Effects Of Extract Of Leave Ginkgo Biloba On Ultrastructure Of Apoptotic Neurons That Surround Intracerebral Hemorrhage In Rats

Objective To study the protection of extract of leave Ginkgo Biloba on neurons that surround intracerebral hemorrhage(ICH)in rats.Method ICH model was established by injecting collagenaseâ…¦into the brain of rats. 144 male rats were randomly divided into the extract of leave Ginkgo Biloba remedial group and the contrast groups including ICH contrast group and normal contrast group without ICH. Then the extract of leave Ginkgo Biloba remedial group were divided into 4 groups which were extract of leave Ginkgo Biloba remedial group of 24 h before the operation and extract of leave Ginkgo Biloba remedial group of 0 h, 6 h and 24 h after the operation. All rats involved in the experiment were killed on the 3, 7 d after ICH. Method of Narrow-alley Test were applied to study behavioral index. Immunohistochemical staining technique was used to demonstrate expressions of Bcl-2, Bax and caspase-3 protein. The ultrastructure of neurons that surround intracerebral hemorrhage in rats was observed by transmission electron microscope and the mitochondria and nerve synapse was performed a stereology analysis.Results The result of neural behavioral test,After recovery from anesthesia, Rats in each group suffered from symptoms of hemiplegia, Manifested as cerebral hemorrhage contralateral limbs to reduce or limb weakness, forelimb is the most serious. The neurological deficit in the majority of rats showed crawling or can not stand rotating, active motion decrease, and so on. With the passage of time, the treated rats gradually reduce the neurological deficit. The result of immuno- histochemistry,the positive expression of Bcl-2 in treatment group is higher than in model group (P<0.01);However the positive expression of Bax and Caspase-3 in treatment group degraded prominently. The protein ratio of Bcl-2/Bax increased. Comparing with extract of leave Ginkgo Biloba remedial group of 24 h before the operation and extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation, the positive expression of Bcl-2 in 24 h treatment group after the operation decreased,the positive expression of Bax and Caspase-3 increased,the reesults had prominently difference (P<0.05). Comparing with extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation, the positive expression of Bcl-2 in extract of leave Ginkgo Biloba remedial group of 24 h before the operation increased, the positive expression of Bax and Caspase-3 decreased,the reesults had prominently difference (P<0.05). The positive expression of Bcl-2, Bax and Caspase-3 in 0 h treatment group and 6 h treatment group had no prominently difference (P>0.05). The ultrastructure of neuron,In normal group, ultrastructure of neurons was normal. In model group, neurons in animals showed the presence of shrinked and depressed nuclei, peripherally located chromatin, pycnosised karyopycnosis nucleus, fewer nucleoli and broden rough endoplasmic reticulum. In addition, neuronal mitochondria in this group demonstrated an increased volume, collapsed crista and cavity-like degeneration, and in most cases, were swelled. Structure of synapse membranes clouding, synaptic vesicles decreasing. The number of nerve synapse decreased prominently. Meanwhile, the clearance of synapse widen;The compact area behind synapse thinningz. In extract of leave Ginkgo Biloba-treated group, the neuronal mitochondria and rough endoplasmic reticulum appeared more normal than those of model group, and the degenerating ultastructure was more improved leave Ginkgo Biloba remedial group of 24 h after the operation than extract of leave Ginkgo Biloba remedial group of 24 h before the operation and extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation. Stereology analysis of nerve mitochondria, Compared with model group,volume density, numerical density, specific surface and specific membrane of the extract of leave Ginkgo Biloba treatment group increased notably (P<0.01). Compared with extract of leave Ginkgo Biloba remedial group of 24 h before the operation and extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation, volume density,numerical density, specific surface and specific membrane of the 24 h treatment group decreased (P<0.05). Comparing with extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation, the parameters of stereology of nerve mitochondria in leave Ginkgo Biloba remedial group of 24 h before the operation increased,the reesults had prominently difference (P<0.05). Stereology analysis of nerve synapse displayed that the number of nerve synapse increased prominently in treatment group than in model group (P<0.01);Meanwhile, the clearance of synapse becomed narrow conspicuously (P<0.01);The compact area behind synapse thicken obviously (P<0.01). The visual parameters of nerve synaptosome in 24 h treatment group after surgery were different compared with extract of leave Ginkgo Biloba remedial group of 24 h before the operation and extract of leave Ginkgo Biloba remedial group of 0 h, 6 h after the operation.Conclusionâ‘ Extract of leave Ginkgo Biloba can protect neuron around hemorrhagic focus in rat brain and reduce neuron apoptosis.â‘¡Extract of leave Ginkgo Biloba may be a potential drug for ICH patients for its possible effect of inhibiting apoptosis that depents on mitochondrial dysfunction.â‘¢Extract of leave Ginkgo Biloba may promote the reconstruction and functional enhancement of synaptic ultrastructures, facilitating the reconstruction of neural network and increasing brain plasticity.â‘£The effect was better if it was used 24 h before the operation.