Research have showed that lipophilic compounds Tanshinone I, Tanshinone IIA, Cryptotanshinone, and 15, 16dihydrotanshinone I had the ability to ameliorate memory decits induced by scopolamine, Tanshinone IIA and 2 Tanshinone IIB could bring about reduced amount of brain Letrozole infarct volume and the restoration of neurological function in an experimental model of stroke in mice, Cryptotanshinone could boost the cognitive skill in Alzheimers illness transgenic mice. Besides, Tanshinone I, Tanshinone IIA, and Cryptotanshinone were also located to become the substrates of P gp. However, it can be still unclear no matter whether Danshensu, a hydrophilic compound in Danshen, has got the prospective of crossing the BBB or could be the substrate of P gp. The current examine aims to research the part of P gp within the transport of Danshensu across the BBB by watching Danshensu concentration in plasma and brain tissue in rats. Danshensu was obtained from Shandong Luye Pharmaceutical Co., Ltd.. Verapamil was obtained from Shanghai Hefeng Pharmaceutical Co., Ltd.. Naproxen was obtained from National Institute to the Control of Pharmaceutical and Biological Products. Ethyl acetate was acquired Letrozole from Sinopharm Chemical Reagent Co., Ltd.. Acetonitrile was obtained from Merck. Forty-eight male Sprague Dawley rats weighing 220 20 g were provided by the Experimental Animal Center of Shandong Engineering Research Center for Natural Drugs, certicate number 20030020. All experimental procedures carried out in this study were performed in accordance with the rules for the Care and Use of Laboratory Animals of Yantai University. The subjects were kept with free access to food and water on a 12 h light/dark period. They certainly were housed in plastic cages and mapk inhibitor randomly divided into two groups with 24 animals in each group: the control group and the verapamil group. The subjects in the verapamil group were administered intraperitoneally with verapamil at a dose of 20 mg kg1. The subjects in the get a grip on group were treated with the same amount of normal saline. Ninety minutes later, all subjects were treated intravenously with Danshensu by tail vein. At 15 min, 30 min, and 60 min after Danshensu treatment, the animals were anesthetized with chloral hydrate and then 5 mL heparinized blood were collected from abdominal aorta and the rats were perfused with 100 mL of ice cold normal saline each. The brain was rapidly taken off the cranium and weighed. Then the brain was homogenized in 4 volumes of 0. 1 mol L1 ice phosphate buer. Three milliliters NSCLC of ethyl acetate was added into 200 uL of the homogenate. After vortexing for 3 min and centrifuging for 5 min, the supernatants were evaporated to dryness under a gentle nitrogen stream at 40 C. The deposits were resuspended in mobile phase. The blood samples were centrifugated for 10 min and plasma was separated. Plasma was treated as described for brain homogenate supernatants. The chromatographic separation was performed utilizing an Agilent 1100 Series HPLC system equipped with a vacuum degasser, a quaternary pump, an, and a column oven. The chromatographic separation was operate on a ODS C18 column. The mobile phase was acetonitrilewater. The pump was operated at a rate of 0. 2 mL min1. mapk inhibitor Separations were done at the temperature of 20 C. Mass spectrometric detection was performed utilizing a TSQ Quantum tandem mass spectrometer equipped with an electrospray ionization source. Quantication was performed using selected reaction monitoring of the transitions of m/z 197. 0?? m/z 135. 1 for Danshensu and m/z 229. 0?? m/z 170. 1 for the naproxen. The mass spectrum conditions were optimized as follows: spray voltage, 3000 V, sheath gas pressure, 30 psi, auxiliary gas pressure, 5 arbitrary device, capillary temperature, 350 C, collision induced dissociation voltage, 18 V, argon gas pressure, 1. 5 millitorr. Data acquisition was performed with Xcalibur computer software. Ionization was controlled in negative Selected Ion Monitoring mode. Sheath gas pressure was 30 kPa Letrozole and mapk inhibitor aux gas pressure was 5 kPa. Capillary temperature was 150 C. Ion sweep gas pressure was 0 kPa and Tube Lens oset was 105 eV. Data is expressed as means SEM. The statistical signicances of the data were determined using one of the ways analysis of variance followed closely by minimal Signicant Dierence testing. The P value. 05 was regarded as statistically signicant. Chromatogram of Danshensu. Figures 1 and 2 show the normal SRM chromatograms of the blank rat brain, brain spiked with Danshensu and naproxen, brain of Danshensu treated rat with spike of naproxen, blank rat plasma, plasma spiked with Danshensu and naproxen, plasma of Danshensu treated rat with spike of naproxen.
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