| Cerebrovascular disease is one of the three diseases inducing human beings’death, and ischemic stroke which belongs to cerebrovascular disease has high mortality and disability rates, seriously threatens mankind’s health and effects their qualities of life. But, until now there haven’t been dramatic breakthroughs in the treatment of the disease, especially in the aspect of treating the patients in the recovery period of stroke.Based on our previous research, stroke belongs to collateral disease in traditional chinese medicine TCM, and its major mechanism in TCM is "the damage of toxic to cerebral collaterals". Based on the above mechanism in TCM and the latest achievements of both modern pathological physiology and bioeffects of Chinese herbs in the research of ischemic stroke, JieDuTongLuo Compound consisting of Gardenia、Salvia,、Astragalus、 Gastrodia has been developed. Therefore, this study is to research the mechanism of synaptic regeneration following cerebral ischemic injury and the beneficial effect of JieDuTongLuo Compound on this injury; to explore the effect of Chinese medicine on Cerebral ischemia inflammation levels even reaction, the improvement of brain microenvironment, reconstruction of structures and functions in synapses and the protection of neurons in order to provide a meaningful way of thinking for the acting way and targets in the treatment of ischemic cerebrovascular disease with Chinese medicine.Objective:This research demonstrate the mechanism of synapse regeneration after ischemic cerebral damage and the impact of JieDuTongLuo compound via two aspects, the structure of synapse and changes of its microenvironment.1. To verify the effect of JieDuTongLuo compound to recovery of nerve function after ischemic cerebral damage;2. To observe the changes of synapse structure after ischemic cerebral damage and the effect of JieDuTongLuo compound;3. To observe the changes of microenvironment around synapse after ischemic cerebral damage on rat and the impact of JieDuTongLuo compound.Methods:1. Prove the effect of JieDuTongLuo compound to the recovery of nerve function from ethology and morphology, including gait analysis meter, nerve function score, HE staining; 2. Observe changes of general synapse structure and its ultra microstructure via plating silver staining and transmission electron microscope TEM, and test the amount of SYP and actin via IHC;3.Observe the amount and changes of BDNF, BFGF, VEGF around microenvironment of synapse via ELISA and IHC.Results:1.To verify the effect of JieDuTongLuo compound to recovery of nerve function after ischemic cerebral damage;1) HE staining:By optical microscope we found that neurons of parietal cortex area significantly decreased4weeks and8weeks after ischemic cerebral damage on rats, and there is obvious pathological change on neurons and astroglias of cortex area, the infarct area significant decreased by effect of JieDuTongLuo compound. At high magnification lens we can see that neurons and astroglias are slightly damaged with newborn vessel, neuron and synapse.2) On the aspect of intensity of pressure in gait analysis, JieDuTongLuo compound group is significantly higher than control group.3) On the aspect of nerve function loss scale, JieDuTongLuo compound group is much lower than control group.2. Observe changes of general synapse structure and its ultra microstructure via plating silver staining and transmission electron microscope TEM, and test the amount of SYP and actin via IHC;1) Results of silver plating staining:the neuron in normal group was equably stained, had unabridged structure, many dendrite and long axon, also intensively distributed. Each synapse was tightly connected. We can see axon-dendrite synapse, axon-soma synapse, etc. However, neurons in model group was severely decreased, unevenly stained, badly pyknosis, few dendrite and hardly no axon. JieDuTongLuo compound group had more neurons, fairly equably stained, full structure, more axon and dendrite than model group, also had connect between synapse.2) Result of electron microscope:normal group:neuron had full structure, normal shape, clearly two layer of karyotheca, chromatin was evenly distributed in nucleus, organoid had normal structure. A dense synaptic with clear presynaptic membrane, postsynaptic membrane, synaptic cleft and anterior synaptic capsule. And there is also a deep staining and a large density in postsynaptic dense zone, model group:unclear structures in neuron, their membranes and nuclear membranes, chromatin was distributed mainly in the lower part of nucleus, organelles in cytoplasm were decreased with nuclear structures and mitochondria swelling. A significant reduction in the number of synaptic with nuclear presynaptic membrane and postsynaptic membrane, and a low density in postsynaptic dense zone. JieDuTongLuo compound group:a basic integrity of structure and an almost normal shape in neurons with clear nuclear membranes and well-distributed chromatin could be found. In addition, an increase in the number of some organelles such as Golgi complex, ribosome and rough endoplasmic reticulum, a slight swelling in some mitochondria could be seen. A large density in the distribution of synaptic and postsynaptic dense zone could be found. The connection between synaptic, presynaptic membrane, postsynaptic membrane and anterior synaptic capsule could also be observed clearly.3) Analysis the amount of SYN in cortex area after focal cerebral damage by IHC, we found that amount of SYN in model group was significantly lower than that in normal group after the administration for4weeksP<0.01. That of JieDuTongLuo compound group was obviously higher than model groupP<0.001. After8weeks the amount of SYN in model group is significantly lower than normal group P <0.001. SYN in JieDuTongLuo compound group showed an obvious increasing trendP<0.05, and was significantly higher than that of4weeks P<0.001. After analysing the amount of actin of cortex area after focal cerebral damage on rats via IHC, we found that after4weeks, amount of actin in model group was slightly lower than normal group. That in JieDuTongLuo compound group was higher than model group. After8weeks, the amount of actin in model group is significantly lower than normal group P<0.001. That of JieDuTongLuo compound was significantly higher than model group P<0.001, and was higher than that of4weeksP<0.05.3.Observe the amount and changes of BDNF, BFGF, VEGF around microenvironment of synapse via ELISA and IHC.The results from the content of BDNF, VEGF and BFGF both in serums and tissues of injury brains following the local brain injury in rats with the method of ELISA:1BDBF:the content of BDNF in model group decreased compared with that in control group, and showed an increasing trend compared with that in other groups without statistical significance following the administration of JieDuTongLuo compound for4weeks. After the administration of JieDuTongLuo compound for8weeks, there was an obvious decrease in the content of BDNF in model group compared with that in control groupP<0.05and an obvious increase of BDNF in JieDuTongLuo compound group compared with that in model groupP<0.05.2BFGF:the content of BFGF in model group decreased compared with that in control group, and increased compared with that in other groups with most statistical significant difference following the administration of JieDuTongLuo compound for4weeksP<0.001. After the administration of JieDuTongLuo compound for8weeks, there was an obvious decrease in the content of BFGF in model group compared with that in control groupP<0.001and an obvious increase of BFGF in JieDuTongLuo compound group compared with that in model groupP<0.001.3VEGF:the content of VEGF in model group decreased compared with that in control group, and increased compared with that in other groups with statistical significant difference following the administration of JieDuTongLuo compound for4weeks(in serum:P<0.01; in the brain homogenate:P<0.05), After the administration of JieDuTongLuo compound for8weeks, there was an obvious decrease in the content of VEGF in model group compared with that in control groupin serum:P<0.001; in the brain homogenate:P<0.05and an obvious increase of VEGF in JieDuTongLuo compound group compared with that in model groupin serum:P<0.01; in the brain homogenate:P<0.001. And these results were consistent with those from IHC.Conclusions:1. Neurons were damaged severely following the local cerebral ischemia in rat brain, and JieDuTongLuo compound had a beneficial effect on the recovery of nervous functions induced by ischemia.2. JieDuTongLuo compound could effectively promote the regeneration of neurons and synaptic after their damage mediated by local cerebral ischemia in rat brain.3JieDuTongLuo compound might promote the regeneration of synaptic via regulating the changes of some active factors in synaptic microenvironment in order to exert its effects on the protection of neurons and the recovery of nervous functions. |
PDF Full Text Request