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.

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.

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.

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.