Phosphocreatine ( PCr), a natural high energy phosphate, plays a pivotal role in maintaining energy homeostasis of the body. Exogenous PCr has been developed as a cardio-protective drug and extensively used in treatment of cardiovascular diseases.PCr has special chemical structure,which confers much bioinformation on it,and becomes a multitarget-directed drug.Since the 21st century,with rapid development of molecular biology,the multiple target action mechanisms of PCr have continually gained elucidation,including energy-related and non-energy-related mechanisms,intracellular and extracellular mechanisms,which are leading to its extensive clinical applications in cardiovascular diseases.Based on author' s research and published literatures,this article reviews the research progress in multiple target action mechanisms of PCr,including energy supply,membrane stabilization, anti-platelet aggregation, electrophysiology, enzyme inhibition and protection of mitochondria, antiapoptotic effect,etc.

Objective To investigate effects of diabetes mellitus (DM) on pharmacokinetics (PK) of cephradine (CED).Methods DM model was induced by iv.alloxan 60 mg·kg-1.A reversed phase HPLC internal standard method was developed for measurement of CED plasma concentration.After blood collection,rats were sacrificed to collect kidneys for calculating kidney index(KW/BW).DM and normal control (CTL) rats were randomly assigned to receive iv.or ig.CED at a dose of 180 or 90 mg·kg-1.The 3p97 program was used to calculate PK parameters.Results The developed HPLC method was validated to have high specificity,precision,recovery and good storage stability,and met requirements for PK study of CED.The CED in rats of both DM and CTL groups showed the iv.two-compartment PK and ig.one-compartment PK and followed the first-order kinetics.Following iv.dosing,a remarkably decreased t1/2β and MRT,increased CLt were evident in DM group as compared with CTL group (P ＜ 0.05).After ig dosing,a significant decrease in t1/2k and t a remarkable increase in CLt and Cm=were observed for DM group as compared with CTL group (P ＜ 0.05).The DM rats showed a trend of decreased t1/2ka vs CTL rats.There was no significant difference in the oral bioavailability between the two groups (P ＞ 0.05).KW and KW/BW in DM group were increased remarkably compared with CTL group (P ＜ 0.05).Conclusion The DM vs CTL results in faster absorption and elimination of CED in rats,but does not have significantly affect in oral bioavailability.The compensatory hypertrophy and hyperfunction of early-stage diabetic kidneys may constitute one of causes of quick elimination of CED in rats with DM.

With the deepening of modernization of traditional Chinese medicine (TCM) and continuing emergence of new theories, methods and techniques, a very rapid and significant development has been achieved in the pharmacokinetics (PK) of TCM. This paper reviews the main research progresses of PK of TCM, including integrated PK of multiple effective components of TCM, fingerprint PK of TCM, novel dosage form PK of TCM, polysaccharide PK of TCM and drug interactions of TCM; and further sets up the prospects.

ObjectiveTo observe the relationship between neuronal function score and pathological changes of fluid percussion brain injury in rats and to explore their clinical significances.MethodsThe fluid percussion models of brain injury in rats were established by using the improved device with three kinds impact pressure such as 0.1 MPa,0.2 MPa,0.3 MPa,and vital signs and mortality rate were observed.Behavior changes,brain water content,histological changes were observed by Shapira and Wahld method,dry-wet measure,light microscopy at 1 h,6 h,12 h,24 h,3 d and 7 d after operation respectively.ResultsThe animals accepted impact pressure of 0.1 MPa showed temporary hypopnea with mortality rate of 2.08％,those of 0.2 MPa suffered apnoea of ( 10.88 ±2.69 ) s with mortality rate of 4.17％ and those of 0.3 MPa suffered apnoea of ( 20.60 ± 3.02 ) s with mortality rate of 16.67％.As the impact pressure increased,nervous function score minimumly decreased to (7.17 ±0.75) of 0.1 MPa group,(4.83 ± 0.75 ) of 0.2 MPa group and (2.67 ± 0.52) of 0.3 MPa group respectively,and recovered more slowly.Brain water content maximumly reached to (81.12 ± 0.03 )％,(82.74 ± 1.11 )％ and (83.89± 0.04) ％ at time point of 24 h respectively.The brain injury was involved in the outer layer of cerebral cortex,hippocampal formation and brain stem respectively and histological observation verified above findings.Conclusion Light,moderate and heavy fluid percussion brain injury in rats have more and more low nervous function scores,which have positive relationship with more and more serious pathological changes.

LC-MS/MS method was used to simultaneously determine anti-oxidative active catechins EGCG, ECG, EGC and EC in plasma of rats treated with tea polyphenols (TP). The integrated plasma concentration (C') of TP was calculated by means of self-defined weighing coefficient based on percent AUC of individual components, thereby assessing integrated pharmacokinetic (PK) parameters of TP via log C'-T curve. The anti-free radical effects of TP were estimated using inhibitory rate of drug-containing serum collected at different times from rats against in vitro lipid peroxidation of mouse liver homogenate. The obtained E-T curves were used to calculate anti-free radical pharmacodynamic (PD) parameters of TP. E-logC and E-log C' plots and linear regression were carried out in order to obtain the correlation coefficient (R2). The results indicated that the log C'-T curves of TP, which could be best described by three-compartment model, corresponded to elimination rule of iv administration of drugs. The integrated PK parameters showed that TP was distributed in body rapidly and widely, and eliminated from deep compartment slowly. From comparison of R2 values and consistence of C'-T course and E-T course, it was evident that TP integrated PK behaviors correlated much better with its PD behaviors than individual active components, and thus demonstrated that integrated PK parameters could characterize to maximal extent holistic disposition of Chinese herbal drugs and reflect residence properties of holistic effective substances in biological body.

ObjectiveTo observe the relationship between neuronal function score,brain edema and aquaporin4(AQP4) expression of fluid percussion brain injury in rats.MethodsThe fluid percussion models of brain injury of rats were established by using the improved device.Nervous function score,brain water content,histological changes,AQP4 expression were observed by Shapira and Wahld method,dry-wet measure,light microscopy,immunohistochemistry and western blot at 1 h,6 h,12 h,24 h,3 d and 7 d after operation respectively.ResultsNervous function score in TBI group decreased at 12 h( 11.17 ± 1.32),reached its minimum at 24 h( 10.17± 0.75),and recoved partially at 3rd day( 10.66 ± 1.37 ).The water content obviusly increased in those of TBI group at 12h( (80.27 ±1.47)％ ),reached its peak at 24h( (82.19 ±0.97)％ ),and then began to drop at 3d ( (8 1.74 ± 1.69 ) ％ ),while Western blot showed that AQP4 immunoreactive expression obviusly increased at 12 h (OD:0.65 ±0.05),reached its maximum at 24h( OD:0.72 ±0.08),and decreased at 3d( OD:0.56 ±0.07),and immunohistochemistry showed the same trendency of AQP4 expression as that of Western blot.The linear regression analysis indicated that nervous function score had a negtive correlation with expression of AQP4 in edematous brain and change of brain water content respectively ( r =- 0.615,P ＜ 0.01 ; r =- 0.605,P ＜ 0.05 ).ConclusionNervous function score of fluid percussion brain injury in rats decrease,has negative relationship with brain edema and AQP4 expression,and possible mechanisms is that AQP4 is indirectly involed in nerve function impairment by mediating brain edema.

This article is report the study of the pharmacokinetics and metabolic disposition of exogenous phosphocreatine (PCr) in rats by means of an ion-pair HPLC-UV assay. PCr and its metabolite creatine (Cr) and related-ATP in rat plasma and red blood cell (RBC) were simultaneously determined. A blank plasma and RBC were initially run for baseline subtraction. Plasma and RBC samples were deproteinized with 6% PCA prior to HPLC. Following i.v. administration of PCr 500 mg x kg(-1) and 1 000 mg x kg(-1) the C-T curve could be described by the two-compartment model with t1/2beta 22.5-23.3 min, V(d) 0.956 4-0.978 6 L x kg(-1), CL 0.029 L. kg(-1) x min(-1). The Cr as PCr degraded product appeared as early as 2 min post i.v. dosing with t(max) 20 min, t1/2kappa (m) 40.6-42.7 min and f(m) 60%-76%. After po administration of PCr, the parent drug in plasma was undetectable, but the metabolite Cr was detected with t(max) 65-95 min, t1/2kappa (m) 56.0-57.7 min, metabolite-based bioavailability F(m) 55.02%-62.31%. PCr i.v. administration resulted in significant elevation of ATP level in RBC but not in plasma, the related-ATP in RBC was characterized by t(max) 68-83 min, t1/2kappa 49-52 min. In RBC no exogenous PCr was found but Cr was detected following i.v. administration of PCr, with the t(max) 120 min and t1/2k (m) 70 min for Cr. The above results indicate that PCr eliminates and bio-transforms in body very rapidly; K > K(m) confers ERL, instead of FRL, type upon the metabolic disposition of Cr. Following po administration of PCr, the degraded product Cr is absorbed but not the parent drug PCr. The formed Cr can be accounted for by most of i.v. and po PCr. Intravenous dosing leads apparently increased and sustained Cr and related-ATP concentration in RBC.

Objective To develop a liquid chromatography technique coupled with tandem mass spectrometry(LC-MS/MS)for simultaneous determination of four active catechins EGCC,ECG,EGC,and EC of tea polyphenols(TP)in rat plasma in order to further study its multi-component pharmacokinetics.Methods Following a single step liquid-liquid extraction of plasma samples with ethyl acetate,the four catechins were separated on a Hypersil ODS C18 column using an isocratic mobile phase composed of methanol-water(30:70).The detection using a mass spectrometer was performed under negative ESI in the MRM mode.The analytes were identified by reference to both MRM and tR values and quantified using peak area internal standard method.Results The method was shown to be specific without interference from matrix,metabolites,and impurities present in TP raw material and to be sensitive with LOD and LOQ of 1.5 and 10 ng/mL(EGCG)as well as 0.75 and 5 ng/mL(ECG,EGC,and EC).A good linearity was obtained over a wide range of 10-10000 ng/mL for EGCG and 5-5000 ng/mL for other three catechins(r > 0.996).The method was validated to be reproducible and reliable,as evidenced by intra-batch and inter-batch precision of less than 10% and 11%,accuracy of 97.13%-106.05% and 99.22%-103.14%,respectively.The recovery of extraction ranged from 72.74% to89.13%,matrix effect from 88.76% to 105.97% for four cateckins.The method was successfully used to study the pharmacokinetics of TP iv administered to rats at a dose of 100 mg/kg.Conclusion This method is shown to completely meet requirements for the multi-component pharmacokinetic study of TP in rats.

OBJECTIVE:To investigate the neuropotective effect and anticoagulation effect of total Saponins of Radix Liriopes on focal cerebral ischemia in rats.METHODS:Chemical reagent Fecl3 was locally applied on the injured vessels to establish middle cerebral artery thrombosis model,and the effects of total Saponins of Radix Liriopes on rats' behavioral disturbance,brain infarct size,the histopathological changes in brain and the expression rate of nNOS immunoreactive positive neurons were measured,and the bleeding time and coagulation time were also detected with glass tube method and tail transection method.RESULTS:Due to the use of total Saponin of Radix Liriopes(10 and 40mg? kg-1),the brain infarct size was significantly decreased,the behavioral disturbance were improved and the expression rate of the nNOS immunoreactive positive neurons in rats were decreased.At doses of 20 and 60mg? kg-1,total Saponin of Radix Liriopes significantly prolonged the coagulation time and bleeding time.CONCLUSION:Total Saponin of Radix Liriopes has nuroprotective effect on focal cerebral ischemia induced by middle cerebral artery thrombosis and significant anticoagulation effect.

Arg-Gly-Asp (RGD) is a unique minimal integrin-binding sequence found within several glycoprotein ligands and also in snake-venom disintegrins. Adinbitor, a protein with 73 amino acid residues including 12 cysteins and an RGD motif, was cloned from Gloydius blomhoffi brevicaudus in my laboratory. As a new member of disintegrin family, adinbitor can inhibit both human platelet aggregation induced by ADP and angiogenensis in vivo and in vitro, the typical characters of disintegrin family. To separate the effect of inhibiting platelet aggregation from that of inhibiting angiogenensis, the motif KGD was introduced into adinbitor cDNA to replace RGD by site-directed and PCR-based mutagenesis. The recombinant protein (recombinant adinbitor (KGD)) was expressed in E. coli BL21 and purified through the His· Bind affinity chromatography. Recombinant adinbitor (KGD) could inhibit ADP-induced platelet aggregation with IC50 value of 85 nmol/L. Considerably, it was a more effective inhibitor on platelet aggregation than recombinant adinbitor (RGD), which has an IC50 of 150 nmol/L. Interestingly, recombinant adinbitor (KGD) has no potency in inhibiting angiogenesis in vivo compared with recombinant adinbitor (RGD). These findings showed that KGD containing adinbitor was more suitable for inhibiting ADP-induced human platelet aggregation as a potential and specific inhibitor of human platelet aggregation, which might have promising therapeutic potential as an antithrombotic agent.