Fluoxetine hydrochloride (FLX) was purchased from Sigma-Aldrich (St Louis, MO) and dissolved in DMSO and diluted in 0.9% sterile saline (NaCl) with a final concentration of 1% DMSO. Ketamine hydrochloride was purchased from the Hengrui Pharmaceutical Co., Ltd. (Jiangsu, China) and dissolved in 0.9% sterile saline (NaCl). The dose of ketamine or FLX at 10 mg/kg was chose as described before and was administered intraperitoneally (i.p.) (Garcia et al., 2008 (link); Fernandez Macedo et al., 2013 (link)). Control animals received same amount of 0.9% sterile saline i.p. or vehicle i.p. (DMSO 1%).
Fluoxetine hydrochloride flx
Manufactured by Merck Group
Sourced in United States
Fluoxetine hydrochloride (FLX) is a white or almost white crystalline powder. It is soluble in water, freely soluble in alcohol, and slightly soluble in acetone and in dichloromethane.
Automatically generated - may contain errors
Lab products found in correlation
Sucrose, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl, by Merck Group (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
Glacial acetic acid, by Avantor (1 mentions)
Sodium hydroxide (naoh), by Thermo Fisher Scientific (1 mentions)
Sodium acetate, by Thermo Fisher Scientific (1 mentions)
Milli q system, by Merck Group (1 mentions)
Boric acid, by Merck Group (1 mentions)
9 protocols using fluoxetine hydrochloride flx
1
Ketamine and Fluoxetine Administration Protocol
2
Synthesis and Administration of pBBG and MeGFN
3
Extraction and Purification of Apocynum venetum Leaf Extract
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Apocynum venetum leaf extract was obtained from dried leaves as previously described [27 (link)]. Briefly, Apocynum venetum leaves (100 g) were refluxed for 1 h in aqueous ethanol (70%, v/v, 60 mL) twice, and the combined alcoholic extract evaporated to dryness (28 g). The extract (13.5 g) was dissolved in hot water (200 mL) and the pH adjusted to 3.0 with sulfuric acid and then filtered. The filtrate underwent chromatography on a Diaion HP-20 (3.6 cm i.d. × 18 cm; Sigma-Aldrich, St. Louis, MO, USA) column and eluted with water (200 mL) followed by aqueous ethanol (70%, v/v, 200 mL). The aqueous ethanol fraction was collected and evaporated to dryness to obtain AVLE (4.2 g). Fluoxetine hydrochloride (FLX) was obtained from Sigma (St. Louis, MO, USA). Sucrose was purchased from Sigma-Aldrich (St. Louis, MO, USA).
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Apocynum venetum leaf extract was obtained from dried leaves as previously described [27 (link)]. Briefly, Apocynum venetum leaves (100 g) were refluxed for 1 h in aqueous ethanol (70%, v/v, 60 mL) twice, and the combined alcoholic extract evaporated to dryness (28 g). The extract (13.5 g) was dissolved in hot water (200 mL) and the pH adjusted to 3.0 with sulfuric acid and then filtered. The filtrate underwent chromatography on a Diaion HP-20 (3.6 cm i.d. × 18 cm; Sigma-Aldrich, St. Louis, MO, USA) column and eluted with water (200 mL) followed by aqueous ethanol (70%, v/v, 200 mL). The aqueous ethanol fraction was collected and evaporated to dryness to obtain AVLE (4.2 g). Fluoxetine hydrochloride (FLX) was obtained from Sigma (St. Louis, MO, USA). Sucrose was purchased from Sigma-Aldrich (St. Louis, MO, USA).
4
Synthesis and Administration of pBBG and MeGFN
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We synthesized pBBG (see McMurray et al. 2015)15 and MeGFN (see supplemental materials ) based on previously described methods (see Thornalley et al. 1996; Kawatani et al. 2008; Kanoh et al. 2013)5 (link),16 (link),17 (link). For the TST and acute FST mice received pBBG (50 mg/kg in 8% DMSO/18% Tween80 in H2O), MeGFN (12.5, 25 or 50 mg/kg in 4%DMSO/9%Tween80 in H2O) or their corresponding vehicle by I.P. injection 2 hours before testing. For the cFST, CMS and OBX, minipumps were filled with pBBG, MeGFN, or vehicle (50% DMSO, 50% PEG400) and inserted into a small subcutaneous incision made on the back18 (link). Fluoxetine hydrochloride (FLX; Sigma-Aldrich, St. Louis, MO) was delivered via the drinking water in opaque water bottles at a concentration of 160mg/L to achieve a dose of 18 mg/kg/day19 .
5
Fluoxetine's Effects on Behavioral Assays
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Fluoxetine hydrochloride (FLX, a selective serotonin reuptake inhibitor dissolved in 0.9% saline; 5 mg/kg i.p.; Sigma Aldrich, MO) was injected intraperitoneally 30 min before the 2AFC-VDT, open field test and water intake measurement. The dose of the drug was determined on the basis of a previous study [29 (link)].
6
Exploring Neuroprotective Mechanisms with Xiaoyao Pills
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Xiaoyao Pills (TaiJi, China), LPS (Escherichia coli 055:B5), fluoxetine hydrochloride (FLX), and DAPI were provided by Sigma (St. Louis, MO). Rabbit anti-NeuN antibody (1 : 50; Cat. No. #24307S), mouse anti-BrdU antibody (1 : 1400; Cat. No. #5292S), rabbit anti-TrkB antibody (1 : 1000; Cat. No. #4603), rabbit anti-CREB antibody (1 : 1000; Cat. No. #9197S), rabbit anti-p-CREB antibody (1 : 1000; Cat. No. #9198S), and rabbit anti-β-tubulin antibody (1 : 1000; Cat. No. #2128) were all from Cell Signaling Technology. Rabbit anti-synaptophysin polyclonal antibody (1 : 1000; Cat. No. #17785-1-AP) was from Proteintech. Rabbit anti-GAPDH antibody (1 : 1000; Cat. No. #GB11002) was from Servicebio. Goat anti-mouse IgG-FITC (1 : 100; Cat. No. #10) and goat anti-rabbit IgG/Cy3 (1 : 100; Cat. No. #AG04017512) were both from Absin.
7
Antioxidant and Pharmaceutical Evaluation
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1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), ascorbic acid (AA), and fluoxetine hydrochloride (FLX) were obtained from Sigma-Aldrich (USA). All other chemicals used are of analytical grade were purchased commercially from local vendors and were utilized without further purification.
8
Determination of Fluoxetine and Metabolite
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Fluoxetine hydrochloride (FLX; lot #SLBL4347V) and its primary active metabolite, norfluoxetine hydrochloride (NFLX), were purchased from Sigma-Aldrich. Sodium acetate buffer (0.050 M) was prepared from Sodium acetate (Fisher Scientific, Inc.) and glacial acetic acid (VWR brand). Borate buffer (0.1 M) was prepared from boric acid, H3BO4 (Sigma) and sodium hydroxide (Fisher Scientific). Solvents were HPLC-grade acetonitrile (Pierce) and water purified using a Milli-Q system (Millipore Corporation). Stir bar sorptive extraction (SBSE) was performed using GERSTEL-Twister sorptive stir bars (GERSTEL Gmbh & Co. KG) obtained from Agilent Technologies. The stir bars are 10-mm long and are coated with a 0.5-mm film thickness of polydimethylsiloxane (PDMS). Extractions were conducted in Fisherbrand 21 × 70-mm amber glass vials. Desorptions were performed in Varian 4.0-ml clear glass vials with PTFE/sil septa containing Agilent 400-µl glass inserts.
9
Fluoxetine Exposure in Pregnant Mice
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Between E15 and P12, pregnant dams were administered fluoxetine hydrochloride (Flx; Sigma-Aldrich, Saint Louis, MO, USA) in their drinking water at a dose of 25mg/kg/day as previously described [28] . The Flx concentration in the water was based on the mouse's weight at the time and the mouse's water consumption for the preceding 48 hours. This concentration was re-calculated every 48h to control for weight gain during the gestation period. This treatment protocol has been demonstrated to lead to pup brain levels of Flx and its metabolite norfluoxetine that fall within the range observed in post-mortem brain tissue of humans who took this medication [28] .
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