Xingnaojing Research Articles

Protective effect of Xingnaojing injection on ferroptosis after cerebral ischemia injury in MCAO rats and SH-SY5Y cells

Xingnaojing（XNJ）injection is a traditional Chinese medicine injection with neuroprotective effect, which has been widely used in the treatment of stroke for many years. This study aimed to explore the potential mechanism of XNJ in cerebral ischemia mediated by ferroptosis using proteomics and in vivo and in vitro experiments. After the rat model of middle cerebral artery occlusion (MCAO) was successfully established, they were randomly divided into model, XNJ, and deferoxamine (DFO) group. Triphenyl tetrazolium chloride (TTC) staining, Hematoxylin and eosin (H&E), and Nissl staining were used to observe the infarct area, pathological changes and the degree of neuronal apoptosis of rat brain. Proteins extracted from rat brain tissues were analyzed by quantitative proteomics using tandem mass tags (TMT). Western blotting and immunohistochemical assessment were used to measure the expression of ferroptosis-related proteins. In vitro, the SH-SY5Y cells were subjected to hypoxia (37°C/5% CO2/1% O2) for 24h to observe the survival rate, and detect the reactive oxygen species (ROS) content and ferroptosis-related proteins. In TTC and H&E experiments, we found that XNJ drug treatment reduced the infarct volume and brain tissue damage in MCAO rats. Nissl staining also showed that compared with MCAO group rats, the Nissl bodies of brain tissue after XNJ drug intervention were clear with a 3.54-fold increased times, suggesting that XNJ improved cerebral infraction, and neurological deficits in MCAO rats. Proteomics identified 101 intersected differentially expressed proteins (DEPs). According to the bioinformatics analysis, these DEPs were closely related to ferroptosis. Further research indicated that MCAO-induced cerebral ischemia was alleviated by upregulating recombinant glutathione peroxidase 4 (GPX4), ferroportin (FPN) expression, Heme oxygenase-1 (HO-1) expression, and downregulating cyclooxygenase-2 (COX-2), transferring receptor (TFR) and divalent metal transporter-1 (DMT1) expression after XNJ treatment. In addition, in vitro experiment indicated that XNJ improved the survival rate of hypoxia-damaged SH-SY5Y cells. XNJ increased the level of GPX4 and inhibited the protein expression of COX-2 and TFR after cell hypoxia. Moreover, different concentrations of XNJ (0.25%, 0.5%, 1%) reduced the ROS content of hypoxic cells, suggesting that XNJ could inhibit hypoxia-induced cell damage by regulating the expression of ferroptosis-related proteins and decreasing the production of ROS. XNJ could promote the recovery of neurological function in MCAO rats and hypoxia SH-SY5Y cells by regulating ferroptosis.

Studies of Mucosal Irritation and Cellular Uptake Mechanisms of Xingnaojing Nanoemulsion

Xingnaojing (XNJ) injection was used to treat pneumonia and stroke in clinic in China, but with poor patient compliance. Xingnaojing nanoemulsion for intranasal delivery was developed to improve it. This article tried to evaluate the mucosal irritation of Xingnaojing nanoemulsion and investigate cellular uptake mechanism of its encapsulated lipophilic drugs. The toad palate model and rat nasal mucosa model were used to study the nasal ciliotoxicity and nasal mucosal irritation of nanoemulsion to evaluate its safety intranasally. The cellular uptake mechanism was studied by Calu-3 cell model. Coumarin 6 was encapsulated in nanoemulsion and the endocytic pathways were studied by cellular uptake experiments after being treated with different inhibitors. In toad palate model, the cilia movement of Xingnaojing nanoemulsion group last for 467.40 ± 39.02 min, which was obviously longer than deoxycholate group (90.60 ± 15.40 min). Studies on rats showed that the damage caused by nanemulsion is capable of being recovered. Nanoemulsion uptake was reduced obviously when cells were treated with wortmannin, and it also decreased about 13% when the temperature reduced from 37ºC to 4ºC. Mucosal irritation caused by nanoemulsion is low and the damage is recoverable. The cellular uptake of Xingnaojing nanoemulsion is energy-dependent, and macropinocytosis was the most important pathway for cellular uptake.

Effects of Yunanan Baiyao adjunct therapy on postoperative recovery and clinical prognosis of patients with traumatic brain injury: A randomized controlled trial

BackgroundEffective therapies are needed to prevent the secondary injury and poor prognosis associated with emergency craniotomy of traumatic brain injury (TBI). Hypothesis/PurposeThe wound-healing medicine Yunnan Baiyao (YB) and Xingnaojing (XNJ) adjunct-therapy may improve the outcome of orthodox mono-therapy (OT). Study designRandomized controlled trial. MethodsEighty patients with moderate-to-severe TBI received emergency craniotomy (within 12 h after TBI) at the Chinese PLA General Hospital before being randomly assigned to 4 different treatments (n = 20) for 7 days: 1) OT; 2) OT+XNJ (i.v. 20 ml/daily); 3) OT+low dose-YB (oral, 1,000 mg/day); 4) OT+high dose-YB, 2,000 mg/day). ResultsGCS score was improved more quickly and became significantly higher in XNJ, l-YB, h-YB groups than in OT group (p<0.01). Serum S100B peaked higher but declined more slowly in OT group than in other groups (p<0.01). On postoperative Day 7, S100B was 20% below baseline in YB and XNJ groups but remained 19% above baseline in OT group which also lost 38% of superoxide dismutase (SOD) activity on Day 3 and recovered 69% of SOD on Day 7 whereas the YB and XNJ groups lost 16%∼23% of SOD activity on Day 3 and recovered 92%∼99% of SOD on Day 7 (p<0.01). Clinical prognosis (Glasgow Outcome Scale and Karnofsky Performance Scale) were significantly better (25%∼30%) in the XNJ, l-YB and h-YB groups than in OT group 3 months post-surgery and were correlated with serum S100B and SOD. ConclusionsYB and XNJ adjunct therapies improved postoperative recovery and clinical prognosis in patients with moderate-to-severe TBI partly through divergent regulation of S100B and SOD pathways. (The trial was registered at Chinese Clinical Trial Registry (ChiCTR) trial registration number: ChiCTR2000030280)

Effects of Xingnaojing on serum high-sensitivity C-reactive protein and neuron-specific enolase in patients with acute cerebral hemorrhage: A protocol of systematic review and meta-analysis.

Background:This study will systematically explore the effects of Xingnaojing (XNJ) on serum high-sensitivity C-reactive protein (hs-CRP) and neuron-specific enolase (NSE) in patients with acute cerebral hemorrhage (ACH).Methods:We will comprehensively search the following electronic databases (MEDLINE, EMBASE, Cochrane Library, Allied and Complementary Medicine Database, and China National Knowledge Infrastructure) from inception to the March 1, 2020. There are no limitations related to the language and publication status. Two authors will independently perform all citation identification, information extraction, and study quality. All potential conflicts will be solved through discussion with the help of a third author. RevMan 5.3 software will be used for data synthesis and statistical analysis.Results:This study will summarize the present evidence to investigate the effects of XNJ on serum hs-CRP and NSE in patients with ACH.Conclusion:This study may provide an impressive understanding of perspective from scientific basis for effects of XNJ on serum hs-CRP and NSE in patients with ACH.Study registration:PROSPERO CRD42020171648.

Xingnaojing ameliorates synaptic plasticity and memory deficits in an Aβ1–42 induced mouse model of Alzheimer's disease

The accumulation of insoluble amyloid β (Aβ) peptides is one of the pathological changes in Alzheimer's disease (AD), which induced synaptic plasticity impairment and excitatory amino acid toxicity associated with decreased memory function. Xingnaojing (XNJ), a well-known prescription in traditional Chinese medicine, has been used for the treatment of stroke for many years in China. In this study, we aim to investigate the therapeutic effects of XNJ in a hippocampus of Aβ1–42 induced mouse model of AD which showed significant memory loss and impaired synaptic morphology and function. Treatment of XNJ could attenuate spatial and working memory dysfunction, increase dendritic spine density and improve long-term potential (LTP) induction. In addition, XNJ treatment significantly increased the level of N-methyl-d-aspartate receptors (NMDARs) and inhibit the NMDA/α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) ratio in AD mice. XNJ treatment also activated the AKT/mechanistic target of rapamycin (mTOR) pathway, while inhibition of the mTOR pathway by rapamycin could reverse the protective effects of XNJ treatment. In conclusion, XNJ protected against synaptic plasticity and memory impairment in AD mice via the activation of AKT/mTOR signaling pathway, suggesting XNJ as an alternative treatment for AD.

Integrated analysis and network pharmacology approaches to explore key genes of Xingnaojing for treatment of Alzheimer's disease.

BackgroundAlzheimer's disease (AD), as a neurodegenerative condition, is one of the leading causes of dementia. Our study aims to explore the key genes of Xingnaojing (XNJ) for treatment of AD by integrated microarray analysis and network pharmacology.MethodsThe differentially expressed genes (DEGs) were identified in AD compared with normal control. According to these DEGs, we performed the functional annotation, protein–protein interaction (PPI) network construction. The network pharmacology was used to explore the potential targets of XNJ in the treatment of AD. The expression level of selected candidate genes was validated by quantitative real‐time polymerase chain reaction (qRT‐PCR).ResultsA total of 1,424 DEGs (620 genes were upregulated and 804 genes were downregulated) between AD and normal control were obtained. The functional annotation results displayed that neuroactive ligand–receptor interaction, regulation of actin cytoskeleton, Estrogen signaling pathway and notch signaling pathway were significantly enriched pathways in AD. Comparing the target genes of four active ingredients, a total of 16 shared genes were found. Among which, HTR2A and ADRA2A were also enriched in pathway of neuroactive ligand–receptor interaction. The expression of 4 DEGs (SORCS3, HTR2A, NEFL, and TAC1) was validated by qRT‐PCR. Except for TAC1, the other 3 DEGs in AD were consistent with our integrated analysis.ConclusionsThe results of this study may provide novel insights into the molecular mechanisms of AD and indicate potential therapeutic targets for AD.

Xingnaojing injection improves Aβ1-42-induced memory deficit in mice by altering of excitatory amino acid toxicity and synaptic plasticity

Alzheimer’s disease (AD), the widespread type of dementia, is characterized by depositions of senile plaques composed of insoluble amyloid β (Aβ) peptides, and neurofibrillary tangles. It causes progressive degeneration of synaptic plasticity and leads to memory loss and cognition impairment. Abnormal changes in synaptic structure and excitatory amino acid toxicity occurred in early stage in AD and are associated with decreased cognitive function. Xingnaojing (XNJ), a well-known prescription in traditional Chinese medicine, has been used for treatment of stroke in China. In this research, mice were randomly divided into 4 groups: control, Aβ1-42 injected (Aβ) and Aβ1-42 injected with two doses of XNJ (low-dose, high-dose) administration groups. We performed bilateral intra-CA1 injection of Aβ1-42 except control group. Behavioral results showed that the mice treatment with high-dose of XNJ group has a longer exploration time for New object recognition (NOR) and more crossing platform times for Morris water maze (MWM) compared with Aβ group. The expression of MAP-2 was increased by administration of XNJ in immunofluorescence results. Golgi staining showed that XNJ can improve dendritic spine density of damaged neurons. It also increased the protein levels of GAP-43, PSD-95, NR2b, p-AKT/AKT and p-mTOR/mTOR in the brain tissues compared with Aβ group in the Western blot (WB). Our results indicated that XNJ exhibited a protective effect against excitatory amino acid toxicity and synaptic plasticity via AKT/mTOR signal pathway in mice with Aβ1-42-induced memory deficit. These results provided evidences for the novel and potential application of XNJ for the treatment of AD.

Evaluation of the adjunctive effect of Xing Nao Jing Injection for viral encephalitis: A systematic review and meta-analysis of randomized controlled trials.

Background:To systematically evaluate the effect and safety of Xing Nao Jing (XNJ) injection as an add-on treatment on the treatment for viral encephalitis (VE).Methods:Trials assessing the adjunctive effectiveness of XNJ injection for VE were searched from 4 electronic databases from inception to October 31, 2018. Two authors independently extracted data and assessed risk of bias. Statistical analyses were performed using RevMan 5.3 software. Meta-analysis and additional analysis were conducted if data permitted. Trial Sequential Analysis and Grading of Recommendations Assessment, Development and Evaluation (GRADE) were also performed.Results:This review involved 23 trials and 1757 participants, all trials were assessed as having unclear risk of bias. Results from 5 meta-analyses, 13 subgroup meta-analyses, and the single studies showed that based on conventional therapy XNJ injection (0.4–0.6 mL/kg daily for children, 20 mL/day for adults) may have better effect on increasing the numbers of cured patients and decreasing the time of recovery of main symptoms for patients with viral encephalitis. Patients used combination of XNJ injection and conventional therapy had higher cured rate (risk ratio 1.61, 95% confidence interval 1.45–1.80, 19 trials, 1456 participants) and less mortality rate (risk ratio 0.26, 95% confidence interval 0.10–0.71, 9 trials, 595 participants). The average difference of time for fever, conscious, or convulsive recovery was average 2 hours shorter in combination group than in control. No difference was found between children and adults according to the subgroup analysis. Safety of the XNJ injection was failed to evaluate due to the insufficient evidence in this review.Conclusions:This review found “very low” quality evidence which showed the potential effectiveness of combination of XNJ injection and conventional therapies for VE. Considering the TSA results, conclusion could only be draw on effectiveness of the XNJ injection as add-on treatment for VE patients on increasing the cured rate. Firm conclusion on other outcome measures for effectiveness assessment or safety of XNJ injection could not be draw according to this review due to the insufficient evidence.

Xingnaojing Injection Protects against Cerebral Ischemia Reperfusion Injury via PI3K/Akt-Mediated eNOS Phosphorylation.

Xingnaojing (XNJ) injection, derived from traditional Chinese medicine formulation, has a protective effect against stroke, but the underlying mechanism is unclear, which severely limited its clinical application. This research aims to elucidate the role and mechanism of XNJ in reducing cerebral ischemic reperfusion (I/R) injury. Rats received 2 h cerebral ischemia followed by reperfusion of 24 h and were intraperitoneally given 5, 10, or 15 ml/kg XNJ 24 h before ischemia and at the onset of reperfusion, respectively. TTC staining, HE staining, and neurological score were implied to evaluate the effectiveness of XNJ. The protein expressions of PI3K/Akt and eNOS signaling were measured. Experiments were further performed in human brain microvascular endothelial cells (HBMECs) to investigate the protective mechanisms of XNJ. HBMECs were subjected to 3 h oxygen and glucose deprivation following 24 h of reoxygenation (OGD) to mimic cerebral I/R in vitro. PI3K inhibitor LY294002 was added with or without the preconditioning of XNJ. Multiple methods including western blot, immunofluorescence, DAPI staining, JC-1, and flow cytometry were carried out to evaluate the effect of XNJ on HBMECs. XNJ could improve rat cerebral ischemic injury and OGD induced HBMECs apoptosis. In vivo and in vitro researches indicated that the mechanism might be relevant to the activation of PI3K/Akt/eNOS signaling.

Systems pharmacology dissection of the anti-stroke mechanism for the Chinese traditional medicine Xing-Nao-Jing

Xing-Nao-Jing (XNJ) is a well-known injection that has been extensively applied in clinical treatment of stroke in China. However, the underlying mechanism of clinical administration of XNJ in stroke remains unclear. In this study, a systems pharmacology strategy based on pharmacokinetic and pharmacodynamics data was applied to analyze the pharmacological effect of XNJ on stroke. Sixteen active compounds were filtered from XNJ through Drug-likeness (DL) and Brain-blood-barrier (BBB) evaluations. Ninety-four potential targets of these active components were identified by SysDT and SEA. Biological process and pathway enrichment analyses of these targets demonstrated that XNJ exerted anti-stroke effects by biological processes and pathways, such as the response to oxidative stress, regulation of blood pressure, calcium signaling pathway, and apoptosis. Integrating the compound-target network and stroke-related PPI network, we found that Akt1, HIF-1α and ITGB2 may play key roles in the treatment of stroke. The experiments demonstrated that oxycurcumenol may prevent PC12 cells from oxidative stress-induced cell damage. Our study indicates that XNJ has an effect on stroke by protecting neuro cells from oxidative stress-induced cell damage via HIF1α, and the research strategy at the systems pharmacology level is feasible to reveal the mechanisms of novel lead compounds from natural products.

Meta-Analysis of the Effects of Xingnaojing Injection on Consciousness Disturbance.

Xingnaojing (XNJ) is commonly extracted from Angongniuhuang, a classic Chinese emergency prescription, and widely used in the treatment of nervous system disorders including consciousness disturbance in China.To evaluate the beneficial and adverse effects of XNJ injection, on consciousness disturbance.Seven major electronic databases were searched to retrieve randomized controlled trials designed to evaluate the clinical efficacy of XNJ alone or combined with Western medicine in treating consciousness disturbance caused by conditions such as high fever, poisoning, and stroke. The methodological quality of the included studies was assessed using criteria from the Cochrane Handbook for Systematic Review of Interventions, and analyzed using the RevMan 5.3.0 software.Seventeen randomized controlled trials on XNJ were included in this study and the trials generally showed low methodological quality. The results revealed that XNJ alone or in combination with other medicines and adjuvant methods had a positive effect on patients with fever-, poisoning-, and stroke-induced coma.XNJ effectively treated consciousness disturbances that were caused by high fever, poisoning, or stroke.

XingNaoJing, prescription of traditional Chinese medicine, prevents autophagy in experimental stroke by repressing p53-DRAM pathway

BackgroundXingnaojing (XNJ), a well known prescription in traditional Chinese medicine, has been used for treatment of stroke in China. However, the effects and mechanisms of XNJ on autophagy are not clear. Here, we used the cell models of autophagy induced by serum-free condition and ischemia stroke in rats to further investigate whether the p53-DRAM pathway is involved in the effects of XNJ on autophagy.MethodsWe used the cell model of autophagy induced by serum-free condition and the rat model of ischemia caused by a middle cerebral artery occlusion (MCAO). The effects of XNJ on p53 transcriptional activity of PC12 cells were evaluated by the luciferase activity assay. The mRNA levels and the expression of p53 and its target autophagy gene DRAM (damage-regulated autophagy modulator) were analyzed respectively by Quantitative-RTPCR and Western blot assay. The activation of autophagy was detected by the levels of autophagy markers, microtubule associated protein light chain 3 (LC3) and p62 by Immunofluorescence and Western blot. p53 inhibitor was used to determine whether p53 is responsible for the effects of XNJ on preventing autophagy.ResultsThe assay for luciferase activity of p53 promoter indicated that XNJ inhibited p53 transcriptional activity. XNJ reduced the expression of p53 and its target autophagy gene DRAM (damage-regulated autophagy modulator) in serum-free condition PC12 cells and the cortex in MCAO rats. XNJ reduced autophagy of PC12 cells induced by serum-free condition and the cortex in MCAO rats. Furthermore, suppression of p53 by p53 inhibitor significantly reduced the effects of XNJ on the autophagy of PC12 cells in serum-free condition.ConclusionXNJ prevents autophagy in experimental stroke by repressing p53/DRAM pathway. Our findings are therefore of considerable therapeutic significance and provide the novel and potential application of XNJ for the treatment of brain diseases.

Pre-treatment with a Xingnaojing preparation ameliorates sevoflurane-induced neuroapoptosis in the infant rat striatum.

Xingnaojing (XNJ), is a standardized Chinese herbal medicine product derived from An Gong Niu Huang Pill. It may be involved in neuroprotection in a number of neurological disorders. Exposure to anesthetic agents during the brain growth spurt may trigger widespread neuroapoptosis in the developing brain. Thus the present study aimed to identify whether there was a neuroprotective effect of XNJ on anesthesia-induced neuroapoptosis. Seven-day-old rats received treatment with 2.1% sevoflurane for 6 h. Rat pups were injected intraperitoneally with 1 or 10 ml/kg XNJ at 0.2, 24 and 48 h prior to sevoflurane exposure. The striata of neonatal rats were collected following administration of anesthesia. Western blotting and immunohistochemistry were used to analyze the expression of activated caspase 3, Bax and phosphorylated protein kinase B (p-AKT) in the striatum. It was found that activated caspase 3 and Bax expression were upregulated in the striatum following sevoflurane treatment. Preconditioning with XNJ attenuated the neuronal apoptosis induced by sevoflurane in a dose-dependent manner. Anesthesia reduced the expression of p-AKT (phosphorylated at sites Thr308 and Ser473) and phosphorylated extracellular-regulated protein kinase (p-ERK) in the striatum. Pre-treatment with XNJ reversed the reduction in p-AKT, but not p-ERK expression. These data suggest that XNJ has an antiapoptotic effect against sevoflurane-induced cell loss in the striatum. It thus holds promise as a safe and effective neuroprotective agent. The action of XNJ on p-AKT may make a significant contribution to its neuroprotective effect.

Influence of Xingnaojing Injection on the Expression of c-fos and c-jun Proteins in Brains of Rats in a Kindling Model of Epilepsy Chronically Induced by Pentetrazol

Objective: To investigate the molecular biological mechanism of the antiepileptic effect of Xingnaojing (XNJ) injection on a rat kindling model of epilepsy (KME). Methods: Fifty healthy 6-week-old male Sprague-Dawley rats were divided into the following five groups: blank control (BC) group, model (M) group, XNJ injection (XI) group, phenobarbital injection (PI) group and XNJ combined with phenobarbital (XP) group. There were 10 rats in each group. The intervention drugs were administered 30 min before the model-building drugs once a day for 5 weeks. The model-building drug pentetrazol was given to each group as an intraperitoneal injection 30 min after the use of the intervention drug once a day for 4 weeks for KME establishment (except for the BC group). The BC group was given physiological saline instead. All drugs were injected intraperitoneally. The behaviors of each group of rats were observed after the use of the model-building drugs for 1 h every day. The last kindling test was carried out at the end of week 5. Then, c-fos and c-jun protein expressions in the rat brains of each group were observed and analyzed by immunohistochemistry at the end of the experiment. Results: There was a large number of cells positive for the c-fos and c-jun proteins in the rat brains of the M group. Compared with the M group, the expression level of the c-fos and c-jun proteins was lower in the rat brains of the XI and PI groups (p < 0.01). There was no statistical difference between the XI and PI groups (p > 0.05). The number of positive cells in the rat brains of the XP group was even smaller than that of the XI or PI groups. Conclusion: The antiepileptic effect of the XNJ injection on the rat KME is probably related to its interruption function on the expression of the c-fos and c-jun proteins in rat brains.

Preparation of mPEG2000-PLA-modified Xingnaojing microemulsion and evaluation in mucosal irritation

Xingnaojing microemulsion (XNJ-M) administered intranasally is used for stroke treatment. Methoxy poly(ethylene glycol)-poly(lactide) (mPEG-PLA) is a block copolymer with surfactant-like properties and good biodegradability and reliable biological safety. In order to investigate the possibility to lower mucosa irritation with XNJ-M, XNJ-M modified by mPEG2000-PLA (XNJ-MM) was prepared. Different ratios of mPEG2000-PLA were synthesized. The structures and properties were confirmed by 1H-NMR, IR, and DSC. The hydrophile–lipophile balance (HLB) value and critical micellar concentration of copolymers were investigated. The in situ toad palate model was adopted to investigate the ciliotoxicity of the copolymers. The XNJ-M and XNJ-MM were prepared by dropping aqueous phase method. Mucosal irritation of different Xingnaojing (XNJ) preparations was studied by behavioral observations including sneezing and scratching nose. The epithelial thickness of nasal mucosa was evaluated and the secretory protein concentration was determined. The results of in situ toad palate model demonstrated that high HLB value mPEG2000-PLA basically showed no ciliotoxicity, while EL-35 had significant dose-dependent ciliotoxicity (P < 0.05). The irritating effects of mPEG2000-PLA were significantly lower than EL-35 after seven days of treatment, based on the epithelial thickness of 0.429 ± 0.100 mm and 0.700 ± 0.035 mm, respectively (P < 0.01). The epithelial thickness with XNJ-MM was lower than XNJ-M after seven days of treatment: 0.620 ± 0.10 mm vs. 0.809 ± 0.153 mm, respectively (P < 0.05). The mPEG2000-PLA-modified microemulsion is a promising dosage form of XNJ, based on reduced irritation on nasal mucosa.

Comparative pharmacokinetics of borneol in cerebral ischemia-reperfusion and sham-operated rats

This study was designed to investigate the pharmacokinetics of borneol in the pathological conditions of stroke and evaluate the pharmacokinetic differences of borneol caused by stroke after oral administration of borneol and Xingnaojing (XNJ). The rats were divided into two groups, ischemia-reperfusion (IR) and sham-operated (SO) rats. Each group contained two subgroups: pure borneol and XNJ subgroups. After administration with the same dosages of borneol 162.0 mg/kg, plasma samples were collected. The cerebral ischemia-reperfusion model was created by reversible middle cerebral artery occlusion (MCAO). The blood samples were collected punctually after oral administration and a specific gas chromatographic system-flame ionization detector (GC-FID) method was developed and employed to determine the level of borneol in the plasma. The pharmacokinetic parameters were analyzed using non-compartmental methods with Kinetica. After administration of borneol, the maximum plasma concentration (Cmax) and area under the curve (AUC) values in stroke rats significantly increased by 302% and 275%, respectively, compared with the SO rats, and the same phenomenon appeared after administration of XNJ. In the rats with the same physiological conditions, the Cmax and AUC had higher values in the borneol subgroup (P<0.05). These results suggest that the pathological damages of ischemia-reperfusion have a significant impact on the pharmacokinetic traits of borneol and that there are some components in XNJ inhibiting the absorption of borneol.

Comparative pharmacokinetic studies of borneol in brain and plasma of stroke or sham-operated rats after administration of Xingnaojing

In order to research the pharmacokinetic characteristic of borneol in plasma and brain of stroke rats given XNJ and investigate the influence of stroke on the borneol passing through the blood-brain barrier, this study established the GC to determine the borneol in brain and blood, and made the stroke mode rats by middle carotid artery occlusion (MCAO) and set sham-operated group. After oral administration of Xingnaojing (XNJ) suspension, their blood and brain were collected at different time and detected by GC. The data was analysed by Kinetica. Results showed that in stroke group, the Cmax and AUC0-t of brain and plasma are (1.82 +/- 0.825), (1.35 +/- 0.43) mg x L(-1) and (123.39 +/- 55.82), (87.91 +/- 39.81) mg x L(-1) x min, Te (brain/blood drug ratio) was 70.93%; those pharmacokinetic values were larger than in sham-operated group. We can conclude that the pathological state of stroke can increase the amount of borneol permeating into brain.

Comparative study on rat in situ nasal absorption of geniposide of Xingnaojing nasal drop and Xingnaojing microemulsion

Xingnaojing (XNJ) is an effective clinical drug used to treat acute stroke. Compared with injection administration, its nasal administration has better brain targeting. Therefore, through nasal administration, XNJ microemulsion could help solve the drug load of compound components of different polarities contained in large-dose and high-concentration traditional Chinese medicines, and reduce irritation to nasal mucosa In this study, the modified volume correction method and the improved rat in situ nasal perfusion model were adopted to compare the nasal absorption of geniposide contained in different XNJ preparations. The results showed that the constant absorption rate of geniposide (GE) in XNJ-D was (2.95 +/- 0.25) x 10(-3) min(-1), whereas the constant absorption rate of GE in XNJ-M was (2.16 +/- 0.21) x 10(-3) min(-1). This indicated that the rat nasal absorption of GE in different XNJ preparations complied with the first-order process and could be considered as passive absorption. GE in XNJ-D was absorbed faster than that in XNJ-M, which provided basis for the development of nasal preparations of XNJ.

Antioxidant Action of Solid Preparation of Xingnaojing in SHRSP

We investigated the antioxidant action of a solid preparation of Xingnaojing (XNJ) and ascorbic acid (AA) in stroke-prone spontaneously hypertensive rats (SHRSP). The total antioxidant states in the plasma, systolic blood pressure, and heart rate were measured every 2 weeks, and lipid peroxidation, expressed as thiobarbituric acid-reactive substances in plasma, was measured in the 6th week. The results showed that AA and XNJ significantly increased the total antioxidant status in plasma and reduced malondialdehyde in the plasma. These data suggest that during 6-week administration, XNJ has antioxidant action on SHRSP, which may relate to its generalized inhibition of lipid peroxidation and promotion of the total antioxidant state. These results demonstrated that orally treated XNJ has an antioxidant effect on SHRSP plasma.

Effect of Xingnaojing injection on hippocampal N-methyl-D-aspartic acid receptors of focal cerebral ischemia in rats

Objective: To observe and elucidate the neuroprotective effect of Xingnaojing (XNJ) injection on hippocampal N-methyl-D-aspartic acid (NMDA) receptors of focal cerebral ischemia in rats.Methods: Cerebral ischemia was established by occluding the middle cerebral artery with an intraluminal suture technique in rats. Neurological deficit score, infarct volume and quantity of NMDA receptors were estimated in all groups and compared.Results: After being treated with XNJ, the score decreased in the initial 6 hours and infarct volume decreased in 24 hours. And within 24 hours, the quantity of NMDA receptors obviously decreased compared with the model group (P<0. 01). It indicated that XNJ could ameliorate neurological behavior of middle cerebral artery occlusion rats and down-regulate the expression of hippocampal NMDA receptors.Conclusion: The neuroprotective effect of XNJ on focal cerebral ischemia is possibly related to down-regulating the expression of NMDA receptors in rats.