Dandelion Root Extract Induces Intracellular Ca2+ Increases in HEK293 Cells

Gerbino, Andrea; Russo, Daniela; Colella, Matilde; Procino, Giuseppe; Svelto, María; Milella, Luigi; Carmosino, Monica

doi:10.3390/ijms19041112

Cited by 16 publications

(22 citation statements)

References 54 publications

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“…The effect of DRE on ClC-Ka is mediated by the activation of PKC pathway. While dissecting the effect of DRE on Ca 2+ signaling, we demonstrated that DRE-induced Ca 2+ release from the ER is a consequence of the activation of the Gαq/PLC/IP3 pathway 18 suggesting the activation of the conventional PKC, referred downstream in this signaling cascade as follow: PLC splits phosphatidylinositol into IP3 and diacylglycerol (DAG), the latter involved in the activation of the membrane-bound PKC together with intracellular Ca 2+ 29 . To evaluate whether the effect of DRE on ClC-Ka was dependent to the activation of conventional PKC pathway, we performed DRE experiments in the presence of either the Ca 2+ chelator BAPTA or PKC inhibitor Calphostin C. As shown in Figure 3A, the inhibitory effect on ClC-Ka activity recorded after 20 min of exposure to 400 µg/mL DRE was abolished at each time point when the cells were treated with BAPTA.…”

Section: Dre Inhibits Also the Activity Of Clc-kb While Other Renal Tmentioning

confidence: 98%

“…For all the functional experiments we used a dose of DRE (400 µg/mL) that we previously characterized and tested in terms of maximal tolerated concentration and ability to mobilize intracellular Ca 2+ and activate phospholipase C (PLC) in HEK293 cells 18 . Figure 1A (Upper panel) reports representative Cl − currents recorded from the same ClC-Ka/barttin expressing HEK293 cell, at the start of the whole-cell recording, i.e.…”

Section: Dre Time-dependently Inhibits the CL − Current In Hek293 Celmentioning

confidence: 99%

“…In a previous study, we found that non-toxic doses of a hydroethanolic Dandelion Root Extract (DRE) mobilize intracellular Ca 2+ and activate PLC in HEK293 cells 18 . Interestingly, when extracts of dandelion were administered by gastric gavage to rats, a significant diuretic effect, paralleled by a weight loss, was obtained 19 .…”

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confidence: 98%

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Role of PKC in the Regulation of the Human Kidney Chloride Channel ClC-Ka

Gerbino

Zio

Russo

et al. 2020

Sci Rep

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The physiological role of the renal ClC-Ka/ClC-K1 channels is to confer a high Cl- permeability to the thin Ascending Limb of Henle (tAL), which in turn is essential for establishing the high osmolarity of the renal medulla that drives water reabsorption from collecting ducts. Here, we investigated by whole-cell patch-clamp measurements on HEK293 cells co-expressing ClC-Ka (tagged with GFP) and the accessory subunit barttin (tagged with m-Cherry) the effect of a natural diuretic extract from roots of Dandelion (DRE), and other compounds activating PKC, such as ATP, on ClC-Ka activity and its membrane localization. Treatment with 400 µg/ml DRE significantly inhibited Cl- currents time-dependently within several minutes. Of note, the same effect on Cl- currents was obtained upon treatment with 100 µM ATP. Pretreatment of cells with either the intracellular Ca2+ chelator BAPTA-AM (30 μM) or the PKC inhibitor Calphostin C (100 nM) reduced the inhibitory effect of DRE. Conversely, 1 µM of phorbol meristate acetate (PMA), a specific PKC activator, mimicked the inhibitory effect of DRE on ClC-Ka. Finally, we found that pretreatment with 30 µM Heclin, an E3 ubiquitin ligase inhibitor, did not revert DRE-induced Cl- current inhibition. In agreement with this, live-cell confocal analysis showed that DRE treatment did not induce ClC-Ka internalization. In conclusion, we demonstrate for the first time that the activity of ClC-Ka in renal cells could be significantly inhibited by the activation of PKC elicited by classical maneuvers, such as activation of purinergic receptors, or by exposure to herbal extracts that activates a PKC-dependent pathway. Overall, we provide both new information regarding the regulation of ClC-Ka and a proof-of-concept study for the use of DRE as new diuretic.

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Section: Dre Inhibits Also the Activity Of Clc-kb While Other Renal Tmentioning

confidence: 98%

Section: Dre Time-dependently Inhibits the CL − Current In Hek293 Celmentioning

confidence: 99%

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Role of PKC in the Regulation of the Human Kidney Chloride Channel ClC-Ka

Gerbino

Zio

Russo

et al. 2020

Sci Rep

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“…Some specific components from plant extracts also have the ability to bind to protein domains, leading to modification or inhibition of enzymatic activities. It has been reported that dandelion root extracts dose-dependently increase intracellular Ca 2+ levels in HEK293 cells, partially by stimulating sarco/endoplasmic reticulum Ca 2+ -ATPase (SERCA) [37]. Chatterjee et al observed a rapid activation of caspase-8 enzyme in A375 cells when treated with dandelion root extracts for 30 min, pointing to the direct effects of dandelion root extracts on enzymatic activity but not gene transcription [38].…”

Section: Discussionmentioning

confidence: 99%

Reversal of heavy metal-induced antibiotic resistance by dandelion root extracts and taraxasterol

Yang¹,

Zhang

2020

Journal of Medical Microbiology

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Introduction. Metal exposure is an important factor for inducing antibiotic resistance in bacteria. Dandelion extracts have been used for centuries in traditional Chinese and Native American medicine. Aim. We assessed the effects of dandelion water extracts and taraxasterol on heavy metal-induced antibiotic resistance in Escherichia coli as well as the underlying mechanisms. Methodology. Dandelion extracts were obtained through 4 h of boiling in distilled water. Bacterial growth was monitored with a spectrophotometer. Biochemical assays were performed to assess the activities and gene transcriptions of β-lactamase and acetyltransferase. Oxidative stress was determined using an oxidation-sensitive probe, H2DCFDA. Results. The present study demonstrated that higher concentrations of nickel (>5 µg ml−1), cadmium (>0.1 µg ml−1), arsenic (>0.1 µg ml−1) and copper (>5 µg ml−1) significantly inhibited the growth of E. coli . Lower concentrations of nickel (0.5 µg ml−1), cadmium (0.05 µg ml−1) and arsenic (0.05 µg ml−1) had no effect on bacterial growth, but helped the bacteria become resistant to two antibiotics, kanamycin and ampicillin. The addition of dandelion root extracts and taraxasterol significantly reversed the antibiotic resistance induced by these heavy metals. The supplements of antibiotics and cadmium generated synergistic effects on the activities of β-lactamase and acetyltransferase (two antibiotic resistance-related proteins), which were significantly blocked by either dandelion root extract or taraxasterol. In contrast, oxidative stress was not involved in the preventative roles of dandelion root extracts and taraxasterol in heavy metal-induced antibiotic resistance. Conclusion. This study suggests that heavy metals induce bacterial antibiotic resistance and dandelion root extracts and taraxasterol could be used to help reverse bacterial resistance to antibiotics.

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“…Its leaves are used in salads, and its roots are used as coffee 8 . It has lately been determined that some components of the plant are effective as diuretic, laxative, biliary deterrent, although the plant has usually been used as a cure for liver and gall disturbances since the ancient times 9 . Studies on the antioxidant 10 , anti-inflammatory 11 , antidiabetic 12 , antiulcer 13 , anti-rheumatic 14 , and anticancer 15 properties of T. officinale have been reported in literature.…”

Section: Introductionmentioning

confidence: 99%

Comparative Analysis of the Selected Secondary Metabolites Extracted from Wild and Micropropagated Taraxacum officinale

2018

jcbsc

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Taraxacum officinale, commonly named dandelion, is an edible plant distributed worldwide, belonging to the Asteraceae family. Due to the metabolites it contains, T. officinale has been used for medicinal purposes since the ancient times. Studies on metabolite production using plants with the methods of plant tissue culture acquire more importance every day. In this study, the metabolite contents of the Taraxacum officinale plants, naturally grown in the wild and clonally propagated in tissue culture conditions, were investigated. The micropropagation study of T. officinale was carried out in Murashige and Skoog (MS) medium including 2 mg L-1 benzyl amino purine (BAP) and 2 mg L-1 indole butyric acid (IBA). For metabolite studies, dried and grounded samples were stirred in aqueous methanol solution over three hours at room temperature. The plant extract was dried and then separated into fractions with C18 reversed-phase column chromatography. The metabolites of the samples were analyzed with Q-TOF LC-MS in negative ion mode in which more than 10 metabolites were characterized from both natural and cloned T. officinale. The presence of the same metabolites in the plant extracts showed that the micropropagation of T. officinale can be potentially used as a new protocol for the production of beneficial secondary metabolites for pharmaceutical and supplemental food industries.

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Dandelion Root Extract Induces Intracellular Ca2+ Increases in HEK293 Cells

Cited by 16 publications

References 54 publications

Role of PKC in the Regulation of the Human Kidney Chloride Channel ClC-Ka

Role of PKC in the Regulation of the Human Kidney Chloride Channel ClC-Ka

Reversal of heavy metal-induced antibiotic resistance by dandelion root extracts and taraxasterol

Comparative Analysis of the Selected Secondary Metabolites Extracted from Wild and Micropropagated Taraxacum officinale

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