| Stroke is clinical common disease,frequently occurring disease and has a high mortality rate and disability rate.Cerebral hemorrhage is extremely important types of stroke a,commonly referred to as the brain hemorrhage is a bleeding caused by rupture of non traumatic intracerebral vessels.Cerebral hemorrhage,brain edema and nerve cell apoptosis of brain tissue is an important pathophysiological process,thus preventing brain edema and apoptosis is a necessary measure for cerebral hemorrhage.Buyang Huan Wu Decoction is clinically used for the treatment of ischemic cerebral vascular diseases commonly used drugs in bleeding,can by reducing the neuronal death and promote nerve regeneration mechanism to play a role in prevention and treatment.But Buyang Huanwu Decoction play deeper mechanism of cerebral protective effect is not clear,the research field in cerebral hemorrhage and the effect of PI3K/Akt signal transduction pathway in brain tissue of rats with cerebral hemorrhage are rarely reported,the underlying mechanism is and PI3K/Akt signal transduction pathway is closely linked to the remaining questions.Therefore,research and explore the influence is positive a Buyang Huanwu Decoction on the PI3K/AKT signal transduction pathway.Preliminary studies in our laboratory found Bu Yang Huan Wu Decoction gavage administration can reduce cerebral ischemia in rat hippocampal CA1 area of damaged neurons apoptosis.In view of the cerebral ischemia and cerebral hemorrhage both in the pathogenesis and pathological damage of similar reasons.It is speculated that the inevitable can reduce brain in rat blood serum brain tissue surrounding neurons apoptosis.In this study,Rosenberg method of making rat model of intracerebral hemorrhage,Ginkgo biloba as positive control medicine,brain tissue surrounding the brain nerve function of rats were detected by blood score,moisture content and permeability of blood-brain barrier,the morphology and structure of neurons,the apoptotic cell number,cerebral hemorrhage and brain tissue pathological damage,ultrastructure of neurons in the brain the protective effect of Buyang huanwu decoction verification;immunohistochemical detection of PI3K,AKT,Bcl-2,BAX and other apoptosis related proteins and genes,to detect the changes of PI3K/AKT signal transduction pathway of Western blot protein of the two pillars of PI3K and AKT,to explore the deep mechanism of Yang also five soup effect on PI3K/AKT signal transduction pathway and its cerebral protective effect.Through these studies to ascertain the effect of Buyang Huanwu Decoction on intracerebral hemorrhage in brain tissue rats PI3K / Akt signal transduction pathway and play a role of mechanism,Buyang Huan Wu Decoction in preventing cerebral hemorrhage provide experimental basis,in order to provide new ideas and directions for the treatment of cerebral hemorrhage.Objective:Explore Buyang Huanwu Decoction on cerebral hemorrhage rat brain protective effect,the effect of apoptosis related genes and proteins,and PI3K/Akt signal transduction pathway,explore the mechanism of its protective effect on the brain.Method:1 Model preparation and grouping:Clean grade SD healthy male rats were randomly divided into sham operation group,model group(ICH group),ginkgo leaf group(positive control group),Buyang Huanwu Decoction in the low,medium and high dose group.The ginkgo leaf,Buyang Huanwu Decoction in the low,medium and high dose group gavage administration,sham operation group,model group given the same amount of water.SD rats weighing,according to the Rosenberg method to make the model,select the successful model rats to test the outcome of the relevant indicators of drug intervention.2 Neurological function score:After surgery to conscious SD rats appeared ipsilateral Horner syndrome: no direction of autonomous motion,provided at the end of the rotated to the right side,by circling the right and right forelimb occurred paralysis or right forefoot adduction,wrist will buckling,bending head toward the right side of the back.These symptoms can be used as a model of success criteria.3 The detection of brain water content:The analysis of balance of brain tissue after brain tissue was weighed,placed in the temperature of 160 DEG in the oven dried to constant weight.The formula for calculating the water content of brain tissue: brain water content(%)=(wet weight reduce dry weight / wet weight)* 100%.4 Blood brain barrier permeability test:The changes of permeability of blood brain barrier were detected by EB tracer method and electron microscope.5Brain tissue pathological damage detection:HE staining and electron microscope were used to observe the pathological changes of brain tissue and the changes of neuronal in SD rats after cerebral hemorrhage.6 The number of apoptotic cells:To detect the change of apoptotic cell number in situ by using the method of nick end labeling(TUNEL).7 PI3K,AKT,Bcl-2,BAX and other protein expression detection:The expression changes of PI3K,AKT,Bcl-2,BAX and other proteins were detected by IHC method,and the changes of AKT and PI3K were detected by blot Western method.Result:Buyang Huanwu Decoction gavage after administration,can reduce the number of neuronal apoptosis in rats with intracerebral hemorrhage,reduce the blood brain barrier(BBB),reduced brain water content,maintenance of the neuronal morphology and structure stability,effectively reduce the neurological assessment,can realize the neuroprotective effect of cerebral hemorrhage.By regulating the apoptosis related proteins and gene changes,activation of PI3K/AKT signal transduction pathway to achieve the role of.1.The effect of SD on neurological function score:Rats in the model group,Yang also five soup low dose group rats neural function has not been improved significantly(P>0.05)and Buyang Huanwu Decoction,high dose group,ginkgo leaf group rats neural function was significantly improved(P <0.05,P<0.01).2 The effect of brain tissue ultrastructure in SD rats:Model group,neurons were partially dissolved,nuclear membrane is continuous,the nuclear leakage,chromatin condensation,empty,cytoplasmic most of the missing,the remnants of dense mitochondria,mitochondrial cristae is not easy to distinguish and other organelles decreased significantly.Buyang Huanwu Decoction in low,middle,high dose group and ginkgo leaf in the treatment group after neuronal injury was relieved,clear karyotheca,chromatin distributed evenly,rough endoplasmic reticulum and ribosome rich,mitochondria,although some mitochondria cristae,but most have been close to normal,the number of apoptotic cells decreased significantly.3 The effect of SD on brain tissue water content in rats:Model group of rat brain tissue water content was significantly higher than that in the sham operation group(P<0.05),Ginkgo biloba,Buyang Huanwu Decoction in the low,middle and high dose group was significantly lower than that of model group(P<0.05);Buyang Huanwu Decoction,high dose group and ginkgo leaf group differences had no statistical significance(P >0.05,P<0.01).4 The effect of SD on the permeability of blood brain barrier in rats:Model group,the content of Evans blue was significantly higher than that in sham operation group(P<0.05),Ginkgo biloba,Buyang Huanwu Decoction in the low,middle and high dose group was significantly lower than that of model group(P<0.05,P<0.01);Buyang Huanwu Decoction,high dose group and ginkgo leaf group differences had no statistical significance(P>0.05).5 The effect of brain tissue pathological injury in SD rats:He staining and electron microscope were used to observe the ultrastructure of neurons in the,in maintaining the structural stability of neurons and inhibition of apoptosis,Buyang Huanwu Decoction low,high dose group and ginkgo leaf group showed better therapeutic effect and Buyang Huanwu Decoction high dose group and ginkgo leaf group is more prominent.6 The effect of SD on cell apoptosis in rats:The number of apoptosis cells of model group was significantly higher than that in sham operation group(P<0.05),ginkgo leaf,Buyang Huanwu Decoction in the low,middle and high dose group was significantly lower than that of model group(P<0.05,P<0.01);Buyang Huanwu Decoction,high dose group and ginkgo leaf group differences had no statistical significance(P>0.05).7 The effect of PI3K,AKT,Bcl-2,BAX protein expression in brain tissue of SD rats:PI3K,Akt,bcl-2: the number of positive expression of protein in model group was higher than those in the sham operation group(P<0.05),Ginkgo biloba leaves,Buyang Huanwu Decoction in the low,middle and high dose group was significantly higher than that of model group(P<0.05,P<0.01);Buyang Huanwu Decoction,high dose group and ginkgo leaf group differences had no statistical significance(P >0.05).Bax: number of positive expression of protein in model group was significantly higher than that in the sham operation group(P<0.05),ginkgo leaf,Buyanghuanwu decoction also five soup of low,middle and high dose group was significantly lower than that of model group(P<0.05,P<0.01);Buyang Huanwu Decoction,high dose group and ginkgo leaf group differences with no statistical significance(P>0.05).Conclusion:Ultimately Buyang Huanwu Decotion of cerebral hemorrhage caused by brain damage resistance mechanism to activate the PI3K/AKT signal pathway,regulation of the Bcl-2/Bax ratio,inhibit neuronal apoptosis,reduced BBB permeability,reduce brain edema. |
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