molecules Article
Pharmacokinetic Comparison of Scutellarin and Paeoniflorin in Sham-Operated and Middle Cerebral Artery Occlusion Ischemia and Reperfusion Injury Rats after Intravenous Administration of Xin-Shao Formula Yueting Li 1,2 , Yuan Lu 1 , Jianchun Hu 1,3 , Zipeng Gong 1 , Wu Yang 1,3 , Aimin Wang 4 , Jiang Zheng 1 , Ting Liu 1 , Tingting Chen 1,3 , Jie Hu 1,3 , Ling Mi 1,3 , Yongjun Li 4 , Yanyu Lan 4 and Yonglin Wang 1, * 1
2 3 4
*
Key Laboratory of Pharmaceutics of Guizhou Province, Guizhou Medical University, No.9, Beijing Road, Yunyan District, Guiyang 550004, China;
[email protected] (Y.L.),
[email protected] (Y.L.),
[email protected] (J.H.),
[email protected] (Z.G.),
[email protected] (W.Y.),
[email protected] (J.Z.),
[email protected] (T.L.),
[email protected] (T.C.),
[email protected] (J.H.),
[email protected] (L.M.) National Engineering Research Center of Miao’s Medicines, Guiyang 550004, China School of Pharmacy, Guizhou Medical University, No. 9, Beijing Road, Yunyan District, Guiyang 550004, China Engineering Research Center for the Development and Applications of Ethnic Medicines and Traditional Chinese Medicine (TCM), Ministry of Education, Guizhou Medical University, No. 9, Beijing Road, Yunyan District, Guiyang 550004, China;
[email protected] (A.W.);
[email protected] (Y.L.);
[email protected] (Y.L.) Correspondence:
[email protected]; Tel.: +86-851-869-8468
Academic Editor: Jianbo Xiao Received: 24 July 2016; Accepted: 29 August 2016; Published: 7 September 2016
Abstract: Xin-Shao formula is a folk remedy widely used in China to prevent and cure stroke. Cerebral ischemic reperfusion (I/R) injury often takes place during the treatment of stroke. Information about the pharmacokinetic behavior of the remedy under cerebral I/R injury conditions is lacking. The present study aimed to compare the pharmacokinetic properties of scutellarin and paeoniflorin, two major bioactive components of Xin-Shao formula, under physiological state in cerebral I/R injury rats. Neurobehavioral dysfunction was evaluated and cerebral infarcted volume was measured in middle cerebral artery occlusion I/R injury (MCAO) rats. Plasma samples were collected at various time points after a single dose (intravenous, i.v.) of Xin-Shao formula. The levels of plasma scutellarin and paeoniflorin at the designed time points were determined by a UPLC-MS/MS method, and drug concentration versus time plots were constructed to estimate pharmacokinetic parameters. Increase in terminal elimination half-life (t1/2z ) and mean residence time (MRT(0–t) ) of scutellarin as well as elevation in area under the plasma drug concentration-time curve from 0 h to the terminal time point (AUC(0–t) ) and maximum plasma drug concentration (Cmax ) of paeoniflorin, along with decreased clearance of paeoniflorin and scutellarin as well as reduced apparent volume of distribution (Vz ) of paeoniflorin, were observed in MCAO rats, compared with those in sham-operated animals. The elimination of scutellarin and paeoniflorin were reduced in cerebral I/R injury reduced rats. Keywords: Xin-Shao formula; scutellarin; paeoniflorin; MCAO; pharmacokinetics; UPLC-MS/MS
Molecules 2016, 21, 1191; doi:10.3390/molecules21091191
www.mdpi.com/journal/molecules
Molecules 2016, 21, 1191 Molecules2016, 21, 1191
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Introduction 1. Introduction definedas asaasudden sudden neurological dysfunction in the brain caused by either ischemia Stroke, defined neurological dysfunction in the brain caused by either ischemia (lack (lack of blood supply to the brain) or a hemorrhage, has become one of the leading causes of severe of blood supply to the brain) or a hemorrhage, has become one of the leading causes of Around 80% 80% of of strokes result from neuronal impairment, loss of brain functions, and death [1]. Around Reperfusion is is considered considered as an effective approach cerebrovascular system blockage (ischemia) [2]. Reperfusion to improve the ischemic condition of the brain, particularly for vascular blockage, however, cerebral may induce induce an an accumulation accumulation of of reactive reactive oxygen oxygen species species released released from ischemia and reperfusion reperfusion (I/R) (I/R) may mitochondria, which which possibly possibly accelerates accelerates lipid lipid peroxidation peroxidation and and the the process process of of neuronal neuronal death death [3]. [3]. mitochondria, The resulting oxidative stress could worsen the extent of stroke [4]. could worsen the extent of stroke [4]. traditional Chinese Chinese medicines medicines (TCMs) (TCMs) are are reported reported to to facilitate facilitate recovery recovery from from stroke stroke [4]. [4]. Many traditional Xin-Shao formula, consisting consisting of Erigeron breviscapus breviscapus (Vant.) Hand.-Mazz Hand.-Mazz and and Paeonia Paeonia veitchii veitchii Lynch, Lynch, common TCM TCMand andhas hasbeen beenused used prevention treatment of acute infarcts is aa common forfor thethe prevention andand treatment of acute brainbrain infarcts and and stroke [5–7]. early studies demonstrated thatXin-Shao Xin-Shaoformula formularemarkably remarkably inhibited inhibited the stroke [5–7]. OurOur early studies demonstrated that arteriovenous bypass bypass thrombogenesis thrombogenesis in in rats [8], protected against cerebral I/R injury in in rats rats [9], [9], arteriovenous I/R injury microcirculation disturbance disturbance resulting resulting from fromblood blooddeficiency deficiencyin inischemic ischemicsite siteof ofbrain brain[9] [9],, improved microcirculation (MCAO) rats [9] and in and decreased reactive oxygen species in middle cerebral artery occlusion (MCAO) cultured PC12 cells [10]. pharmacokineticstudy study of Xin-Shao formula has performed been performed in rats normal in our A pharmacokinetic of Xin-Shao formula has been in normal in ourrats laboratory laboratory [11].the However, the pharmacokinetic behaviors of the herbal are medicine are not necessarily [11]. However, pharmacokinetic behaviors of the herbal medicine not necessarily the same the same under pathophysiological specifically in I/R cerebral I/RCerebral injury. Cerebral under pathophysiological conditions,conditions, specifically in cerebral injury. I/R injuryI/R is injury is commonly a major factor which could be antoobstacle effective cure commonly recognizedrecognized as a majorasfactor which could be an obstacle effectivetocure of stroke. of stroke.I/R Cerebral I/R injury wastoreported to alter pharmacokinetic behaviors of some herbal Cerebral injury was reported alter pharmacokinetic behaviors of some herbal medicines medicinesThe [2,12,13]. The primaryofobjective of was this study was topharmacokinetic compare pharmacokinetic of [2,12,13]. primary objective this study to compare propertiesproperties of Xin-Shao Xin-Shao formula in cerebral I/R injury rats and sham-operated rats. Scutellarin (Figure1A) 1A) and formula in cerebral I/R injury modelmodel rats and sham-operated rats. Scutellarin (Figure paeoniflorin (Figure 1B), two major components known injury [14–20], [14–20], known to protect protect against against cerebral cerebral I/R I/R injury were monitored thethe pharmacokinetic study.study. We hoped studythe would provide useful information monitoredforfor pharmacokinetic We the hoped study would provide useful to guide the to effective use effective of Xin-Shao in the clinic. in the clinic. information guide the use formula of Xin-Shao formula
Figure (B); and puerarin (C, (C, IS). IS). Figure 1. 1. The The structures structures of of scutellarin scutellarin (A); (A); paeoniflorin paeoniflorin (B); and puerarin
2. Results Results 2.1. Neurobehavioral Abnormality and Cerebral Infarcted Volume The neurological damage induced in rat model was evaluated by monitoring monitoring neurobehavioral dysfunction, using usingthe the five point scale. degree of neurobehavioral dysfunction in dysfunction, five point scale. The The degree of neurobehavioral dysfunction varied invaried MCAOMCAO-induced rats from one to another. Some of the MCAO-induced rats failed to fully flex the right induced rats from one to another. Some of the MCAO-induced rats failed to fully flex the forepaw, circled 0.01) forepaw, circled counter counter clockwise, clockwise,or orleaned leanedto tothe theleft leftside. side.The Thescores scoreswere were00vs. vs.2.2 2.2± ± 0.5 (p
