE could be an efficient pretreatment technology to improve the extraction rates of phenolics and conversions of their high-value hydrolyzed products, which could facilitate further research of Java tea products.

Phenylboronic Acid and Amino Bifunctional Modified Adsorbent for Quickly Separating Phenolic Acids from Crude Extract of Clerodendranthus spicatus and Evaluation of Their Antioxidant and Hypoglycemic Activities.

A novel phenylboronic acid and amino bifunctional modified silica gel (SiO2-NH2-FPBA) was prepared, which was 30-80 μm, had a pore size of 8.69 nm, a specific surface area of 206.89 m2/g, was stable at low temperature, and contained 0.4793 mmol/g of the phenylboronic acid group and 1.6377 mmol/g of the amino group. It was used to develop a rapid separation method for phenolic acids. The results showed that it could adsorb 93.64 mg/g caffeic acid, 89.35 mg/g protocatechuic acid and 79.66 mg/g gallic acid. The adsorption process was consistent with the pseudo-second-order model (R2 > 0.99), and fitted the Langmuir isotherm model well (R2 > 0.99). CH3COOH could effectively desorb phenolic acids (>90%) and did not destroy their structures. When SiO2-NH2-FPBA was added to crude extract of Clerodendranthus spicatus, 93.24% of the phenolic acids could be captured, and twenty-two kinds of phenolic acids were identified by Q Exactive HF LC-MS. Furthermore, the isolated phenolic acids from Clerodendranthus spicatus possessed great DPPH, ABTS, and hydroxyl radicals scavenging activities and ferric reducing power. They also demonstrated effective inhibition of α-amylase and α-glucosidase activities (IC50 = 110.63 ± 3.67 μg/mL and 64.76 ± 0.30 μg/mL, respectively). The findings indicate that SiO2-NH2-FPBA has significant potential in practical applications of separating active constituents from natural resources.

Target immobilization on glass microfiber membranes as a label-free strategy for hit identification.

The discovery of novel chemical entities targeting G protein-coupled receptors (GPCRs) is usually guided by their receptor affinity. However, traditional affinity assay methods and hit identification procedures are usually laborious and expensive. In this work, the type-2 vasopressin receptor (V2R) was chosen as a prototypical GPCR. Membrane fragments from cells highly expressing SNAP-V2R were immobilized on the surface of a glass microfiber (GMF) coated with O6-benzylguanine (BG). This was achieved by transferring the benzyl group of BG to the active site of the SNAP-tag through a nucleophilic substitution reaction. As a result, a biofilm called SNAP-V2R@GMF-BG was produced that showed good specificity and stability. The adsorption ratio for each V2R ligand treated with SNAP-V2R@GMF-BG was determined by HPLC and exhibited a good linear correlation with the Ki value determined by displacement assays. Furthermore, a Ki prediction assay was performed by comparing the data with that generated by a homogeneous time-resolved fluorescence (HTRF) assay. SNAP-V2R@GMF-BG was also used to screen hit compounds from natural products. After SNAP-V2R@GMF-BG was incubated with the total extract, the ligand that binds to V2R could be separated and subjected to LC‒MS analysis for identification. Baicalein was screened from Clerodendranthus spicatus and verified as a potential V2R antagonist. This V2R-immobilized GMF platform can help determine the affinity of V2R-binding hit compounds and screen the compounds efficiently and accurately.

Clerodendranthus spicatus inhibits epithelial–mesenchymal transition of renal tubular cells through the NF-κB/Snail signalling pathway in hyperuricaemia nephropathy

Context Clerodendranthus spicatus Thunb. (Labiatae) (CS), a perennial traditional Chinese medicinal herb that can reduce serum uric acid (sUA) levels and ameliorate renal function is widely used to treat hyperuricaemic nephropathy (HN). Objective To investigate the molecular mechanism of action of CS in HN treatment using in vivo and in vitro experiments. Materials and methods Sprague-Dawley rats were randomly divided into control, HN, CS and positive control allopurinol groups. The HN group was intraperitoneally injected with 750 mg/kg oxonic acid potassium (OA), whereas the CS group was injected with OA along with a gavage of CS (low dose 3.125 g/kg, high dose 6.25 g/kg) for five weeks. For in vitro studies, uric acid-treated HK2 cells were used to verify the therapeutic mechanism of CS in HN. Results HN rats exhibit pathological phenotypes of elevated sUA levels and renal injury. CS significantly improved these symptoms and sUA (p < 0.05) and blood urea nitrogen (p < 0.01) levels, and dramatically improved renal tubular injury in HN rats. The IC50 value of UA (uric acid) in HK2 cells was 826.32 ± 3.55 μg/mL; however, 120 ng/mL CS had no significant cytotoxicity on HK2 cells. In vivo and in vitro studies showed that CS inhibited NF-κB phosphorylation and inhibited α-smooth muscle actin (α-SMA) and vimentin expression while increasing E-cadherin expression, suggesting that CS inhibited the fibrotic process in renal cells, thus protecting renal function. Discussion and conclusions These findings provide a fundamental understanding of the application of CS in HN treatment to better guide clinical interventions.

Steam explosion improves extractability, antioxidant activity and α-glucosidase inhibitory activity of the constituents of Java tea (Clerodendranthus spicatus)

In this work, the steam explosion (SE) technology was exploited to extract valuable phytochemicals from Java tea (Clerodendranthus spicatus). Modification effects of SE on apparent morphology and physicochemical properties of Java tea were investigated to confirm that SE destroyed the cell wall structure and enhanced pore space, which facilitated the dissolution of ingredients. After SE pretreatment, the contents of phenolics, triterpenes, and sterols of Java tea extracts were maximally increased by 40.68%, 25.69%, and 40.11% in the leaves group, and increased by 147.09%, 204.50%, and 115.24% in the stems group, respectively. The antioxidant and α-glucosidase inhibition capacity of extracts were also obviously enhanced. Multivariate analysis indicated that phenolics and triterpenes were the main active substances in the extracts, and the leaves treated at 1.0 MPa and stems treated at 2.4 MPa showed higher comprehensive score. This study provided a promising method for Java tea pretreatment for improving extractability and bioactivity.

Kidney tea ameliorates hyperuricemia in mice via altering gut microbiota and restoring metabolic profile

Clerodendranthus spicatus (Thunb.) C. Y. Wu, also known as kidney tea (KT), has been widely employed in kidney protection in Chinese Medicine. It has been reported that KT can lower uric acid (UA) and mitigate gout, while the mechanism remains to be elucidated. Given the close relationship between hyperuricemia (HUA), intestinal flora and host metabolism, this study aimed to explore the mechanism by which KT lowers UA from the perspective of the fecal microbiome and metabolome. Initially, mice were intraperitoneally injected with potassium oxonate to induce the HUA model. The results showed that KT markedly reduced the serum level of UA and impaired renal damage in HUA mice. Subsequently, the result of 16S rRNA gene sequencing analysis indicated that KT administration appeared a significant improvement in the structure of the intestinal flora, especially increased the abundances of Roseburia and Enterorhabdus, while decreased the abundances of Ileibacterium and UBA1819. Moreover, the levels of differential metabolites (including twenty-five in feces and eight in serum) identified by untargeted metabolomics returned to normal after KT intervention. Taken together, the mechanism of KT in alleviating HUA is related to the regulation of the intestinal flora and the remodeling of metabolic disorders, which will lay a theoretical foundation for KT as a UA-lowering drug.

Studies on the key constituents and the related mechanisms of Clerodendranthus spicatus in the treatment of diabetes based on network pharmacology

Ethnopharmacological relevanceClerodendranthus spicatus is a traditional Chinese medicine and has been used to treat diabetes and some kidney diseases for a long history. Aim of the studyThe research aimed to study the active constituents, the potential targets and the related mechanisms of C. spicatus in the treatment of diabetes through network pharmacology method and verify the antidiabetic activity by molecular biology experiments. Materials and methodsA comprehensive network pharmacology strategy was used to predict the key active constituents, the key targets and the related mechanisms and pathways of C. spicatus in the treatment of diabetes. The strategy mainly included screening and predicting potential active constituents and targets by network construction, GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis. Based on the predicted results, C. spicatus was extracted by ultrasonic method with 50% ethanol and enriched by using macroporous resin. The compounds with potential antidiabetic effects were separated through silica-gel column chromatography and HPLC (high performance liquid chromatography), and then identified by MS (mass spectrum) and NMR (nuclear magnetic resonance). The C. spicatus extract and isolated compounds were tested by in-vitro and cell experiments to verify their antidiabetic activities, including antioxidant activities, inhibition activities on α-glucosidase and α-amylase, the influence on glucose uptake in cell experiments and the Western blot of PI3K and Akt expression levels. ResultsA total of 18 active constituents and 16 key targets of C. spicatus in the treatment of diabetes were screened out through network pharmacology method. Phenolic acids might be the main target compounds for the next research. After extraction, enrichment and separation, the phenolic acids-enriched fraction of C. spicatus and four phenolic acid compounds (helisterculin C, salvianolic acid B, orthosiphoic acid E and ethyl caffeate) were obtained. Among them, salvianolic acid B was isolated from C. spicatus for the first time and orthosiphoic acid E was isolated from natural products for the first time. In experiment verification, the crude extract of C. spicatus, the phenolic acids-enriched fraction and the four compounds all showed antidiabetic potentials. The phenolic acids in C. spicatus had antioxidant activities, inhibitory activities on α-amylase and α-glucosidase and promoted glucose uptake in L6 cells through PI3K/Akt signaling pathway. ConclusionsThis study showed that C. spicatus had antidiabetic activities with the mechanism of the mode of multi-compounds acting on multi-targets and multi-pathways. The main active phenolic acid compounds were also identified. It provided theoretical basis for further development and utilization of C. spicatus.

The protective effects of Clerodendranthus spicatus (Thunb.) C. Y. Wu extract on oxidative stress induced by 2,2'-azo (2-methylpropamidine) dihydrochloride in HL-1 mouse cardiomyocytes.

To investigate the protective effects of Clerodendranthus spicatus (Thunb.) C. Y. Wu extract (CSTE) on oxidative stress injury in HL-1 mouse cardiomyocytes induced by 2,2'-azo (2-methylpropamidine) dihydrochloride (AAPH, 1 mmol/L), HL-1 cells were co-cultured with different concentrations (10–100 μg/mL) of the CSTE for 24 h. A cell damage model was established by continuously culturing the cells in Dulbecco's Modified Eagle Medium plus AAPH for 4 h. Cell survival rates were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay, and by measuring intracellular malondialdehyde (MDA) content. MDA and total reactive oxygen species (ROS) levels were determined by thiobarbituric acid colorimetry and the 2',7'-dihydrodichlorofluorescent sodium yellow diacetate probe, respectively. Apoptosis was measured by flow cytometry. The intracellular catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione s-transferase (GST), γ-glutamylcysteine synthetase (γ-GCS), and glutathione (GSH) contents were determined by colorimetric methods. CSTE content was determined by high performance liquid chromatography. The CSTE pretreatment improved survival rates in damaged HL-1 cells, reduced total intracellular ROS and MDA levels, and reduced apoptosis. The CSTE also increased the activities of the antioxidant enzymes (CAT, SOD, GSH-Px, and GST), as well as the γ-GCS and GSH levels in damaged cells. Real-time fluorescence quantitative polymerase chain reaction analysis indicated that the CSTE upregulated CAT, SOD1, and GSH-Px mRNA expression levels. Additionally, the CSTE reduced MDA and ROS levels in HL-1 cells by improving the endogenous antioxidant system; thus, alleviating the oxidative stress damage caused by AAPH. Our compositional analyses revealed that the CSTE contained caffeic acid, isoquercetin, rosmarinic acid, luteolin, and baicalin. The CSTE demonstrates antioxidant and protective effects in myocardial cells.

The complete chloroplast genome sequence of Clerodendranthus spicatus, a medicinal plant for preventing and treating kidney diseases from Lamiaceae family

Clerodendranthus spicatus (Thunb.) C. Y. Wu ex H. W. Li is one of the most important medicines for the treatment of nephrology in the southeast regionsof China. To understand the taxonomic classification of Clerodendranthus species and identify species discrimination markers, we sequenced and characterized its chloroplast genome in the current study. Total genomic DNA were isolated from dried leaves of C. spicatus and sequenced using an Illumina sequencing platform. The data were assembled and annotated by the NOVOPlasty software and CpGAVAS2 web service. The complete chloroplast genome of C. spicatus was 152,155bp, including a large single-copy region of 83,098bp, a small single-copy region of 17,665bp, and a pair of inverted repeat regions of 25,696bp. The Isoleucine codons are the most abundant, accounting for 4.17% of all codons. The codons of AUG, UUA, and AGA demonstrated a high degree of usage bias. Twenty-eight simple sequence repeats, thirty-six tandem repeats, and forty interspersed repeats were identified. The distribution of the specific rps19, ycf1, rpl2, trnH, psbA genes were analyzed. Analysis of the genetic distance of the intergenic spacer regions shows that ndhG-ndhI, accD-psaI, rps15-ycf1, rpl20-clpP, ccsA-ndhD regions have high K2p values. Phylogenetic analysis showed that C. spicatu is closely related to two Lamiaceaespecies, Tectona grandis, and Glechoma longituba. In this study, we sequenced and characterized the chloroplast genome of C. spicatus. Phylogenomic analysis has identified species closely related to C. spicatus, which represent potential candidates for the development of drugs improving renal functions.

Effect of Clerodendranthus spicatus (Thunb.) C. Y. Wu on the exercise ability of D-galactose-induced oxidative aging mice

The Clerodendranthus spicatus (Thunb.) C. Y. Wu extract (CSTCYWE) was administered to D-galactose-induced aging mice to investigate the effect and mechanism of CSTCYWE on exercise endurance in aging mice. The effect of CSTCYWE on the improvement of exercise capacity in aging mice was evaluated by detecting serum biochemical indicators, pathological changes in tissues, and expression by qPCR. The composition of CSTCYWE was analyzed by HPLC. The experiment showed that CSTCYWE increased the running time and the exhaustive swimming time of aging mice to improve their exercise endurance. CSTCYWE also reduced the levels of BUN, BLA, and MDA and increased the levels of HG, MG, SOD, and GSH-Px in the serum of aging mice. Pathological observation found that CSTCYWE alleviated the liver and kidney tissue lesions and damage caused by aging. Meanwhile, CSTCYWE could down-regulate the relative mRNA expressions of nNOS, iNOS, TNF-α, and syncytin-1 and up-regulate the relative mRNA expressions of Cu/Zn-SOD, Mn-SOD, and CAT in the mouse liver tissue and skeletal muscle tissue. The results of component analysis showed that CSTCYWE contained caffeic acid, hypericin, isoquercetin, dihydroquercetin, rosmarinic acid, baicalin, luteolin, and baicalein. These active ingredients enable CSTCYWE to act on aging mice and improve their exercise capacity.

Studies on the Key Constituents and the Related Mechanisms of Clerodendranthus Spicatus in the Treatment of Diabetes Based on Network Pharmacology

Studies on the Key Constituents and the Related Mechanisms of Clerodendranthus Spicatus in the Treatment of Diabetes Based on Network Pharmacology

Bioactivity-guided separation of potential \u03b1-glycosidase inhibitor from clerodendranthus spicatus based on HSCCC coupled with molecular docking

Clerodendranthus Spicatus is a traditional Dais medi-edible plant and it has been proven to have good blood glucose-lowering efficacy. However, the material basis of Clerodendranthus Spicatus has not been clarified yet and therefore needs to be determined. In this paper, the effective ingredients of this medicine were purified by high-speed counter-current chromatography. Alongside, their potential hypoglycemic activity was determined by α-glucosidase inhibitory activities in vitro and molecular docking. Finally, five compounds were purified and identified as 2-caffeoyl-L-tartaric acid (1), N-(E)-caffeoyldopamine (2), rosmarinc acid (3), methyl rosmarinate (4), 6,7,8,3′,4′-Pentamethoxyflavone (5). Examination of α-glucosidase inhibitory activity in vitro showed that 2-caffeoyl-L-tartaric acid and rosmarinic acid had a higher inhibitory activity than acarbose. Molecular docking indicated that the affinity energy of the identified compounds ranged from − 7.6 to − 8.6 kcal/mol, a more desirable result than acarbose (− 6.6 kcal/mol). Particularly, rosmarinc acid with the lowest affinity energy of − 8.6 kcal/mol was wrapped with 6 hydrogen bonds. Overall, α-glucosidase inhibitory activities and molecular docking suggested that rosmarinc acid was likely to be a promising hypoglycemic drug.

Bioassay-guided isolation of anti-inflammatory diterpenoids with highly oxygenated substituents from kidney tea (Clerodendranthus spicatus).

The whole plant of Clerodendranthus spicatus (Thunb.) is one of popular functional food in south of China, named as "kidney tea" and used to ameliorate renal inflammation. In order to verify this potential function and explore the accurate compounds responsible for inflammation, the ethanol extract, fractions, and subfractions of this plant were prepared to evaluate anti-inflammation effect on xylene-induced acute inflammatory mice model, and the results indicated that two subfractions from EtOAc fraction show potential activities. Subsequent bioassay-guided isolation of the bioactive subfractions led to isolation of 25 compounds. Among them, compounds 2, 4, 5, 9-11, 13, 16, 17, and 20-22 inhibited the productions of pro-inflammation factors TNF-α, IL-1β, and IL-8 in lipopolysaccharide (LPS) -induced renal epithelia (HK-2) cells, respectively. Further anti-inflammation evaluation in vivo indicated that the major bioactive compounds 1, 2, 5-7, 17, 21, and 22 from C. spicatus were even better than aspirin. PRACTICAL APPLICATIONS: C. spicatus as a healthy tea has been available in the Chinese market and as a medicine for various disorders such as nephritis, rheumatism, inflammation, gout, and diabetes. Previous pharmacological investigation of the plant revealed the potential anti-inflammatory activities, but the material basis of anti-inflammatory activity remains to be elucidated. In our study, the anti-inflammatory fractions and compounds were obtained by the bioassay-guide isolation and the results showed that the highly oxygenated diterpenoids were major anti-inflammatory compounds, in which 1, 2, 5-7, 17, 21, and 22 were even better than aspirin. This information supported kidney tea as a functional food for treatment of renal inflammation reasonably and may add a new dimension to biological activity of this plant in the field of agriculture as a functional food were cultivated.

"Kidney Tea" and Its Bioactive Secondary Metabolites for Treatment of Gout.

Clerodendranthus spicatus, popularly known as "kidney tea" in China, is consumed traditionally as a functional food for treatment of renal inflammation, dysuria, and gout. We evaluated the effects of C. spicatus on gout by assessing activities of antihyperuricemia, anti-gouty arthritis, and analgesia in vivo, and the results indicated that the ethyl acetate fraction shows potential activities. Subsequent phytochemical investigation of this fraction led to the isolation of 32 compounds, consisting of 20 diterpenoids (including the new orthosiphonones E and F), 2 triterpenoids, 6 flavonoids, 2 lignanoids, and 2 phenolic acid derivatives. Pharmacological investigation of the pure compounds in the cellular model of hyperuricemia indicated that 12 compounds could promote the excretion of uric acid at 10 μg/mL, and compounds 3, 4, 5, and 21 had better effects than that of benzbromarone, a famous uricosuric drug. Furthermore, compounds 4, 6, 7, 9, 14, 15, 23, 26, and 31 showed significant anti-gouty arthritis activity in monosodium urate (MSU)-induced joint swelling at the dose of 50 mg/kg, while compounds 4, 5, 7, 9, and 26 exhibited significant inhibition of pain induced by acetic acid. Our findings provided scientific justification to support the traditional application of "kidney tea" for treating gout and suggested its good application prospects in the future.

Can intercropping with the Chinese medicinal herbs change the water use of the aged rubber trees?

Intercropping the medicine herbs (i.e., Amomum villosum and Clerodendranthus spicatus) with rubber tree in Xishuangbanna prefecture of southwestern China is regarded as a promising solution to reduce the negative hydrological effects which were caused by the monocultural cultivation of rubber tree and improve the sustainability of aged rubber plantation. However, the water-use characteristics of rubber tree in these agroforestry systems (AFSs) were still rarely reported, and our current understanding of the below-ground competition in such tree and herb AFSs is also insufficient. Therefore, we adopted the techniques of stable isotopes to study the water use efficiency (WUE; through leaf δ13C) and the water-absorbing patterns (through δ2H and δ18O) of the aged rubber trees and the intercropped herbs for the investigation of their water competition, and we also measured the contents of their leaf carbon (C), nitrogen (N) and phosphorus (P) to check the competition effects on the nutrient status of rubber tree. As the results of Bayesian mixing model (MixSIAR) suggested, the aged rubber trees in the rubber monoculture still absorbed the surface and shallow soil water (0–15 cm depths) mainly, same as the intercropped herbs in these AFSs, but rubber trees in the AFSs prefer to absorb the middle and deep soil water (below 15 cm depths). Such phenomenon verified that the water-absorbing patterns of these aged rubber trees were still flexible. However, the leaf δ13C of the rubber trees in these AFSs indicated their WUE was not improved through the intercropping, and thus the soil water content decreased in the AFSs. In short, intercropping the shallow roots medicine herbs (i.e., A. villosum and C. spicatus) can help the aged rubber trees use deeper soil water, and thus help the rubber tree and intercrops form complementary water-absorbing patterns in the pronounced dry season, but there is no improvement in their WUE and the soil water conditions. In addition, the decreased leaf N of rubber trees in the AFSs suggests that the N absorption of rubber trees was limited. Therefore, intercropping the medicine herbs with rubber tree still need more consideration on the design of such AFSs, especially in terms of the soil water conservation and management.

A new eudesmane type sesquiterpene from cultivated Clerodendranthus spicatus in Hainan

Six compounds were isolated from the aerial part of cultivated Clerodendranthus spicatus in Hainan with various chromatographic techniques,and their structures were determined as:1-dehydroxy-1-oxo-rupestrinol(1),N-trans-feruloyltyramine(2),methyl 3,4-dihydroxyphenyllactate(3),caffein acid(4),methyl caffeate(5) and ethyl caffeate(6),via analysis of physicochemical properties and spectroscopic evidence.Compound 1 was a new compound,while compounds 2 and 3 were isolated from C.spicatus for the first time.Biological activity results showed that compounds 2-4 exhibited α-glucosidase inhibitory activity with different inhibition ratio.

Metabolomic study of the mechanism of Dai medicine &amp;lt;italic&amp;gt;Orthosiphon stamineus&amp;lt;/italic&amp;gt; on &amp;ldquo;Jiajie&amp;rdquo;

Herb genomics constitutes the metabolomics of Chinese medicine. Dai medicine is one of the “four national medicines” in China, which are based on a theoretically perfect system, and “Jiajie” has a unique medical theory. Clerodendranthus spicatus (Thunb.) C. Y. Wu (Lamiaceae) ( Orthosiphon stamineus Thunb., OS), is one of the Dai “antidote” medicines and has significant health care applications and “detoxification” properties in the clinical setting. This study used HPLC-MS to detect changes in metabolites in rat urine and serum after the oral administration of OS extract. The orthogonal projections to latent structures discriminant analysis (OPLS-DA) method was employed to analyze metabolites of rat urine and serum in OS and control groups, and 39 differential metabolites in urine and 26 in serum were screened out. Finally, related metabolic pathways were analyzed using the MetaboAnalyst website. The oral administration of OS extract mainly affected the citrate cycle (TCA cycle), taurine and hypotaurine metabolism, tyrosine metabolism, and sphingolipid metabolism pathways. Our findings suggested that oral OS extract administration may improve basic metabolite levels and immunity in humans by improving the detoxification ability of the body through an “antidote” mechanism. This study provides a scientific basis for elucidating the mechanism underlying “detoxification” by OS. Moreover, this study offers a point of reference for the clinical application of OS, development of “Jiajie” species in the Dai medicine, and the clarification of “preventive treatment of disease” theory in traditional Chinese medicine.

Analysis of Xanthine Oxidase Inhibitors from Clerodendranthus spicatus with Xanthine Oxidase Immobilized Silica Coated Fe3O4 Nanoparticles

In this study, xanthine oxidase immobilized silica coated Fe3O4 nanoparticles (Fe3O4@SiO2-XO) were successfully prepared and characterized by transmission electron microscope, X-ray powder diffraction, Fourier transform infrared spectroscopy and vibrating sample magnetometer. The average diameter of the Fe3O4 nanoparticles was about 300 nm to 350 nm with a shell thickness of 60 nm. The maximum saturation magnetization of the Fe3O4@SiO2-XO nanoparticles was 44.9 emu/g, which ensured the separation from the medium within one minute by using an ordinary magnet. A xanthine oxidase (XO) inhibitor screening method using Fe3O4@SiO2-XO nanoparticles was established and utilized in the extract of Clerodendranthus spicatus. Under the optimized conditions, two compounds were screened out and identified as gardenin B and eupatorin. The half maximal inhibitory concentration (IC50) values of these two compounds were 1.488 μg/mL and 11.197 μg/mL, respectively. The interactions between these two compounds and XO were investigated by the fluorescence spectroscopic method. The results suggested that the quenching effects of gardenin B and eupatorin were due to a static quenching mechanism. Furthermore, gardenin B showed stronger binding capacity than that of eupatorin. In conclusion, this screening method exhibited efficiency and reusability in screening, identification and analysis of enzyme inhibitors from complex mixtures.