Effect Of Broneolum Syntheticum Or Rhizoma Acori Talarinowii Drugs Of Causing Resuscitation By Administering Aromatic Drugs On Penetrating Blood-Brain Barrier Of The Naodesheng's Active Component

OBJECTIVESTo estabolish the methods for determination hydroxysafflor yellow A, puerarin, tetramethylpyrazine, ginsenosides Rg1 in rat plasma and brain; to inspect the effect of Broneolum syntheticum or Rhizoma acori talarinowii on the pharmacokinetics and the acrossing the blood-brain barrier of Naodesheng's main active constituent; to study the mechanism of drugs of causing resuscitation by administering aromatic drugs opening the blood-brain barrier.METHODS1. Dose and sample collection1.1 Male Sprague-Dawley rats (220-260g) were equally divided into three groups by randomized design according to their body weight: Naodesheng group (Ⅰ), Naodesheng compatibility Borneolum group (Ⅱ) and Naodesheng compatibility Rhizoma acori talarinowii group (III).1.2 According to the"Chinese Pharmacopeia"of 2005 edition, the daily dose of Borneolum and extrat of Rhizoma acori talarinowii was 28 mg·kg-1,3.0 g·kg-1, respectively. Each rat was intragastric administration Naodesheng which included HSYA 20 mg·kg-1, Puer 20 mg·kg-1, TMP 10 mg·kg-1, Rg120 mg·kg-1. The extract of Rhizoma acori talarinowii: Rhizoma acori talarinowii had been soaked for 30 min and destiled. Collection the mixed solution of volatile oil and water, adding tween 80 (content 1%). At last, metered volume to 3.0 g·mL-1 with water.1.3 Animals were fasted for 12h before experiments but had free access to water. Overnight-fasted rats were orally administered medicine suspended in 0.8%sodium carboxymethylcellulose solution. After oral administration of medicine, the rats were sacrificed by picking off eyeballs of rats at the time intervals of 10,20,30,45 min,1,1.5,2, 3,6,8,12 h, and the cerebra were collected and weighted. At each time point, six rats were sacrificed and sampled.2. Sample analysis2.1 Puerarin, tetramethylpyrazine, and ginsenosides Rg1 in rat plasma and brain were simultaneous determined by UPLC-MS/MS.2.2 The HSYA in rat plasma and brain was determined by HPLC.3. Data processingThe dates were analysised by SPSS 13.0 software. And then the of each group was tested by one-factor analysis of variance.RESULTS1 Simultaneous determination of puerarin, tetramethylpyrazine, and ginsenosides Rg1The interference of endogenous compounds were assessed by analyzing standard puerarin, tetramethylpyrazine and Rg1 with the retention times of 3.16,2.18,3.59,2.59 min for plasma and 3.14,2.17, 3.59,2.65 min for brain homogenate samples. Standard curves exhibited good linearity. The limits of quantitations in plasma and brain both were 10 ng-mL"1,12.5 ng-mL-1,10 ng-mL-1, respectively. The extraction in plasma and brain recovery both were more than 80.77%.The intra-and inter-day precisions of analysis were less than 10.27%in plasma, and the lower concentration were less than 16.12%, center and higher concentration were less than 11.88%in brain.2 Determination of HSYAUnder the optimized chromatographic conditions, HSYA and IS were separated well in rat plasma and brain homogenate with the retention times of 6.2 min and 11.9 min, respectively. Standard curves exhibited good linearity. The limits of quantitation in plasma and brain both were 40 ng·mL-1. The extraction in plasma and brain recovery were more than 85.07%,83.49%.The intra-day precisions of analysis were less than 11.2%,5.13% and the inter-day were less than 8.15%,9.14%, respectively.3 Animal experimentFor theⅠ,Ⅱ,Ⅲgroups, the AUC brain/AUC blood of HSYA were 0.69±0.044,0.98±0.054 and I.64±0.082; the AUC bmin/AUC blood of puerarin were 0.58±0.11,0.61±0.04 and 0.78±0.04; the AUC brain/AUC blood of tetramethylpyrazine were 0.0069±0.00085,0.0072±0.0010 and 0.014±0.0022. The AUC0→12 of Rg1 was 9289.98±1750.48, 13792.73±2065.88 and 15620.05±2936.68 in plasma, respectively, but could not be determinated in brain.CONCLUSIONS1 MethodsThe methods are rapid, sensitive, reproducible, accurate and suitable for analysis of the HSYA, Puer, TMP and Rg1 in rat plasma and brain to inspect the permeability of BBB.2 Animal experimentThe Borneolum enhance the HSYA, Puer, TMP penetrating BBB; Rhizoma acori talarinowii inhibit the absorption of HSYA and Puer, but enhance the absorptiong of TMP and Rg1. The Rhizoma acori talarinowii enhance the permeability of BBB, efficiently. The drugs of causing resuscitation by administering aromatic drugs can be messenger drug to aim the medicine to the brain.