Cannabis (Cannabis sativa)
Cannabidiol changes P-gp and BCRP expression in trophoblast cell lines.
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Objectives.Marijuana is the most commonly used illicit drug during pregnancy. Due
to high lipophilicity, cannabinoids can easily penetrate physiological barriers like
the human placenta and jeopardize the developing fetus.We evaluated the impact
of cannabidiol (CBD), a major non-psychoactive cannabinoid, on P-glycoprotein
(P-gp) and Breast Cancer Resistance Protein (BCRP) expression, and P-gp function
in a placental model, BeWo and Jar choriocarcinoma cell lines (using P-gp induced
MCF7 cells (MCF7/P-gp) for comparison).
Study design. Following the establishment of the basal expression of these transporters
in the membrane fraction of all three cell lines, P-gp and BCRP protein
and mRNA levels were determined following chronic (24–72 h) exposure to CBD,
by Western Blot and qPCR. CBD impact on P-gp efflux function was examined
by uptake of specific P-gp fluorescent substrates (calcein-AM, DiOC2(3) and rhodamine123(
rh123)). Cyclosporine A (CsA) served as a positive control.
Results. Chronic exposure to CBD resulted in significant changes in the protein and
mRNA levels of both transporters. While P-gp was down-regulated, BCRP levels
were up-regulated in the choriocarcinoma cell lines. CBD had a remarkably different
influence on P-gp and BCRP expression in MCF7/P-gp cells, demonstrating that
these are cell type specific effects. P-gp dependent efflux (of calcein, DiOC2(3) and
rh123) was inhibited upon short-termexposure to CBD.
Conclusions. Our study shows that CBD might alter P-gp and BCRP expression in
the human placenta, and inhibit P-gp efflux function.We conclude that marijuana
use during pregnancy may reduce placental protective functions and change its
morphological and physiological characteristics.
1 . Inhibition of P-gp by Cannabidiol 10µM (id=NPDI-cqaAMA)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system BeWo
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
60 min
7.4
N/A
10 µM
2 . Inhibition of P-gp by Cannabidiol 10 µM (id=NPDI-IoxesQ)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system BeWo
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
60 min
7.4
10 µM
3 . Inhibition of P-gp by Cannabidiol 25 µM (id=NPDI-oZGcUg)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system BeWo
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
60 min
7.4
25 µM
4 . Inhibition of P-gp by Cannabidiol 25 µM (id=NPDI-mLsAbQ)
In Vitro Transporter Inhibition Experiment
Inhibition was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system MCF7/P-gp
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
60 min
7.4
25 µM
1 . Induction of BCRP transporter (mRNA) by Cannabidiol (id=NPDI-spUvqw)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system Jar
Results
figure 5
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
mRNA expression
Cannabis
24h, 48h, 72h
qRT-PCR, 2 CT method
2 . Induction of P-gp transporter (protein) by Cannabidiol (id=NPDI-mbNW9A)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system MCF7/P-gp
Results
figure 4
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM and 15 µM
Cannabis
48h, 72h
computerized image analysis system
3 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-Aasqdw)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system BeWo
Results
FIGURE 2
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM and 15 µM
Cannabis
72h
computerized image analysis system
4 . Induction (mRNA) of BCRP transporter with Cannabidiol (id=NPDI-jHdxpA)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system BeWo
Results
figure 5
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
mRNA expression
Cannabis
24h, 48h, 72h
qRT-PCR, 211CT method
5 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-Ix-KKg)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system Jar
Results
FIGURE 3
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM and 15 µM
Cannabis
72h
computerized image analysis system
6 . Induction of BCRP transporter (protein) by Cannabidiol (id=NPDI-olqNRw)
In Vitro Transporter Induction Experiment
Induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system Jar
Results
figure 3
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM
Cannabis
48h, 72h
computerized image analysis system
7 . Non-induction of BCRP transporter (protein) by Cannabiodiol (id=NPDI-PaO28A)
In Vitro Transporter Induction Experiment
Non-induction was detected. Cutoff used —
N/A
- BCRP (ABCG2)
Cell system MCF7/P-gp
Results
figure 4
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
Cannabis
48h, 72h
8 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-2LIXDQ)
In Vitro Transporter Induction Experiment
Down Regulation was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system Jar
Results
figure 3
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM
Cannabis
72h
computerized image analysis system
9 . Down regulation of P-gp (mRNA) by Cannabidiol (id=NPDI-E7xthw)
In Vitro Transporter Induction Experiment
Down Regulation was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system Jar
Results
figure 5
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
mRNA expression
Cannabis
24h, 48h, 72h
qRT-PCR, 2 CT method
10 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-DZj9Zg)
In Vitro Transporter Induction Experiment
Down Regulation was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system BeWo
Results
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
10 µM and 15 µM
Cannabis
72h
computerized image analysis system
11 . Down regulation of P-gp transporter (protein) by Cannabidiol (id=NPDI-29ER6A)
In Vitro Transporter Induction Experiment
Down Regulation was detected. Cutoff used —
N/A
- P-gp (ABCB1)
Cell system BeWo
Results
figure 2
| Sample | Compound measured | Value | Measurement | Study sequence | Additional information | N replicates |
|---|
Experimental Conditions
Protein expression
Cannabis
48h, 72h
computerized image analysis system