Goldenseal (Hydrastis canadensis) 43525850
Inhibition of P-Glycoprotein by HIV Protease Inhibitors Increases Intracellular Accumulation of Berberine in Murine and Human Macrophages CSV JSON

Background: HIV protease inhibitor (PI)-induced inflammatory response in macrophages is a major risk factor for cardiovascular diseases. We have previously reported that berberine (BBR), a traditional herbal medicine, prevents HIV PI- induced inflammatory response through inhibiting endoplasmic reticulum (ER) stress in macrophages. We also found that HIV PIs significantly increased the intracellular concentrations of BBR in macrophages. However, the underlying mechanisms of HIV PI-induced BBR accumulation are unknown. This study examined the role of P-glycoprotein (P-gp) in HIV PI-mediated accumulation of BBR in macrophages.

Methodology and Principal Findings: Cultured mouse RAW264.7 macrophages, human THP-1-derived macrophages, Wild type MDCK (MDCK/WT) and human P-gp transfected (MDCK/P-gp) cells were used in this study. The intracellular concentration of BBR was determined by HPLC. The activity of P-gp was assessed by measuring digoxin and rhodamine 123 (Rh123) efflux. The interaction between P-gp and BBR or HIV PIs was predicated by Glide docking using Schrodinger program. The results indicate that P-gp contributed to the efflux of BBR in macrophages. HIV PIs significantly increased BBR concentrations in macrophages; however, BBR did not alter cellular HIV PI concentrations. Although HIV PIs did not affect P- gp expression, P-gp transport activities were significantly inhibited in HIV PI-treated macrophages. Furthermore, the molecular docking study suggests that both HIV PIs and BBR fit the binding pocket of P-gp, and HIV PIs may compete with BBR to bind P-gp.

Conclusion and Significance: HIV PIs increase the concentration of BBR by modulating the transport activity of P-gp in macrophages. Understanding the cellular mechanisms of potential drug-drug interactions is critical prior to applying successful combinational therapy in the clinic.

PMID: 23372711

23372711

1 . Amprenavir P-gp Inhibition (id=NPDI-3z4F5w)

In Vitro Transporter Kinetics Experiment

Transport Activity was detected.  Cutoff used — not specified

berberine 19012197

  • P-gp (ABCB1)

Transfected / siRNA knock-out / injected cells MDCK transfected cells

Results

Figure 5b - reported value above is for 25 μM of Amprenavir vs control and is significant p< 0.05

Sample Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Amprenavir concentration tested: 5, 15, and 25  μM

Cell Culture Conditions

5

Assay Conditions

4 h

37 deg C

5 μM

2 . Lopinavir P-gp Inhibition (id=NPDI-VpSvoQ)

In Vitro Transporter Kinetics Experiment

Transport Activity was detected.  Cutoff used — Not specified

berberine 19012197

  • P-gp (ABCB1)

Transfected / siRNA knock-out / injected cells MDCK transfected cells

Results

Figure 5b - reported value above is for 25 μM of lopinavir vs control and is significant p< 0.05

Sample Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Concentration of HIV PI's were 5, 15, and 25 μM

Cell Culture Conditions

5

Assay Conditions

4 h

37 deg C

5 μM

3 . Ritonavir P-gp Inhibition (id=NPDI-a3Bf6g)

In Vitro Transporter Kinetics Experiment

Transport Activity was detected.  Cutoff used — not specified

berberine 19012197

  • P-gp (ABCB1)

Transfected / siRNA knock-out / injected cells MDCK transfected cells

Results

Figure 5 - reported value above is for 25 μM of ritonivir vs control and is significant p< 0.05

Sample Compound measured Measurement Value Study sequence Additional information N replicates

Experimental Conditions

Concentration of HIV PI's were 5, 15, and 25 μM

Cell Culture Conditions

5

Assay Conditions

4 h

37 deg C

5 μM