Effect of Xingnaojing Injection(醒脑静注射液)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.

Effects of Xingnaojing Injection on Cerebral Neurological Function and ET-1, hs-CRP and TNF-α Levels in Elderly Patients with Acute Cerebral Infarction

OBJECTIVE: To observe the effects of Xingnaojing Injection on the function of cranial nerves and the level of ET-1,hs-CRP and TNF-α in elderly patients with acute cerebral infarction. METHODS: A total of 86 elderly patients with acute cerebral infarction were randomly divided into observation group and control group. The 43 cases in the control group were treated by conventional Western medicine, and 43 cases in the observation group were treated by Xingnaojing injection on the basis of the treatment in control group. After 2 weeks' treatment in Xuancheng People's Hospital of Anhui Province, the neurological function(NIHSS score and MMSE score), daily living ability(BI Index score and ADL score),laboratory indicators(hs-CRP, ET-1, TNF-α) of the 2 groups before and after the treatment were observed and compared,the total clinical effectiveness of the 2 groups were compared. RESULTS: The total effective rate was 88.4% in the observation group, which was signi?cantly higher than that in the control group(62.8%)(P < 0.05). The NIHSS scores in the 2 groups were both signi?cantly increased after the treatment(P < 0.05) while the MMSE scores were signi?cantly decreased(P < 0.05) in the 2 groups, and the scores' improvement in the observation group was signi?cantly higher than that in the control group(P < 0.05). The BI score and ADL score were significantly increased in the 2 groups after the treatment(P < 0.05), and the improvements were obvious in the observation group. The levels of ET-1, hs-CRP and TNF-αin the 2 groups were signi?cantly decreased after the treatment(P < 0.05), and the indexes in the observation group were signi?cantly lower than that in the control group(P < 0.05). CONLUSIONS: Xingnaojing injection can reduce the levels of serum ET-1, hs-CRP and TNF-α, as well as the degree of neurological de?cit in the treatment of elderly patients with acute cerebral infarction. It is also helpful for the prognosis and outcomes of patients with cerebral infarction.

Effect of Xingnaojing injection on penumbra transformation in rats with acute cerebral ischemia

Objective:To observe the effect of Xingnaojing injection on penumbra transformation in rats with acute cerebral ischemia.Methods:The rat model of middle cerebral artery occlusion(Middle cerebral artery occlusion,MCAO)was established by suture occlusion.Except the sham operation group,the other groups were randomly divided into model group and Xingnaojing group.The rats in Xingnaojing group were intraperitoneally injected with Xingnaojing injectionaccording to 0.18ml/100g,and the sham operation group and model group were given the same amount of normal saline respectively.24 hours after the establishment of the model,the morphological changes of neurons in the penumbra of the rats were observed by Nissl staining,the ultrastructural changes of neurovascular unit(Neurovascular unit,NVU)were observed by transmission electron microscope(Transmission electron microscope,TEM),and the apoptosis of the ischemic penumbra was detected by in situ apoptosis(TdT-mediated Dutp Nick-End Labeling,TUNEL).Magnetic resonance imaging was used to observe the ischemic evolution of the penumbra of the same rat at 4.5 h and 24 h,respectively.Results:Compared with the sham operation group,the number of neurons in the model group was significantly reduced,the structure of Nissl corpuscles was destroyed,the outline was blurred or disappeared,the pathological morphology of NVU ultrastructure was obviously damaged under transmission electron microscope,a large number of apoptotic cells could be seen in the model group by TUNEL staining(P<0.01),and magnetic resonance imaging showed that there was a large area infarction in the brain tissue of the model group.Compared with the model group,the pathomorphology of neurons and NVU ultrastructure in Xingnaojing group was significantly improved,the number of apoptotic cells was significantly decreased(P<0.01),and the loss rate of penumbra was significantly lower in Xingnaojing group(P<0.05).Conclusion:Xingnaojing injection can improve the state of neurons in ischemic penumbra,reduce the injury of glial cells and microvessels,inhibit apoptosis,promote the transformation of penumbra in rats with acute cerebral ischemia,and save part of penumbra to some extent.it has a certain protective effect on the brain tissue of penumbra in the acute stage of cerebral ischemia.

Mechanism of Xingnaojing injection intervention in cerebral ischemiareperfusion rat model based on GC-MS metabolomics

Objective:To investigate the protective effect of Xingnaojing injection on cerebral ischemia-reperfusion in rats and its metabolic pathway and mechanism.Methods:The cerebral ischemia reperfusion model of rats was established by suture occlusion.After successful model evaluation,he rats were randomly divided into model group and Xingnaojing group with eight rats in each group.In the sham operation group,only blood vessel separation was performed without embolization.Xingnaojing group was given intraperitoneal injection,model group and sham operation group were given the same dose of normal saline,twice a day.Three days later,HE staining and GC-MS metabolomics were used to detect the changes of endogenous metabolites in the rat brain tissue.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to screen out differential metabolites and analyze their metabolic pathways.Results:Endogenous metabolites were disturbed after cerebral ischemia-reperfusion injury in rats.Seventy-one different metabolites were screened from the model group and the sham group,of which three were down-regulated and sixty-eight were up-regulated.Eighty-eight different metabolites were found between Xingnaojing group and sham operation group,among which eight were down-regulated and eighty up-regulated.After screening of Xingnaojing group and model group,twelve different metabolites were obtained,among which seven were down-regulated and five up-regulated.By analyzing the differences of metabolites,Xingnaojing injection was considered to be involved in the metabolic pathway after cerebral ischemia-reperfusion in rats,including amino acid metabolism(beta alanine metabolism,alanine,glutamic acid and aspartic acid metabolism,histidine metabolism,arginine and proline metabolism),glutathione metabolism,pyrimidine metabolism,ABC transporter,nitrogen metabolism and other metabolic pathways.Conclusion:Xingnaojing injection can restore the levels of metabolites in cerebral ischemia-reperfusion rats in certain degrees,mainly through amino acid metabolism,ABC transporter,glutathione metabolism and other metabolic pathways to regulate cerebral ischemia-reperfusion injury in rats.

Clinical Study of Xingnaojing Injection Combined with Craniocerebral Hypothermia Apparatus in the Treatment of Acute Hemorrhagic Stroke

Objective:To investigate the clinical effect of Xingnaojing injection combined with craniocerebral hypothermia apparatus in the treatment of acute hemorrhagic stroke.Methods:Seventy-two patients with acute hemorrhagic stroke admitted to our hospital from March 2018 to September 2019 were randomly divided into two groups,with 36 cases in each group.The control group(n=36)was treated with craniocerebral hypothermia apparatus,and the observation group(n=36)was treated with Xingnaojing injection combined with craniocerebral hypothermia apparatus.The clinical efficacy,the National Institutes of Health Stroke Scale(NIHSS)score and the activity of daily living(MBI score)after 1 month of treatment were compared.Results:The effective rate of treatment in the observation group was higher than that in the control group(P<0.05);NIHSS score was decreased in both groups(P<0.05);MBI score was increased in both groups,and the change range of the observation group was greater than that of the control group,showing statistically significant differences(P<0.05).Conclusion:Xingnaojing injection combined with craniocerebral hypothermia apparatus can improve the clinical efficacy,the neurological impairment and activity of daily living in the treatment of acute hemorrhagic stroke,which is worthy of clinical application.

Effect of minimally invasive evacuation of hematoma combined with Xingnaojing therapy on neurological function injury and cytokine level in patients with hypertensive cerebral hemorrhagen

Objective:To study the effect of minimally invasive evacuation of hematoma combined with Xingnaojing therapy on neurological function damage and cytokine level in patients with hypertensive cerebral hemorrhage.Methods:A total of 80 patients with hypertensive cerebral hemorrhage treated in our hospital between June 2010 and September 2015 were selected as the research subjects, the treatment methods and test results were reviewed, and then they were divided into the control group (n=45) who accepted minimally invasive evacuation of hematoma alone and the observation group (n=35) who accepted minimally invasive evacuation of hematoma combined with Xingnaojing therapy. Before and after treatment, cerebral blood flow detector was used to detect cerebral blood flow parameters;ELISA method was used to detect serum levels of neurological function indexes and inflammatory cytokines;high performance liquid chromatograph was used to detect serum neurotransmitter levels. Results: Before treatment, the differences in cerebral blood flow parameters, neurological function indexes, inflammatory cytokines and neurotransmitters were not statistically significant between two groups of patients. After treatment, cerebral blood flow parameters Q and V levels of observation group were higher than those of control group while R level was lower than that of control group;serum NSE, NPY, IL-1β, IL-4, IL-6, TNF-α, Glu and Asp contents of observation group were lower than those of control group while BDNF and Gly contents were higher than those of control group.Conclusion: Minimally invasive evacuation of hematoma combined with Xingnaojing therapy can improve the neurological function and regulate the synthesis of inflammatory cytokines and neurotransmitters in patients with hypertension cerebral hemorrhage.

Effect of Xingnaojing Injection on inflammatory cytokines and blood coagulation function in patients with acute cerebral hemorrhage

Objective: To investigate the effect of Xingnaojing Injection on inflammatory cytokines and blood coagulation function in patients with acute cerebral hemorrhage. Method: A total of 80 patients with acute cerebral hemorrhage admitted to our hospital from October 2015 to May 2017 were randomly divided into the observation group and the control group, each group with 40 cases. Two groups both received conventional treatment;the observation group was given Xingnaojing injection additionally. The two groups were treated for two weeks. Changes of serum inflammatory factors TNF-α, hs-CRP, IL-10 and blood coagulation indexes of activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (Fib), thrombin time (TT) and D-dimer (DD) of two groups were compared. Results: Before treatment, the differences of inflammatory factors TNF-α, hs-CRP, IL-10 and coagulation function indexes APTT, PT, Fib, TT and DD of the two groups were not statistically significant. After treatment, the levels of TNF-α, hs-CRP of the two groups were significantly decreased, and the IL-10 level of the two groups were significantly increased;the levels of TNF-α, hs-CRP in observation group were significantly lower than the control group, and the IL-10 level in observation group were significantly higher than the control group. After treatment, the levels of APTT, Fib and DD in the control group were significantly increased;the levels of PT and DD in the observation group were significantly decreased and the Fib level was significantly increased. After treatment, the levels of APTT, PT and DD in the observation group were significantly lower than those in the control group, and Fib level was significantly higher than that in the control group, these differences were statistically significant. There was no significant difference of TT levels between the two groups before and after the treatment. Conclusion: Xingnaojing injection treatment of acute cerebral hemorrhage can significantly reduce the level of serum inflammatory factors and promote the restoration of coagulation function. It is worth promoting.

Clinical Study on the Treatment Efficacy of Cerebral Hemorrhage with Xingnaojing Injection in Real World

Objective:The purpose of this study was to understand the clinical efficacy of Xingnaojing Injection in the treatment of cerebral hemorrhage in the real world.Methods:The hospitalized patients with cerebral hemorrhage mainly diagnosed in the hospital information system(HIS)of 223 A hospitals in the country were selected as the research subjects.The patients with cerebral hemorrhage using Xingnaojing Injection were used as the Xingnaojing group.The group was matched with the same sex,age±5 years old,similar hospital admission conditions,matched according to 1:1 ratio.The patients with cerebral hemorrhage without using Xingnaojing Injection were used as the control group,and the therapeutic effects after treatment were used as outcome evaluation index.A total of 3 Logistic regression methods were used to analyze the effects of Xingnaojing Injection in the treatment outcome of patients with cerebral hemorrhage.Results:The statistical results showed that there was a difference in the efficacy of the treatment of cerebral hemorrhage by the use of Xingnaojing Injection(P<0.05).Conclusion:Based on the existing analytical methods and data,the effects of Xingnaojing Injection on cerebral hemorrhage are better than that of the non-use control group,which provides reference for the clinical drug use of cerebral hemorrhage.

Anticonvulsant Effect of Xingnaojing Injection on Acute Seizure Models in Mice

Background:Epilepsy is characterized by acute recurrent seizures.The control of seizures is largely hampered by the tolerance to current anti-seizure drugs.Complementary anti-convulsant pharmacotherapies are urgently needed.Objective:Here,we aimed to investigate the anti-convulsant effects of Xingnaojing Injection(XNJ)which is an approved Traditional Chinese Medicine injection on different acute seizure models in mice.Methods:The effects of XNJ were tested on the maximal electroshock(MES),pentylenetetrazol(PTZ)and kainic acid(KA)acute seizure models.Also,whether XNJ can directly inhibit hippocampal neuronal firings were exam-ined by in vitro electrophysiology.Results:XNJ could shorten the durations of generalized tonic-clonic seizures in the MES model.It also significantly prolonged the latencies to generalized myo-clonic seizures in the PTZ model.In the KA model,XNJ showed various efficacies including inhibiting the seizure stages,prolonging the latency to the occurrence of the first seizures or generalized seizures,shortening the seizure durations,decreasing the numbers of generalized seizures.In vitro electrophysiological recordings further verified XNJ directly inhibited both the spontaneous and evoked action potentials of hippocampal pyramidal neurons,but did not influence the excitatory or inhibitory synaptic transmissions.Conclusion:These findings proposed XNJ as an alternative anti-convulsant pharmacotherapy for controlling acute epileptic seizures.

XingNaoJing injections protect against cerebral ischemia/reperfusion injury and alleviate blood-brain barrier disruption in rats, through an underlying mechanism of NLRP3 inflammasomes suppression

The aim of this study was to explore the neuroprotective effect and mechanism of XingNaoJing injections(XNJ) on cerebral ischemia injury and blood-brain barrier(BBB) disruption. Middle cerebral artery occlusion(MCAO) method was applicated to establish the model of cerebral ischemia/reperfusion(I/R) injury in rats. BBB permeability after I/R injury was assessed with the leaking amount of Evans Blue and the expression of occludin and ZO-1. The expression of NOD-like receptor family, pyrin domain containing(NLRP3) was checked to explore the inhibition of inflammation by XNJ. The results showed that XNJ could significantly increase the survival percent, decrease the infarct area and ameliorate neurological deficits and brain damage after I/R injury. Leaking amount of Evans Blue was reduced by XNJ, and the expression of tight junction protein, occludin and ZO-1 was also up-regulated by XNJ, which showed a role of protection on BBB disruption. The expression of NLRP3 was inhibited after exposure of XNJ, which was associated with inhibition of the inflammatory response. In summary, XNJ could suppress NLRP3 inflammasomes and improve BBB disruption and brain damage in rats after cerebral I/R injury, which provided a beneficial insight to further explore XNJ.

Comparison between Xingnaojing Injection(醒脑静注射液)and Naloxone in Treatment of Acute Alcohol Intoxication:An Updated Systematic Review and Meta-Analysis of Randomized Controlled Trials

Objectives:To investigate the effectiveness and safety of Xingnaojing Injection(XNJ,醒脑静注射液)compared with naloxone for the treatment of acute alcohol intoxication(AAI),and provide the latest evidence through evidence-based approach.Methods:Seven electro-databases including Pub Med,EMBASE,Cochrane Central Register of Controlled Trials,Chinese National Knowledge Infrastructure Databases,Chinese Biomedical Literature Database,Chinese Science and Technology Periodical Database(VIP)and Wanfang Database were searched from the inception to January 2018.Randomized controlled trials(RCTs)comparing XNJ with naloxone for patients with AAI and reporting at least one of the below outcomes were included:patients’conscious recovery time,stay length in emergency department,disappearance time of the ataxia symptom,the severity of the symptoms,the blood alcohol content as wel as the adverse events.Methodological quality of included trials was assessed using the risk of bias tool which recommended by the Cochrane Col aboration.Meta-analysis was conducted by Review Manager 5.3 software.Results:Total y 141 trials with 13,901 patients were included in this review,al of them were assessed as unclear or high risk of bias.Results showed that on the basis of routine therapy,standard dose XNJ(10–20 m L)may have similar results with naloxone on the recovery time of consciousness(MD 12 min,95%CI 7.2–17.4 min)and disappearance time of symptoms(MD 6 min,95%CI–13.8–25.8 min)for patients with AAI.Larger dose of XNJ Injection(21–40 m L)may speed up the time(almost 1 h earlier).Combination of XNJ and naloxone seemed superior to the naloxone alone for al the relevant outcomes.The average difference of time in consciousness recovery was 2 h and the number of AAI patients whose consciousness recovery within 1 h was above 50%the combination group than in the control group(RR 1.42,95%CI 1.29 to 1.56).No severe adverse events or adverse reactions of XNJ were reported in the included trials.Conclusions:Low quality of evidence showed XNJ may have equal effect as naloxone and may achieve better effect as add-on intervention with naloxone for patients with AAI.We failed to evaluate the safety of XNJ Injection due to the insufficient evidence in this review.

In Silico System Pharmacology for the Potential Bioactive Ingredients Contained in Xingnaojing Injection (醒脑静注射液) and Its Material Basis for Sepsis Treatment

Objective： To elucidate the action mechanism of Xingnaojing Injection （醒脑静注射液, XNJI） for sepsis, and to target screen the potential bioactive ingredients. Methods： An integrated protocol that combines in silico target screen （molecular docking） and database mapping was employed to find the potential inhibitors from XNJI for the sepsis-related targets and to establish the compound-target （C-T） interaction network. The XNJI＇s bioactive components database was investigated and the sepsis-associated targets were comprehensively constructed; the 3D structure of adenosine receptor A2a and 5-1ipoxygenase proteins were established and evaluated with homology modeling method; system network pharmacology for sepsis treatment was studied between the bioactive ingredients and the sepsis targets using computational biology methods to distinguish inhibitors from non inhibitors for the selected sepsis-related targets and C-T network construction. Results： Multiple bioactive compounds in the XNJI were found to interact with multiple sepsis targets. The 32 bioactive ingredients were generated from XNJI in pharmacological system, and 21 potential targets were predicted to the sepsis disease; the biological activities for some potential inhibitors had been experimentally confirmed, highlighting the reliability of in silico target screen. Further integrated C-T network showed that these bioactive components together probably display synergistic action for sepsis treatment. Conclusions： The uncovered mechanism may offer a superior insight for understanding the theory of the Chinese herbal medicine for combating sepsis. Moreover, the potential inhibitors for the sepsis-related targets may provide a good source to find new lead compounds against sepsis disease.

Compatible stability study of Xing NaoJing injection based on physical-chemical properties analysis

Objective:The clinical treatment of brain diseases is urgent. Xingnaojing (XNJ) injection is often used in combination with other injection drugs. Due to the possible interaction between the injections in vivo, the particle size, osmotic pressure, pH value change and component stability decrease, that is one of the important factors causing various adverse reactions. Based on the above situation, this study investigated the physical properties and chemical composition changes of XNJ injection and its compatibility solvent and 13 kinds of clinical injection, speculated the possible interactions between the drugs in vivo from the perspective of in vitro compatibility stability, find out the safety risks of adverse reactions and provide guidance for the safe and rational use of XNJ injection. Methods:According to the clinical application, XNJ injection was mixed with 13 combination injections based on 250 mL 5% glucose injection, and placed at room temperature for 6 h. Then, the clarity, particle size, pH, osmolality, and the contents of camphor, d-borneol, and muscone of the compatible solutions were detected at 0, 1, 2, 4, and 6 h, respectively. Results:The results showed that the physical-chemical properties of compatibility solution were slightly influenced when XNJ was combined with Alprostadil injection and Danhong injection. The change of particle size and the degradation of muscone content were the main factors affecting the compatibility stability of XNJ injection, indicating that there are some problems in compatibility stability, which may be one of the causes of clinical adverse reactions. Conclusion:This study suggests that XNJ injection in combination with other injections during intravenous administration should be performed cautiously.

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

Objective： 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）. Methods： 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 punc- tually 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. Results： 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）. Conclusions： 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.