Trophic Transfer without Biomagnification of Cadmium in a Soybean-Dodder Parasitic System

Cadmium (Cd) is among the most available and most toxic heavy metals taken up by plants from soil. Compared to the classic plant-animal food chains, the host-parasitic plant food chains have, thus far, been largely overlooked in the studies of Cd trophic transfer. To investigate the pattern of Cd transfer during the infection of parasitic plants on Cd-contaminated hosts, we conducted a controlled experiment that grew soybeans parasitized by Chinese dodders (Cuscuta chinensis) in soil with different levels of Cd treatment, and examined the concentration, accumulation, allocation and transfer coefficients of Cd within this parasitic system. Results showed that among all components, dodders accounted for more than 40% biomass of the whole system but had the lowest Cd concentration and accumulated the least amount of Cd. The transfer coefficient of Cd between soybean stems and dodders was much lower than 1, and was also significantly lower than that between soybean stems and soybean leaves. All these features were continuously strengthened with the increase of Cd treatment levels. The results suggested no evidence of Cd biomagnification in dodders parasitizing Cd-contaminated hosts, and implied that the Cd transfer from hosts to dodders may be a selective process.

Study on Active Components of Cuscuta chinensis Promoting Neural Stem Cell Proliferation: Bioassay-Guided Fractionation

Neural stem cells (NSCs) exist in the central nervous system of adult animals and capable of self-replication. NSCs have two basic functions, namely the proliferation ability and the potential for multi-directional differentiation. In this study, based on the bioassay-guided fractionation, we aim to screen active components in Cuscuta chinensis to promote the proliferation of NSCs. CCK-8 assays were used as an active detection method to track the active components. On the basis of isolating active fraction and monomer compounds, the structures of these were identified by LC-MS and (1H, 13C) NMR. Moreover, active components were verified by pharmacodynamics and network pharmacology. The system solvent extraction method combined with the traditional isolation method were used to ensure that the fraction TSZE-EA-G6 of Cuscuta chinensis exhibited the highest activity. Seven chemical components were identified from the TSZE-EA-G6 fraction by UPLC-QE-Orbitrap-MS technology, which were 4-O-p-coumarinic acid, chlorogenic acid, 5-O-p-coumarinic acid, hyperoside, astragalin, isochlorogenic acid C, and quercetin-3-O-galactose-7-O-glucoside. Using different chromatographic techniques, five compounds were isolated in TSZE-EA-G6 and identified as kaempferol, kaempferol-3-O-glucoside (astragalin), quercetin-3-O-galactoside (hyperoside), chlorogenic acid, and sucrose. The activity study of these five compounds showed that the proliferation rate of kaempferol had the highest effects; at a certain concentration (25 μg/mL, 3.12 μg/mL), the proliferation rate could reach 87.44% and 59.59%, respectively. Furthermore, research results using network pharmacology techniques verified that kaempferol had an activity of promoting NSCs proliferation and the activity of flavonoid aglycones might be greater than that of flavonoid glycosides. In conclusion, this research shows that kaempferol is the active component in Cuscuta chinensis to promote the proliferation of NSCs.

Effect of total flavonoids of Cuscuta chinensis Lam. (Convolvulaceae) on oxidative stress injury in mouse testis and epididymis, and on serum levels of reproductive hormones in oligoasthenospermia mice model

Purpose: To investigate the effect of total flavonoids of Cuscuta chinensis (TFCC) on oxidative stress injury in testis and epididymis, and serum levels of reproductive hormones in an oligoasthenospermia (OAS) mice model. Methods: Thirty male Wistar mice were randomly assigned to three groups of 10 mice each: control group, OAS group and TFCC group. With the exception of control group, OAS was orally induced in the mice with ornidazole. The TFCC group received TFCC. Reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) were determined. Serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone were also measured. Results: The levels of ROS and MDA in the testis and epididymis significantly increased in OAS group, when compared to control mice (p < 0.05). However, TFCC administration significantly reduced their levels in these tissues (p < 0.05). In contrast, SOD activity significantly decreased in the testis and epididymis of mice in OAS group, relative to control group, but increased significantly after TFCC exposure (p < 0.05). Serum FSH and LH were markedly elevated in OAS group, but treatment with TFCC significantly reduced the levels of these hormones (p < 0.05). Conclusion: These results suggest that TFCC effectively improves sperm quality and reduces oxidative damage in testis and epididymis of mice with oligoasthenospermia via a mechanism involving the regulation of serum levels of reproductive hormones. Thus, TFCC may be useful in the treatment of oligoasthenospermia.

Toxicity Effects of Chinese Herbal ‘Five-seeds’ Formulation on Human Kidney HEK-293 and Chang Liver Cells

‘Five-seeds’ formulation (Lycium barbarum, Cuscuta chinensis Lam, Rubus idaeus, Schisandra chinensis, Plantago asiatica) have been used by Chinese medicine practitioner for the treatment of male infertility for a long time. The present study aims to investigate the potential toxicity effects of the individual herb used in ‘five-seeds’ formulation and as a concoction on human HEK293 and Chang liver cells, using cell viability MTT assay and SubG1 flow cytometry analysis. Percentage of cell viability from samples (1, 25, 50, 75, 100 mg/mL) decreased as the concentration increased. Half maximal inhibitory concentration (IC50) values indicated that Cuscuta chinensis, Rubus idaeus and Schisandra chinensis are among the most toxic herb samples on HEK293 cells, with the IC50 values less than 22 mg/mL. On the other hand, Rubus idaeus (IC50 value 20.1 mg/mL) and Schisandra chinensis (IC50 value 17.8 mg/mL) were found to have the highest toxicity effect on Chang liver cells. Plantago asiatica and Lycium barbarum (IC50 more than 67 mg/mL) are the least toxic herbs tested on both human HEK293 kidney and Chang liver cells. ‘Five-seeds’ herbal formulation had IC50 values of 33 mg/mL and 38.5 mg/mL on HEK293 and Change liver cells respectively, suggesting that ‘five seeds’ formulation has modulated the toxicity effect of the mixed herbs used in this formula. Apoptotic cells in SubG1 phase were also found to be significantly low for HEK293 (13.08%) and Chang liver cells (10.17%) with the treatment of 25 mg/mL of ‘five seeds’ formulation. Toxicity effect of five-seeds’ herbal formulation has seemingly been modulated when all the five herbs mixed as a concoction, and potentially non-toxic as a concoction to HEK293 kidney and Chang liver cells. However, more investigations should be done to draw a solid conclusion.

Traditional Chinese Medicine Drynariae Rhizoma and Cuscuta Chinensis Suppress Osteoarthritis by Quercetin-AKT1 and Luteolin-IL6/VEGFA Direct Binding

Background: Drynaria Fortunei and Cuscuta Chinensis are among the most used traditional Chinese medicine herbal prescriptions and have a significant therapeutic effect on osteoarthritis. However, the purpose of this study intends to elaborate the molecular mechanism of action through network pharmacology. The active ingredients of TCM and the potential targets for the treatment of osteoarthritis were selected through the TCMSP, OMIM and Genecards. Results: The 27 components and 85/117 targets of Drynaria Fortunei and/or Cuscuta Chinensis were identified for osteoarthritis. Pharmacological and PPI network analysis identified top 3 active components (kaempferol, luteolin, and quercetin) and core proteins (IL6, AKT1, and VEGFA). GO and KEGG analysis identified the top 3 functions (cytokine and cell/nuclear receptor) and pathways (PI3K-Akt, TNF and IL-17). Molecular docking showed strong binding ability between quercetin-AKT1 and luteolin-IL6/VEGFA. Interaction analysis mapped the quercetin-AKT1 and luteolin-IL6/VEGFA binding to specific hydrogen and hydrophobic bonds. Conclusions: The main active components, common target proteins, functional activities, and signaling pathways of TCM Drynaria Fortunei and Cuscuta Chinensis for the treatment of osteoarthritis were identified by Network pharmacology. We found, for the first time, that drynariae rhizoma and cuscuta chinensis suppress osteoarthritis by quercetin-AKT1/IL6 and luteolin-VEGFA direct binding. Our findings have significant implication for our understanding of the molecular mechanism of action in the treatment of osteoarthritis and future development of osteoarthritis treatment using quercetin and luteolin.

PHYTOCONSTITUENTS FROM THE ROOTS OF CUSCUTA CHINENSIS

Cuscuta chinensis, a parasitic vine, was used in several traditional medicine systems, and it demonstrated a wider range of pharmacological activities in various diseases. The chemical components from C. chinensis consist mainly of flavonoids, steroids, volatile constituents, lignans, alkaloids, and polysaccharides. In view of its wider pharmacological properties, the authors have attempted to re-investigate the whole plant for its phytoconstituents and reported flavones salvigenin, chrisimaritin for the first time apart from the earlier reported quercetin and triterpenoid betulinic acid.