scholarly journals Pulchinenosides from Pulsatilla Chinensis Increase P-Glycoprotein Activity and Induce P-Glycoprotein Expression

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Yali Liu ◽  
Ling Zhang ◽  
Shaofeng Wei ◽  
Jinyang Cai ◽  
Zhenzhong Zang ◽  
...  
Keyword(s):  
Membrane Vesicles ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Intestinal Epithelial ◽  
Glycoprotein P ◽  
P Glycoprotein ◽  
Human Intestinal Epithelial Cells ◽  
Pulsatilla Chinensis ◽  
Dependent Transport ◽  
Cytotoxic Studies

Five pulchinenosides (pulchinenoside B3, pulchinenoside BD, pulchinenoside B7, pulchinenoside B10, and pulchinenoside B11) isolated from Pulsatilla chinensis (Bge) Regel saponins extract exhibited strong antitumor activities but poor gastrointestinal absorption properties. The enteric induction of P-glycoprotein (P-gp) is understood to restrict the oral bioavailability of some pharmaceutical compounds and lead to adverse drug reactions. Therefore, the present investigation was intended to delineate the impacts of pulchinenosides on cellular P-gp function and expression using Sf9 membrane vesicles and LS180 cells as a surrogate of human intestinal epithelial cells. Preliminary cytotoxic studies showed that 10 μM was an acceptable concentration for cytotoxicity and antiproliferation studies for all pulchinenosides using the alamarBlue assay. The cell cycle of LS180 cells detected by flow cytometry was not significantly influenced after 48 hours of coincubation with 10 μM of pulchinenosides. In the presence of pulchinenosides, the ATP-dependent transport of N-methyl-quinidine mediated by P-glycoprotein was stimulated significantly. The upregulation of P-glycoprotein and mRNA levels was found by Western blot and real-time PCR analysis in LS180 cells. Parallel changes indicate that all pulchinenosides are exposed to pulchinenosides-mediated transcriptional regulation. In conclusion, pulchinenosides could induce P-glycoprotein expression and directly increase its functional activity.

Hepatocyte Nuclear Factor 1Î' Upregulates P-Glycoprotein Expression in Human Intestinal Epithelial Cells

2017 ◽  
Vol 152 (5) ◽  
pp. S66-S67 ◽  
Author(s):  
Shubha Priyamvada ◽  
Mumtaz Anwar ◽  
Anoop Kumar ◽  
Arivarasu Natarajan Anbazhagan ◽  
Ravinder K. Gill ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Nuclear Factor ◽  
Intestinal Epithelial ◽  
Hepatocyte Nuclear Factor ◽  
P Glycoprotein ◽  
Human Intestinal Epithelial Cells

scholarly journals The Antibiotic Bacitracin Protects Human Intestinal Epithelial Cells and Stem Cell-Derived Intestinal Organoids from Clostridium difficile Toxin TcdB

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Ziyu Zhu ◽  
Leonie Schnell ◽  
Bastian Müller ◽  
Martin Müller ◽  
Panagiotis Papatheodorou ◽  
...  
Keyword(s):  
Stem Cell ◽  
Epithelial Cells ◽  
Inhibitory Effect ◽  
Target Cells ◽  
Intestinal Epithelial ◽  
Intestinal Organoids ◽  
Human Intestinal Epithelial Cells ◽  
Protein Toxins ◽  
Dependent Transport

Bacitracin is an established antibiotic for local application and inhibits the cell wall synthesis of Gram-positive bacteria. Recently, we discovered a completely different mode of action of bacitracin and reported that this drug protects human cells from intoxication by a variety of medically relevant bacterial protein toxins including CDT, the binary actin ADP-ribosylating toxin of Clostridium (C.) difficile. Bacitracin prevents the transport of CDT into the cytosol of target cells, most likely by inhibiting the transport function of the binding subunit of this toxin. Here, we tested the effect of bacitracin towards TcdB, a major virulence factor of C. difficile contributing to severe C. difficile-associated diseases (CDAD) including pseudomembranous colitis. Bacitracin protected stem cell-derived human intestinal organoids as well as human gut epithelial cells from intoxication with TcdB. Moreover, it prevented the TcdB-induced disruption of epithelia formed by gut epithelium cells in vitro and maintained the barrier function as detected by measuring transepithelial electrical resistance (TEER). In the presence of bacitracin, TcdB was not able reach its substrate Rac1 in the cytosol of human epithelial cells, most likely because its pH-dependent transport across cell membranes into the cytosol is decreased by bacitracin. In conclusion, in addition to its direct antibiotic activity against C. difficile and its inhibitory effect towards the toxin CDT, bacitracin neutralizes the exotoxin TcdB of this important pathogenic bacterium.

ATP-dependent transport of beta-estradiol 17-(beta-D-glucuronide) in rat canalicular membrane vesicles

1996 ◽  
Vol 271 (5) ◽  
pp. G791-G798
Author(s):  
M. Vore ◽  
T. Hoffman ◽  
M. Gosland
Keyword(s):  
Bile Acid ◽  
Transport System ◽  
Organic Anion ◽  
Membrane Vesicles ◽  
Anion Transport ◽  
Organic Anion Transport ◽  
Canalicular Membrane ◽  
P Glycoprotein ◽  
Anion Transport System ◽  
Dependent Transport

The ATP-dependent transport of beta-estradiol 17-(beta-D-glucuronide) (E217G), a cholestatic metabolite of estradiol, was investigated in rat liver canalicular membrane vesicles. ATP-dependent transport was dependent on time and temperature and occurred into an osmotically sensitive space; kinetic analysis indicated a saturable transport system (Michaelis-Menten constant value, 75 microM; maximum transport rate, 598 pmol.min-1.mg protein-1). The steroid conjugates estradiol glucuronide, estriol 3-glucuronide, estriol 16 alpha-glucuronide, testosterone glucuronide, and the three-sulfate conjugate of 17G were effective inhibitors of transport. Bromosulfophthalein, S-(2,4-dinitrophenyl)glutathione, and glutathione disulfide, all substrates of the canalicular ATP-dependent non-bile acid organic anion transport system, were also effective inhibitors, whereas taurocholate had no effect on transport. Conversely, E217G inhibited the ATP-dependent transport of S-(2,4-dinitrophenyl)glutathione. Daunorubicin, vinblastine, etoposide, cyclosporin, and PSC-833, substrates/modulators of P-glycoprotein, were also potent inhibitors of E217G transport, and E217G competitively inhibited the ATP-dependent transport of daunorubicin. C219, a monoclonal antibody against P-glycoprotein, inhibited ATP-dependent transport of E217G and daunorubicin but not of taurocholate or S-(2,4-dinitrophenyl)glutathione. These data indicate that E217G is substrate of both the non-bile acid organic anion transport system and P-glycoprotein but not of the ATP-dependent bile acid transport system in canalicular membranes.

scholarly journals Angiotensin II Inhibits P‐glycoprotein in Human Intestinal Epithelial cells

2012 ◽  
Vol 26 (S1) ◽  
Author(s):  
Seema Saksena ◽  
Anoop Kumar ◽  
Vikas Soni ◽  
Waddah A Alrefai ◽  
Pradeep K Dudeja ◽  
...  
Keyword(s):  
Angiotensin Ii ◽  
Epithelial Cells ◽  
Intestinal Epithelial Cells ◽  
Intestinal Epithelial ◽  
P Glycoprotein ◽  
Human Intestinal Epithelial Cells

209. P-glycoprotein limits activation of the glucocorticoid receptor in placental BeWo cells

2005 ◽  
Vol 17 (9) ◽  
pp. 80
Author(s):  
P. J. Mark ◽  
B. J. Waddell
Keyword(s):  
Glucocorticoid Receptor ◽  
Cell Line ◽  
Pcr Analysis ◽  
Target Cells ◽  
Glycoprotein P ◽  
Bewo Cells ◽  
P Glycoprotein ◽  
Growth Inhibitory ◽  
Induced Apoptosis ◽  
And Function

Inadequate placental growth and function are key determinants of fetal growth retardation. Glucocorticoids potently inhibit fetal and placental growth via activation of the glucocorticoid receptor (GR). Placental and fetal glucocorticoid exposure is minimised by the ‘placental glucocorticoid barrier’, which consists primarily of placental 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2) converting maternal glucocorticoids to inactive metabolites. Recent studies in the rodent brain show that P-glycoprotein (P-gp) is also an important physiological regulator of glucocorticoid access to the GR in target cells. Therefore, we hypothesised that placental P-gp may serve to exclude maternal glucocorticoids from the placenta and fetus, and thereby augment the barrier. We have used the placental choriocarcinoma cell line BeWo, and MDR-BeWo, a daughter cell line virally transduced with P-gp, to assess whether P-gp regulates access of dexamethasone to the GR. Quantitative PCR analysis showed that MDR-BeWo cells express ~10-fold higher levels of P-gp mRNA than BeWo cells. Syncytialisation of BeWo and MDR-BeWo cells with 20μM forskolin also increased P-gp mRNA by ~7-fold in each cell line. The elevated P-gp expression in MDR-BeWo cells resulted in a reduced activation of the GR with 1μM dexamethasone by ~50% (P<0.001) in comparison to BeWo cells. Accordingly, dexamethasone-induced apoptosis was reduced in MDR-BeWo cells, as indicated by a lack of induction of cleaved caspase 3 protein. Additionally, the P-glycoprotein inhibitor cyclosporin A (10μM) did not increase the level of dexamethasone-induced GR activation in the low P-gp expressing BeWo cells, but potentiated GR activation by ~2-fold in the MDR-BeWo cells, to a level comparable to that in BeWo cells. These data support the hypothesis that P-glycoprotein contributes to the placental glucocorticoid barrier. Thus, 11β-HSD2 and P-glycoprotein are likely to act in unison to reduce fetal and placental exposure to maternal glucocorticoids and minimise their growth inhibitory actions.

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