| Objective:Establish a cerebral hemorrhage rat model induced autologous caudal artery blood injection which like the clinical symptoms of acute cerebral hemorrhage.Based on the theory of brain bowel interaction study the neuroprotective mechanisms of Liangxue Tongyu prescription treating secondary brain damage after cerebral hemorrhage from neurohistology?neuropathology and neural molecular biology.Method:1 SPF SD male rats,were divided into normal group,control group,building group.Make the cerebral hemorrhage rat model according to the method of autologous tail arterial blood injection.Observe behavior score and brain tissue morphology.2 Building modules were randomly divided into model group,Liangxue Tongyu prescription group,Tongfu Xie're prescription group,Liangxue Sanyu prescription group with 6 in each and measured after 1 day,3 days and 5 days.3 Observe and record the behavior score of rats and determine the amount of hematoma and the amount of water in the brain tissue which is to explore the influence of Liangxue Tongyu prescription in cerebral hemorrhage rat neurological function.4 By detecting rats grain number and the bowels vitality to explore the influence of Liangxue Tongyu prescription in cerebral hemorrhage rat intestinal function.5 By the immunohistochemical technique to test NGF and nNOS expression of rat brain tissue.By the nitrate reduction method to test NO content of rat brain tissue.By the method of WST-8 to test SOD content of rat brain tissue and blood.By the thiobarbituric acid method to test MDA content of rat brain tissue and blood.The above method to explore of Liangxue Tongyu prescription Interventing in intracerebral hemorrhage rats nerve repair and the brain damage reducing.6 By the immunohistochemical technique to test CCK-8 expression of rat brain and intestines tissue for observating CCK-8 after treating cerebral hemorrhage rat by Liangxue Tongyu prescription.Results:1 Cerebral hemorrhage model,building group compared with normal group,control group in ethology score significantly higher(P<0.01),and can see circular hematoma in brain tissue morphological staining.The behavioral score of contrl group was 0,without the impairment of nerve function,and the circular hematoma was not seen in brain tissue morphological staining.2 Behavioral score and brain tissue hematoma content,water content determination results2.1 Behavioral scoring:modeling of the group and the normal group,sham operation group were different degrees of neurological deficit(P<0.01).Over time,the scores of each group gradually decreased.The Logna rates of Liangxue Tongyu group were lower than the model group(P<0.05)with medicated 3days,5days.Medicated 5days Liangxue Tongyu group,Tongfu Xie're group balance beam rates lower than the model group(P<0.05).Suggesting that Liangxue Tongyu prescription and Tongfu Xie're prescription can improve the neurological deficit in rats with cerebral hemorrhage,and the effect of Liangxue Tongyu prescription is more obvious.2.2 There was no significant difference in hematoma content between the model group and the drug group after 1 day(P>0.05).The Liangxue Tongyu group,the hematoma content was lower than that of the model group(P<0.05)3 days later.5 days later Liangxue Tongyu group,Liangxue Sanyu group were lower than the model group(P<0.01,P<0.05),in which the hematoma Liangxue Tongyu more significant reduction(P<0.01).Suggesting that Liangxue Tongyu prescription,Liangxue Sanyu prescription may promote hematoma dissipated,Liangxue Tongyu prescription effect is more obvious.2.3 Determination of brain water content,modeling medicated one day each rat brain water content was significantly higher than sham group(P<0.01,P<0.05).Moisture content 3 days medicated rat model group was significantly higher than sham group(P<0.01),the brain water content of the treated groups were significantly lower than the model group(P<0.01),Liangxue Tongyu group than Liangxueshangyutang group(P<0.05).5 days after treatment,the water content of treated groups were significantly lower than the model group(P<0.01),Liangxue Tongyu group in water content below Tongfu Xie're group and Liangxue Sanyu group(P<0.05,P<0.01).Suggesting that Liangxue Tongyu prescription,Tongfu Xie're prescription,Liangxueyuyu recipe can reduce the degree of brain edema,Liangxue Tongyu prescription the most obvious effect.3 Defecation observation and intestinal function test results3.1 defecation observation,the model group,the total number of defecation,black granule were significantly less than the sham operation group(P<0.01),suggesting that decreased bowel movements after cerebral hemorrhage.Liangxue Tongyu melena group was significantly higher than the percentage of the model group(P<0.01),and melena higher percentage of grains and Liangxueshangyutang group(P<0.01).Tongfu Xie're group defecation total grains,grains and melena percentage was significantly higher than the model group(P<0.01),and significantly higher than Liangxue Tongyu group and Liangxueshangyutang group(P<0.01).Promise Liangxue Tongyu prescription,cooked rhubarb can promote defecation,improve thepost-cerebral hemorrhage constipation,which cooked rhubarb single bowel effect is more obvious.3.2 Intestinal viability assay,and the normal group at each time point sham group bowel contraction frequency,amplitude and intestinal activity were significantly higher than the model group(P<0.01),suggesting cerebral hemorrhage intestinal activity decreased.The intestinal"'viability of the model group and each group increased gradually after 3 days and 5 days.The intestinal viability of Liangxue Tongyu group was significantly higher than that of Tongfu Xie're group,and the intestinal viability of the two groups was higher than that of the model group and the cooling group(P<0.05,P<0.01).Promise Liangxue Tongyu prescription,cooked rhubarb can increase intestinal vitality,promote intestinal peristalsis.4 Restoration of brain damage and cerebral protection related indicators(NGF,nNOS,NO,SOD,MDA)detection results4.1 Expression of NGF brain tissue,the expression of NGF group Liangxue Tongyu different time than the model group,Liangxue Sanyu group(P<0.05,P<0.01),and higher than after treatment 3 days Tongfu Xie're group(P<0.05)The Suggesting that Liangxue Tongyu prescription can enhance the expression of NGF in brain tissue.4.2 nNOS expression in brain tissue,each time the content of nNOS expression model group than that in sham group(P<0.05).Liangxue Tongyu group content of nNOS expression 3 days later than the model group(P<0.05).The expression of nNOS in Tongue group,Tongfu Xie're group and Liangxue Sanyu group was lower than that in model group(P<0.01,P<0.05)after 5 days.Comparison of rat brain tissue NO content,drug 3 days model group was significantly higher than sham group(P<0.01);Liangxue Tongyu group,Tongfu Xie're group,Liangxue Sanyu group was significantly lower than the model group(P<0.01).Medicated 5 days Liangxue Tongyu group,Tongfu Xie're group was significantly lower than the model group(P<0.01,P<0.05),the lowest content of NO Liangxue Tongyu group(P<0.05).The results suggest that the drug group can reduce the expression of nNOS,reduce the NO content,Liangxue Tongyu prescription the most obvious effect.4.3 Comparison of SOD activity in rat brain,medicated 1 days,3days Liangxue Tongyu group,Tongfu Xie're of SOD activity was higher than the model group(P<0.05,P<0.01).5 days laterLiangxue Tongyu group was higher than the model group(P<0.05).Comparison of serum SOD enzyme activity,3 days Liangxue Tongyu treatment group than in model group and Liangxue Sanyu group(P<0.05,P<0.01).Suggesting that Liangxue Tongyu prescription and Tongfu Xie're prescription could enhance SOD activity and Liangxue Tongyu prescription.4.4 Comparison of MDA content in rat brain,Liangxue Tongyu treatment group 1d,3d,5d are compared with the model group(P<0.05).Tongfu Xie're group was lower than the model group after 1 day and 3 days(P<0.05,P<0.01).Liangxue Sanyu group was significantly lower than the model group after 3 days(P<0.05).Compare serum MDA content:after treatment 3 days Liangxue Tongyu group,Tongfu Xie're group were lower than the model group(P<0.01);after 5 days MDA content of the treated groups were lower than in serum of model group(P<0.05).Suggesting that the drug group can reduce the content of peroxide MDA,Liangxue Tongyu prescription the most obvious effect.5 brain tissue,intestinal tissue expression results of CCK-85.1 Expression of CCK-8 in brain tissue after medicated 1 day Tongfu Xie're group,Liangxue Sanyu group higher than sham group,model group(P<0.01,P<0.05),wherein the set of expression Tongfu Xie're highest,suggesting that Tongfu Xie're prescription can significantly promote the expression of CCK-8 protein around the hematoma 1 day after.The expression of CCK-8 was the highest in Liangxue Tongyu group after 3 days of treatment(P<0.05).The expression of CCK-8 in the 5d group was significantly higher than that in the model group(P<0.01),and the Liangxue Tongyu group was the highest(P<0.05).5.2 Expression of CCK-8 in Intestinal tissue,Liangxue Tongyu group was higher than sham group,model group,Liangxue Sanyu group(P<0.01,P<0.05)after medicated 1 day.3 day after Liangxue Tongyu group was highest and Tongfu Xie're was second,which both higher than sham group,model group,Liangxue Sanyu group(P<0.01,P<0.05).Conclusion:1 The method of cerebral hemorrhage rats autologous tail arterial blood injection is successful.2 Liangxue Tongyu prescription and Tongfu Xie're prescription both can effectively improve the rat ethology score after cerebral hemorrhage,and reduce brain damage.3 Liangxue Tongyu prescription and Tongfu Xie're prescription both can promote neurological impairment of cerebral hemorrhage rats.The effect of Liangxue Tongyu prescription is more pronounced.4 Liangxue Tongyu prescription,Tongfu Xie're prescription and Liangxue Sanyu prescription all can reduce the secondary brain damage associated with the degree of different indicators nNOS expression,reduce the neurotoxic effect of NO on brain tissue,promote SOD antioxidant effection,reduce peroxide MDA content,and then protect our brain.The effect of Liangxue Tongyu prescription is most pronounced.5 Liangxue Tongyu prescription and Tongfu Xie're prescription both can intervente rat brain tissue CCK-8 and intestinal tissue CCK-8 expression.The effect of Liangxue Tongyu prescription is more pronounced. |
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