The Influence Of Function And Mechanism Of Drugs For Inducing Resuscitation On Blood-brain Barrier And Brain Cells

Objective:To explore the influence mechanism of drugs for inducing resuscitation on BBB and brain cells function. From the whole-target site-cellular and molecular level to explain its opening Cardiac orifice and awakening consciousness "belong to Heart Sutra"(the heart storing spirit, heart governing mind)and theory of drug property and scientific connotation,and to provide reference to cliniCal therapy.Methods:Using the method of combining integral animal experiments and cell culture, using PC 12 cells and mouse brain microvascular endothelial cells (bEnd.3) as an breakthrough point,combined with spectrophotometry and test methods of molecular biology to explore influence mechanism of drugs for inducing resuscitati on on brain function.①using mouse bilateral Carotid artery ligation was investigated drugs for inducing resuscitati on acute cerebral ischemia in mice brain protection effects.②using Evans blue,and determination of the effects in blood-brain barrier permeability that drugs for inducing resuscitati on acute cerebral ischemia in mice.③By MTT method and spectrophotometry to determine of activity of direct dosing and mediCated serum to Hypoxia-ischemia PC12 and Bend.3 cell,while in accordance with the determination of kit required to measure Ca2+ content,NO,Na+-K+-ATPase content.④To determine the effect about drugs for inducing resuscitation of PC 12 cells PLCβ1mRNA expression by RT-PCR.Result:①Survival time of mice in Borneol group signifiCantly longer (P<0.05), Musk group although trend to extend the survival time of mice, but no significant difference (P> 0.05); Musk,Borneol and Benzoin group compared with the control group,SOD was signifiCantly increased(P<0.05),Musk,Borneol,Benzoin and Storax group,MDA decreased significantly(P<0.01).②Compared with model group,EB conten of the Musk,Borneoland Benzoin group were decreased, and EB in Borneo group were signifiCantly decreased(P<0.05);③PC12:compared with model group,MTT values in Musk 2.5μg/100μl,Borneoll,1.25,2.5μg/100μl,Benzoin18.75,37.5,56.25μg/100μl,Storax 12.5,25,37.5μg/100μl group were significantly higher (P<0.05). Compared with model group,the cell Ca2+ concentration in Borneo 1,1.25,2.5μg/100μl groups,Storax 12.5μg/100μl group were significantly lower than the model group(P<0.05). Compared with model group, Musk,Borneol,Benzoin and Storax each dose group,NO levels were significantly higher (P<0.05). Compared with the model group,MTT values in Musk33.33,50,66.67mmg/kg,Borneol100,200mmg/kg,Benzoin 1000mmg/kg groups were significantly higher (P<0.05). Compared with control group and tween group,Ca2+ in Musk,Borneol,Benzoin and Storax each dose groups were signifiCantly higher (P<0.05),but with the model group,the medicated serum groups Ca2+ content ratio were significantly lower than model group (P<0.05). compared with the model group,the cell Na+-K+-ATPase of Musk66.67mg/kg group,Borneol 200 mg/kg group,Storax 666 mg/kg group were significantly increased (P<0.05). Compared with model group,musk 66.67mg/kg Group PLC-β1mRNA content showed high expression (P<0.05).⑤bEnd.3:Musk,Borneol,Benzoin and Storax each dose group compared with the solvent group,MTT value were higher than the solvent group (P<0.05, P<0.01). The intracellular Ca2+ content of Musk 2.5μg/100μl,3.75μg/100μl group,Borneol groups,Benzoin 37.5μg/100μl,Styrax 25μg/100μl group was significantly lower (P <0.05). Musk,Borneol,Benzoin and Storax each dose group MTT values increased significantly (P<0.05). Compared with blank group and tween group,Ca2+ in Musk 33.33mg/kg Benzoin 1000mmg/kg groups were significantly decreased(P<0.05). Compared with blank group,Musk 33.33mg/kg group,Borneol 150 mg/kg group,Benzoin 1000 mg/kg group,Storax 666 mg/kg group cell Na+-K+-ATPase activity was significantly decreased (P<0.05).Conclusion:Drugs for inducing resuscitation for hypoxic brain injury with the exact protective effect:①Borneol is the key drugs of cerebral protection,which can extend the survival time of mice, reduce hypoxia-ischemia animal blood-brain barrier permeability, stabilize BBB structure, it's stronger. the role of strength of Musk,Benzoin and Storax that protect against hypoxic-ischemic brain is similar.②Drugs for inducing resuscitation can increase brain SOD activity and lower MDA content, to control brain cell damage.③drugs for inducing resuscitation and its metabolites on hypoxic injury in PC12 cells also have a signifiCant protective effect.On the physiological state Bend.3 cells have some growth-promoting effect.The mechanism may be,closely related:①significantly increased the damaged cell NO release and reduce the excessive Ca2+ influx; increased Na+-K+-ATPase content,to promote Na+/Ca2+ exchange,accelerate cell Ca2+ efflux; inhibition PLC-β1 hydrolysis,reducing the concentration of intracellular Ca2+, intracellular Ca2+ co-inhibition of overloading,the maintenance of intracellular Calcium homeostasis. It is a key factor of cerebral protection.②Can inhibit Na+-K+-ATPase content,which inhibits ATP-dependent drug efflux pumps,thus play a role in brain.Drugs for inducing resuscitation against cerebral ischemia and hypoxia about mice in physiological and pathological conditions, reduce the blood-brain barrier permeability in hypoxic-ischemic state to play a role in brain protection, reduce the death rate of infection of endotoxin, promote the physiological state of blood-brain barrier permeability in mice significantly. "Heart Storing Spirit" "Brain is the house of the original spirit" "Heart and brain forged", "acrid-opening" is the key of it play Open-minded manner, awakening consciousness, to explan "Xin Wen-hearts by-floating trends-enlighten Stasis,fails to grasp the effect of aromatic medicinal properties of the system characteristics.