Journal: Biochemical pharmacology

Article Title: Rifampicin and its derivatives: stability, disposition, and affinity towards pregnane X receptor employing 2D and 3D primary human hepatocytes.

doi: 10.1016/j.bcp.2024.116500

Figure Lengend Snippet: Fig. 6. The effect of forced OATP1B1 expression on induction of CYP3A4 gene reporter by rifampicin and its derivatives. HepG2 cells were transfected with p3A4-luc reporter construct, pSG5-hPXR, and pRL-TK along with pOATP1B1 or pcDNA3 for 24 h and then treated with rifampicin and its derivatives at indicated concen trations for 12 h. Data are presented as the fold change in Renilla luciferase-normalized firefly luciferase activities to corresponding DMSO control (set as 1). The differences in induced p3A4-luc reporter construct activities between HepG2 cells transfected with pOATP1B1 and those transfected with empty pcDNA3 plasmid were analyzed by an unpaired t-test. * indicates p < 0.05 and ** indicates p < 0.01.

Article Snippet: Expression vectors for human OATP1B1 transporter (200 ng/well, SC116071, NM_006446) and human MDR1 transporter (200 ng/well, OHu102681, NM_001348945.1) were acquired from OriGene (Rockville, Maryland, USA) and Genscript (Piscataway, New Jersey, USA), respectively.

Techniques: Expressing, Transfection, Construct, Luciferase, Control, Plasmid Preparation

Journal: Biochemical pharmacology

Article Title: Rifampicin and its derivatives: stability, disposition, and affinity towards pregnane X receptor employing 2D and 3D primary human hepatocytes.

doi: 10.1016/j.bcp.2024.116500

Figure Lengend Snippet: Fig. 8. AADAC, OATP1B1, OATP1B3, and MDR1 mRNA expression changes in 3D/2D PHHs following treatment with rifampicin. 3D PHH spheroids (donors 1–3) were treated with rifampicin (10 µM) for an indicated time ranging from 4 to 168 h. In the case of 2D PHHs, donors 1 and 2 were only treated with rifampicin (10 µM) for 48 h and 24 h, respectively. AADAC (A), OATP1B1 (B), OATP1B3 (C), and MDR1 (D) mRNA expression levels were analyzed by RT-qPCR. The data are shown as the fold change expression relative to the corresponding DMSO control at the same time (set as 1). ANOVA with Dunnett’s post hoc test was used to compare time- dependent rifampicin-induced changes in mRNA expression relative to time 0 (set as 1) in 3D PHHs. * indicates p < 0.05, ** indicates p < 0.01, and *** indicates p < 0.001.

Article Snippet: Expression vectors for human OATP1B1 transporter (200 ng/well, SC116071, NM_006446) and human MDR1 transporter (200 ng/well, OHu102681, NM_001348945.1) were acquired from OriGene (Rockville, Maryland, USA) and Genscript (Piscataway, New Jersey, USA), respectively.

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: The Journal of Physiology

Article Title: Glutamate cycling may drive organic anion transport on the basal membrane of human placental syncytiotrophoblast

doi: 10.1113/jp270743

Figure Lengend Snippet: Figure 2. Expression of OATs and OATPs in term human placenta and primary cytotrophoblast A, rtPCR amplification of cDNA from term human placenta, control tissue (kidney or liver) and no template control (NTC) for the OATs. B, rtPCR amplification of cDNA from term human placenta, control tissue (kidney, brain or liver) and NTC for OATP1A2, OATP1B3, OATP3A1, OATP4C1. C, rtPCR amplification of cDNA from term human placenta and NTC for OATP2A1, OATP4A1 and OATP2B1. D, rtPCR amplification of cDNA from term human placenta, control tissue (kidney) and NTC for AGT-1. E, rtPCR amplification of cDNA from primary term human cytotrophoblast culture and in NTC for OATP2B1, OATP2B1, OATP4A1 and OAT4. plac, placenta; kid, kidney; liv, liver; cyto, cytotrophoblast.

Article Snippet: Where no expression was detected in placenta, primers were tested using cDNA from other cell lines or tissues thought to express the gene of interest as outlined in Table 1. cRNA synthesis for microinjection into oocytes Plasmids containing the cDNA of human OAT4 or human OATP2B1 obtained from OriGene (OriGene Technologies Inc., Rockville, MD, USA) were linearised using restriction enzymes (Promega, Southampton, UK).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Control

Journal: Biochimica et biophysica acta. Molecular basis of disease

Article Title: Animal Models to Study Bile Acid Metabolism

doi: 10.1016/j.bbadis.2018.05.011

Figure Lengend Snippet: Genetic Mouse Models Used to Study Bile Acid Biosynthesis and Metabolism

Article Snippet: Slco1a/1b KO (Oatp1a/1b) Humanized SLCO1B1 Slco1b2 KO (Oatp1b2) , Constitutive Slco1a/1b gene cluster KO [(FVB.129P2-Del(Slco1b2-Slco1a5)1Ahs]; Taconic model #10707; ↑serum unconjugated BAs; ↑serum conjugated bilirubin [ 152 , 180 ] Tg mouse expressing human OATP1B1 driven by apoE hepatic control region crossed into Slco1a/1b KO (FVB/129; Taconic model #10708); ↔serum conjugated bilirubin [ 152 ] Constitutive Slco1b2 KO (backcrossed into C57Bl/6J); ↑serum unconjugated BAs; ↓hepatic CA clearance [ 182 ].

Techniques: Expressing, Control, Mutagenesis, Knock-In, Activity Assay

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Generation of humanized OATP1B1 rats by CRISPR/Cas9. (A) The strategy for the generation of the h OATP1B1 rat model. The donor template (2361 bp) was inserted at the second exon. Mutations in the F0 generation (B) and F2 generation (C). H 2 O, negative control; WT, blank control; Arrow, mutant band. The expression of Slco1b2 (D) and SLCO1B1 (E) in WT and h OATP1B1 rats. Data are shown as mean ± SD ( n = 6). N.D., not detected. ∗∗∗ P < 0.001 compared with the WT group. (F) The protein expression of OATP1B1 in the liver (Western blot). (G) OATP1B1 is highly expressed on the sinusoidal membrane of the h OATP1B1 rat liver (shown in brown).

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques: CRISPR, Negative Control, Control, Mutagenesis, Expressing, Western Blot, Membrane

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Physiological characterization of h OATP1B1 rats. (A)–(D) Blood lipids indexes in the serum of WT and h OATP1B1 rats, including TG, TC, HDL-C, and LDL-C. (E) H&E staining of liver tissue sections. Scale bars, 50 μm in length. (F)–(K) Indicators of liver function in the serum of WT and h OATP1B1 rats, including TP, ALB, AST, ALT, AST/ALT, and ALP. (L)–(N) The contents of TBIL, DBIL, and TBA in the serum of WT, h OATP1B1, and OATP1B2 KO rats, respectively. Values are shown as mean ± SD ( n = 6). ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques: Staining

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Effects of SLCO1B1 gene insertion on bile acid-related pathway. (A) Differential abundance score and size of perturbed metabolic pathways between WT and h OATP1B1 rats. Size means the number of differential metabolites included in the pathway (KEGG database matched results). (B) Heat plot of the differential metabolites (bile acid pathway) between WT and h OATP1B1 rats ( n = 4).

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Pharmacokinetic profiles of statins in WT, h OATP1B1, and OATP1B2 KO rats. (A) Pitavastatin, 5 mg/kg. (B) Rosuvastatin, 10 mg/kg. (C) Fluvastatin, 5 mg/kg. All statins were administered intragastrically and the concentration of statins was quantified by LC–MS/MS. The concentration corresponding to each time point on the curve was presented as mean ± SD ( n = 6).

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Pharmacokinetic parameters of pitavastatin, rosuvastatin, and fluvastatin in WT, h OATP1B1, and OATP1B2 KO rats.

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques:

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: Transport of statins in OATP1B1 overexpression cells. (A) Green fluorescence after transfection of HEK293T cells for 24 h. Scale bars, 2000 μm. (B) Protein expression of OATP1B1 (Western blot). The concentration of pitavastatin (C), rosuvastatin (D), and fluvastatin (E) in cell lysis. Data are shown as mean ± SD ( n = 4). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques: Over Expression, Fluorescence, Transfection, Expressing, Western Blot, Concentration Assay, Lysis

Journal: Acta Pharmaceutica Sinica. B

Article Title: Construction and characterization of a humanized SLCO1B1 rat model with its application in evaluating the uptake of different statins

doi: 10.1016/j.apsb.2023.12.019

Figure Lengend Snippet: The binding modes and enzyme–substrate interactions of pitavastatin, rosuvastatin, and fluvastatin with OATP1B1 in the cavity of extracellular domains (A) and transmembrane helices (B).

Article Snippet: The OATP1B1 rabbit primary antibody (A15783) was bought from ABclonal Technology (Wuhan, China), and the anti-GAPDH antibody (ab8245) was purchased from Abcam (Cambridge, UK).

Techniques: Binding Assay