rn210451  (OriGene)

Journal: Molecular metabolism

Article Title: Bile acids are important direct and indirect regulators of the secretion of appetite- and metabolism-regulating hormones from the gut and pancreas.

doi: 10.1016/j.molmet.2018.03.007

Figure Lengend Snippet: Figure 4: Bile acids stimulate GLP-1, NT, and PYY secretion by activation of basolateral, but not luminal, TGR5 receptors. Data are shown as means 1 SEM. All X-Y plot data with exception of C, D, F, I and J are from isolated perfused rat small intestine. A1,2: Effects of luminal and vascular administration of a poorly absorbable TGR5 agonist on GLP-1, NT and PYY secretion, n¼6. B1,2: Effects of luminal and vascular administration of GW4064 (a FXR agonist) on GLP-1, NT and PYY secretion, n¼6. C,D: Activation of TGR5 in response to different bile-acids in cells transfected with either the human TGR5 receptor (C) or rat TGR5 receptor (D). E: Co-localization of TGR5 and GLP-1/PYY in rat small intestine. F: Expression of TGR5 in isolated pancreatic a-, b- or d-cells or intestinal L-cells from the mouse, n¼3. G: Plasma GLP-1 (total) concentrations in response to a complex BA-mix (n¼6), match total conc. of UDCA (n¼8) or 0.9%NaCl (neg. control, n ¼ 3). H1,2: Effects of vascular TUDCA, TCDCA or TDCA on GLP-1, NT and PYY secretion, n¼6. I and J: Effects of a complex BA-mix on the secretion of GLP-1 and PYY from isolated perfused mouse small intestine. I: WT mice, J: TGR5 KO mice, n ¼ 6. In all perfusion experiments, bombesin (BBS) was administered in the end of the experiment and used as positive control. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: 35,000 COS-7 cells per well were seeded in 96-well plates coated with poly-Dlysine and modified for either human or rat TGR5 expression using a transient calcium phosphate precipitation transfection procedure [21], using a pCMV6-XL5 or pCMV6-Entry vector, respectively (Cat. no. SC123312 and RN210451, OriGene, Technologies Inc., Rockville, MD).

Techniques: Activation Assay, Isolation, Transfection, Expressing, Clinical Proteomics, Control, Positive Control

Journal: Molecular metabolism

Article Title: Bile acids are important direct and indirect regulators of the secretion of appetite- and metabolism-regulating hormones from the gut and pancreas.

doi: 10.1016/j.molmet.2018.03.007

Figure Lengend Snippet: Figure 5: TGR5 activation or bile acids has no direct effects on glucagon or insulin secretion. Data are shown as means ? 1 SEM. All data are from isolated perfused rat pancreas. A. Glucagon (1,2) and insulin (3,4) secretion at low glucose (3.5 mmol/L) in response to the TGR5 agonist RO9272296 or a complex BA-mix. B: Glucagon (1,2) and insulin (3,4) secretion at high glucose (10 mmol/L) in response to the TGR5 agonist RO9272296 or a complex BA-mix. L-arginine (Arg) was included at the end of all experiment and used a positive control. n ¼ 6 for all experiments. *P < 0.05, **/##P < 0.01, ***/###P < 0.001, ****P < 0.0001. Stars indicate significance compared to baseline and hashes indicate significance between treatments.

Article Snippet: 35,000 COS-7 cells per well were seeded in 96-well plates coated with poly-Dlysine and modified for either human or rat TGR5 expression using a transient calcium phosphate precipitation transfection procedure [21], using a pCMV6-XL5 or pCMV6-Entry vector, respectively (Cat. no. SC123312 and RN210451, OriGene, Technologies Inc., Rockville, MD).

Techniques: Activation Assay, Isolation, Positive Control

Journal: Molecular metabolism

Article Title: Bile acids are important direct and indirect regulators of the secretion of appetite- and metabolism-regulating hormones from the gut and pancreas.

doi: 10.1016/j.molmet.2018.03.007

Figure Lengend Snippet: Figure 6: Proposed model of bile acid stimulated secretion of appetite and metabolism regulating hormones. (1) Food intake, and in particular fat consumption, stimulates the secretion of bile acids into the upper small intestine by CCK-mediated contraction of the gallbladder. (2) In addition to their well-known role in facilitating fat absorption (by micelle formation) bile acids activate TGR5 receptors which are located at the basolateral membranes of the enterocytes and therefore are activated secondary to bile acid (BA) absorption. Upon activation, the secretion of GIP, GLP-1, NT, and PYY is stimulated. Conjugated bile acids are absorbed through the secondary active transporter ileal-bile acid transporter (IBAT) which is predominantly expressed in the lower part of the small intestine, whereas unconjugated bile acids (which are more lipophilic) spontaneously cross the intestinal mucosal layer. (3) Eliminating bile acid absorption by BA-sequestrants (which cross bind both conjugated and unconjugated BAs into large unabsorbable complexes) or by direct IBAT inhibition (which attenuates the absorption of conjugated BAs) therefore eliminates BA-stimulated gut hormone secretion. (4) Collectively, the absorption mechanisms results in a very efficient BA absorption so about 95% of the secreted bile acids are returned to the liver through the enterohepatic circulation. (5) The majority of the returned bile acids are extracted by the liver (where they are reconjugated and rehydroxylated, allowing the same pool of bile acids to be secreted several times during the day), (6) Only 3e10% pass the liver and eventually ends up in the systemic circulation, (7) The pool is further diluted with a factor of about three since the hepatic return constitutes about 1/3 of the total venous return. (8) Therefore, only a small fraction of the secreted bile acid makes it to the systemic circulation.

Article Snippet: 35,000 COS-7 cells per well were seeded in 96-well plates coated with poly-Dlysine and modified for either human or rat TGR5 expression using a transient calcium phosphate precipitation transfection procedure [21], using a pCMV6-XL5 or pCMV6-Entry vector, respectively (Cat. no. SC123312 and RN210451, OriGene, Technologies Inc., Rockville, MD).

Techniques: Activation Assay, Inhibition