| The neurological damage caused by cerebral infarction has a serious impact on the health of human beings,so that the probability of disability is ranked first,and the mortality rate is third,which is lower than that of cancer and myocardial infarction.According to the WHO is expected to have about 15 million cerebrovascular disease each year,ischemic cerebrovascular disease is the incidence of 4/5.The prevalence of cerebral infarction has been gradually younger,the recurrence and disability and death rates are high,not only the quality of life of patients decreased,but also to the family and society caused a heavy blow to our medical and health.After the reduction of blood flow to the brain,the brain will produce a series of inflammatory cascade,expre ssion of inflammatory factors including amino acid toxicity,excitement,neuronal cell death,depolarization around four different mechanisms,and the mechanism in the event of a time can influence each other.The inflammatory response is the result of oxygen free radicals,cytokines and proinflammatory enzymes in the process of cascade injury.Cyt okines can be activated by the endothelial cells,the receptor activated,so that white blood cells into the brain,but also can directly stimulate microgliaor astrocytes releasing harmful ingredients,and ultimately the formation of brain damage.Combination of disease and syndrome embodies the basic theory of traditional Chinese medicine,western m edicine and the combination of the theory and experimental animal knowledge,using the theory of etiology in western medicine made of animal disease and syndrome of TCM based on the disease model,animal model with characteristics of syndromes and diseases has good stability and reliability,and the model gives the time of observation and study on the phased system so that it can be dynamic,therefore,the combination of disease and syndrome animal models has become an important development trend of modern Chinese medicine research,and has great practical significance in disease to explore the mechanism and medication on cerebral infarction.Objectives:1.Through the detection of phlegm heat in cerebral ischemia rats blood sugar index,clearing function of stress metabolism changes after cerebral ischemia,to observe the ef fect of Zha Li Nu Si recipe on regulation of metabolism disorder in cerebral infarction.2.Observe the changes of nerve function and neuron cells in rats with phlegm heat and blood stasis,proving the protective effect of Zha Li Nu Si recipe on nerve cells.3.Observe the expression of inflammatory related factors after reperfusion of focal cerebral ischemia with the phlegm heat type rats,and the effects of Zha Li Nu Si recipe on them,to clarify its possible mechanism.Methods: We separate experimental rats into six group,including untreated rats,sham operation group,pure cerebral infarction group,phlegm heat group,nimodipine group,zlnsf group;the 10 rats in the normal group,other groups according to the time points were divided into 1d,3d,7d group,n=15;except the normal group,phlegm heat model rats by other groups,complete the phlegm heat model was prepared after focal cerebral ischemia reperfusion(MCAO)model.Observation of phlegm heat in a rat model of macroscopic representation,the neurological score of rats by Longa score method(NS),hematoxylin eosin(Hematoxylin and Eeosin staining-HE)were observed in hippocampus and cortexneurons pathological changes of rat brain tissue staining method;determination of total cholesterol in model rats(total cholesterol,TC),(triglyceride,TG),(triglycerides b lood glucose,GLU),(high density lipoprotein,HDL),(low density lipoprotein,LDL)changes in energy metabolism;tumor necrosis factor by immunohistochemistry were detected inrat brain tissue tumor necrosis factor alpha(TNF-?),interleukin-6 beta(Interleukin-6?,IL-6?),intercellular adhesion molecule 1(ICAM-1),vascular cell adhesion molecule-1(VCA M-1)content,real-time fluorescence quantitative Real-time(RT-PCR)method for the determ ination of the expression of m RNA,to investigate the zlnsf effect on real brain ischemia in rat model of inflammatory cascade reaction and metabolic syndrome of phlegm heat and its neuroprotective mechanism.Results:1.Effect of zlnsf on rat nerve function of phlegm heat after cerebral ischemia reperfusion score:Sham operation group rats had no neurological function,compared with the normal group,sham operation group,the nerve function of rats after MCAO score increased significantly,compared with sham operation group,cerebral infarction group and simple phlegm heat group were 1d,3d,7d,the neurological function score were sig nificantly increased(P<0.01);especially 1d increased most significantly,and the phlegm heat model of nerve function defect than single cerebral infarction group,but the difference was not significant;after administration,effects of Zha Li Nu Si prescription at each time point of nerve function group and nimodipine group rats were decreased.As time goes on,score decreased significantly(P<0.05);Zlnsf group neurological score than nim odipine group decreased with the prolongation of time,but the difference was not signi ficant(P>0.05).2.Effect of zlnsf on phlegm heat after cerebral ischemia reperfusion in rats with blood glucose,triglyceride and cholesterol and other indicators:Compared with then ormal group,sham operation group and model group rats blood glucose and lipid content were increased to some extent,the higher blood glucose and triglyceride(P<0.05);preparation of phlegm heat model through the system,model rats blood glucose and blood lipid were increased(P<0.05);With phlegm heat group,TC,TG,HDL-C,LDL-C of ni modipine group and zlnsf groups were significantly decreased(P<0.05);GLU in 3d,7ddecreased significantly(P<0.05);The above data show that Zha Li Nu Si prescription can significantly reduce blood lipid and blood glucose level of phlegm heat in cerebral is chemia rats,and with the prolongation of time,the effect was more obvious.3.Effect of zlnsf on pathological morphology of rat brain tissue after reperfusion of phlegm heat ischemia:Zlnsf on cerebral ischemia reperfusion syndrome of phlegm heat and morphological changes of rat brain tissue:HE staining showed that in sham operation group,hippocampus structure order,cell body,clear nucleolus,uniform staining,neurons arranged closely.The model group in hippocampus cell disorder,loose structure,vacuoles were seen in the stroma of different sizes,some karyopyknosis and anachromasis,no nucleolus,neuronal cells decreased,cell shrinkage,boundary is not clear;most of the cells showed degenerative changes of nerve cells were eosinophilic.Zlnsf group and nimodipine group of nerve cells arranged disordered,still visible cell shrinkage,the peri pheral nerve cells visible mild swelling,cytoplasm,cell membrane and nucleus is able to distinguish,the degree was significantly reduced compared with model group group,zlnsf group improved significantly than nimodipine.4.Effect of zlnsf on content of inflammatory cytokines in brain tissue of rats in the syndrome of phlegm heat4.1 Effect on TNF-? expression:The effect of TNF-? in the sham operation group has a small amount of expression,pure cerebral infarction group the number of positive cells was significantly increased compared with the sham operation group(P<0.01);with the extension of time TNF-? number of positive cells decreased slowly,but still highe r than the sham operation group.With phlegm heat group,nimodipine group and zlnsf group TNF-? positive cells were decreased(P<0.01),with zlnsf 1d,3d group significantly decreased(P<0.05);Compared with the sham operation group,model group at each time point TNF-? m RNA expression of large number(P<0.01);TNF-? expression of Nimod ipine group and zlnsf group decreased remarkablely than the phlegm heat model group(P<0.01);Compared with nimodipine group,TNF-? m RNA expression of zlnsf group red uced,especially the zlnsf 1d and 7d group expression significant difference(P<0.05);Zlnsf group 3d expression had no significant difference(P>0.05).4.2 Effect on IL-6 expression:The effect of IL-6 on the expression of 1d,the numbe r of IL-6 positive cells began to increase,IL-6 reached the peak in 3d.The number of positive cells in IL-6 model group and sham operation group were significantly differe nt(P<0.05);Compared with phlegm heat model group,Zlnsf group droped obviously(P<0.05),and nimodipine group at 1d,3d was significantly different(P<0.05).These results suggest that zlnsf and nimodipine could reduce IL-6 positive cells.However,zlnsf is m ore significant.IL-6 m RNA in the sham operation group was lower,model group at each time point IL-6 m RNA expression was significantly increased(P<0.01),reached the peak at 3d;Compared with the model group,nimodipine group and zlnsf group IL-6m RNA expression decreased significantly(P<0.01);Compared with nimodipine group,zln sf group IL-6 m RNA expression decreased(P<0.05).4.3 Effect on ICAM-1 expression:The results of immunohistochemistry showed that ICAM-1 positive cells in the sham operation group was less,the expression of ICAM-1in rats with cerebral ischemia was higher than that in sham operation group(P<0.01),positive cells in one day decreased the most,and compared with the model group,the difference was significant on zlnsf group and nimodipine group rats in ischemic brain tissues(P<0.01);compared with nimodipine group,zlnsf group 1d,3d decreased significa ntly.The expression of ICAM-1 m RNA was significantly increased in the model group after cerebral ischemia reperfusion 1d,3d,7d and decreased after 3d,but was still highe r than that in sham operation group(P<0.01);Phlegm heat group in 7d when compared with the cerebral infarction group was slightly increased;the expression of zlnsf group and nimodipine group compared with the non drug group,ICAM-1 m RNA were decre ased evidently(P<0.01,P<0.05);Zlnsf group and nimodipine group ICAM-1 m RNA expression decreased significantly(P<0.05).4.4 Effect on VCAM-1 expression:In the sham operation group the expression of VCAM-1 was lower,compared with sham operation group,model group at each time poin t of the expression of VCAM-1 positive cells increased significantly(P<0.05);the expres sion was significantly decreased at each time point of the positive cells of VCAM-1 g roup(P<0.05);compared with nimodipine group,the expression of 1d was reduced signifi cantly in VCAM-1 positive group zlnsf group cell(P<0.05);In sham group,VCAM-1 m RNA has a small amount,model group at each time point VCAM-1 m RNA expression comparison sham group was sharply more(P<0.01);nimodipine group and zlnsf group VCAM-1 m RNA expression match with the model group of phlegm heat decreased re markably(P<0.01,P<0.05);Compared with nimodipine group,zlnsf prescription groups were statistically significant(P<0.05).Conclusions:1.Phlegm heat model in rats after cerebral ischemia reperfusion stress increased blood glucose,triglyceride,cholesterol,low density lipoprotein,high density lipoprotein decreased and the application of zlnsf could improve in different degree of blood glucose and blood lipid levels in rats,means that Zha Li Nu Si recipe can promote lipid metabolism after cerebral infarction.2.Phlegm heat in the rat model of nerve cells after cerebral ischemia appeared different degree of damage and showed neurological deficits,Zha Li Nu Si recipe can promote t he recovery of damaged cells and improve the nerve function defect degree.3.Expression of inflammatory factors on rats with cerebral ischemia reperfusion increased significantly,the use of Zha Li Nu Si prescription can obviously reduce the expression of inflammatory cytokines,inhibition of the inflammatory cascade reaction may be one of the neuroprotective mechanisms. |
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