Study On The Mechanism Of NF-κB Signaling Pathways Of The Experimental ICH Rats By The Methods Of Activating Blood And Removing Stasis And Supplementing Essence And Marrow

Purpose:The research methods of ELISA, RT-PCR, Immunohistochemistry, etc. are developed onthe hemorrhage model in rats to study whether the method of activating blood and removingstasis and supplementing essence and marrow can reduce the secondary injury mechanismafter ICH by regulating the NF-κB signaling pathways. This paper intends to demonstrate thehypothesis of whether the methods of activating blood and removing stasis and supplementingessence and marrow can reduce inflammation and apoptosis after ICH, protecting nervefunctions and promoting the repairmen of tissue damage in order to explore the mechanismfor the treatment of ICH by the methods of activating blood and removing stasis andsupplementing essence and marrow.Method:1. Normal male Wistar rats which weigh240g~260g were adopted in the experiment.They were randomly divided into sham-operated group, ICH model group (model group),hemorrhagic stroke prescription group (prescription group), and Sazengnia group (controlgroup). The author prepared stable ICH model with the method of injection in accordancewith the autologous tail arterial blood.2. Observe the intervention effect of the method of activating blood and removing stasisand supplementing essence and marrow in the neurologic impairment, cerebral water contentand pathology of brain tissue around hematoma of the rats in ICH model group by means ofneurobehavioral score, wet and dry weight method, and HE staining method.3. Detect the expression of IL-1, Il-6, and TNF-α of the brain tissue around hematoma ofICH by the application of ELISA method.4. Detect the protein expression NF-κB、TLR4、MyD88and the change of mRNA byusing immunohistochemical method and RT-PCR method.Result:1. The score of sham-operated group is0, and the prescription group and the controlgroup can obviously reduce the score of neurologic impairment compared with model group (P<0.05). The functional recovery of prescription group was the most obvious, showingsignificant differences compared with the control group (P<0.05).2. The water content of the rats in model group significantly increased compared with thesham-operated group(P<0.05); the water content of prescription group and the control groupdecreased significantly compared with model group, and prescription group decreased moreobviously, showing significant differences compared with control group (P<0.05).3. The HE staining of model group, prescription group and the control group showed thatthe pathological changes of brain tissue were obvious. Pathology in3d was more obvious thanthat in5d, but the brain damage of rats in the prescription group and control group was notserious compared with model group.4. By detecting the3d IL-1、IL-6、TNF-α content in brain tissue around hematomathough ELISA, prescription group and the control group were lower than that of model group,indicating significant differences (P<0.05), there was no significant difference of the ability ofreducing IL-1、IL-6、TNF-α between prescription group and control group (P>0.05).5. The expressions of NF-KB, TLR4and MyD88were consistent after3d cerebralhemorrhage. The expressions of positive cells were the strongest, the number was the most,and the color was the darkest, which showed brown in model group compared withsham-operated group. The expressions of positive cells were medium, the number was lessand the color was brown-yellow in control group and prescription group. The positive cellswere expressed relatively weaker in sham-operated group. The color was light yellow and thenumber of the cells was the least. There were extremely significant differences of grey valuedetection in each group compared with model group (P<0.01). The differences betweenprescription group and control group have statistical significances (P<0.05).6. The level of NF-κB mRNA、TLR4mRNA、MyD88mRNA in model groupsignificantly increased compared with the sham-operated group (P<0.01) through RT-PCR,.The level of NF-κB mRNA、TLR4mRNA、MyD88mRNA in prescription group and thecontrol group significantly decreased compared with model group (P<0.05), and there weredifferences between them (P<0.05). Conclusion:1. It is found that the method of activating blood and removing stasis and supplementingessence and marrow can significantly reduce the degree of brain edema after the cerebralhemorrhage, reduce the inflammation around hematoma which injures the brain tissue andimprove the degree of neurological impairment.2. The expressions of IL-6, IL-1and TNF-α increased in brain tissue around hematomaafter the cerebral hemorrhage. The pathological changes of the cellular form of the damagedbran tissue were closely related to them, and the method of activating blood and removingstasis and supplementing essence and marrow can restrain the expressions of IL–6, IL–1andTNF-α, fighting for the inflammation after the cerebral hemorrhage.3. The expressions of NF-κB, TLR4, MyD88increased in the damaged brain tissuearound hematoma after the cerebral hemorrhage. The mechanism of the treatment of ICH bythe method of activating blood and removing stasis and supplementing essence and marrowcan be the regulation of the signaling pathways of NF-κB-TLR4-MyD88in order toinhibit inflammatory reaction and cell apoptosis and reduce the secondary injury after thecerebral hemorrhage.