Drawer9parade: Створена сторінка: 2012b). Briefly, aortic strips (two preweighed whole sections of aortae for each single measurement) were initially incubated with PSS at 37��C for 30 min,...

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Створена сторінка: 2012b). Briefly, aortic strips (two preweighed whole sections of aortae for each single measurement) were initially incubated with PSS at 37��C for 30 min,...

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2012b). Briefly, aortic strips (two preweighed whole sections of aortae for each single measurement) were initially incubated with PSS at 37��C for 30 min, followed by a 10 min exposure to PSS, AA (30 ��m) or PGH2 (10 ��m) in 1000 ��l PSS (37��C). Thereafter, the reaction solution was pipetted into another tube, and the proportion of metabolites in tissues was further extracted with acetone, followed by drying with a stream of N2 and dissolving with the original reaction solution. Sample or standard solution (25 ng 6-keto-PGF1��, or 6-keto-PGF1��, TxB2, PGF2��, PGE2 and PGD2 at the indicated amounts; Cayman Chemical) further underwent solid-phase extraction as described elsewhere (Liu et al. 2012b). The HPLC system (Agilent 1100; Agilent [http://www.selleckchem.com/products/GDC-0449.html www.selleckchem.com/products/GDC-0449.html] Technologies, Santa Clara, CA, USA) was equipped with an HPLC column (Symmetry? C18, 3.5 ��m, 2.1 mm �� 100 mm; Waters, Milford, MA, USA). The mobile phase was methanol/10 mm ammonium acetate (50/50, pH 7.5) with a flow rate of 0.2 ml min?1. The amount of 6-keto-PGF1�� was calculated from the area of signal relative to that of a standard (25 ng), and was expressed in nanograms per milligram of wet tissues. The expression of COX-1, COX-2 or ��-actin in abdominal aortae was detected by real-time PCR. Preparation of RNA was performed using an RNAiso? Plus kit (TaKaRa, Dalian, China), [https://en.wikipedia.org/wiki/Moroxydine Moroxydine] according to the manufacturer's instructions. First strand cDNA was synthesized using total RNA (250 ng) and Oligo(dT)15 primers (TakaRa). The PCR primers for COX-1 were 5��-CCA ACT GTA CCA TCC CTG AG-3�� (sense) and 5��-AAT TCC CAG AGC CAG TAT CC-3�� (antisense); those for COX-2 and ��-actin were described previously (Liu et al. 2012b, 2013). Real-time PCR was performed using the SYBR? PrimScript? RT-PCR kit (TaKaRa). Data were expressed as means �� SEM from n numbers or pools of vessels from different animals. Student's unpaired t test (two tailed) was used to compare the difference between two means for statistical evaluation. When more [http://www.selleckchem.com/products/Methazolastone.html Temozolomide ic50] than two means were compared, a one-way ANOVA followed by Dunnett's post hoc test or two-way ANOVA with Bonferroni's post hoc test was used. A value of P

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Drawer9parade: Створена сторінка: 2012b). Briefly, aortic strips (two preweighed whole sections of aortae for each single measurement) were initially incubated with PSS at 37��C for 30 min,...