| Objective: To evaluate Mechanism of NF-kappaB Signaling Pathway in Secondaryinflammatory reaction and the effects of Toll-like receptor4on the Inflammatory injury afterIntracerebral Hemmorhage(ICH) in experimental rats, and further investigate the possiblemechanism of the Secondary Inflammatory reaction induced by ICH and the Protective effectof atorvastatin.Methods: The intracerebral hemorrhage models were established by nonthrombogenicautologous blood injection method in rats. Rats were randomly divided into3groups: shamgroup; ICH group and ATV group. Each group was randomly divided into12h group,24hgroup,72h group, and7d group prespectively by different time. A needle was inserted intoright basal ganglia(coordinates:0.2mm anterior,5.5mm ventral and3.5mm lateral tobregma，6mm depth) and removed after15minutes. In contrast,50μl autologous artery bloodwas injected into the ipsilateral basal ganglia at a rate of7-10μl/min in ICH and ATV groups.The ATV groups were injected into atorvastatin (10mg kg-1d-1) by intragastricadministration. The behavioral changes were detected with neurologic deficit score by Garciamethod. By employing Brain edema changes, the water content of brain was checked up withdry/wet weight method. The histo-pathological changes were measured by Nissl staining andhematoxylin-eosin staining, The location of CD14, TLR4and around IL-1β the hematoma incerebral tissues was determined by immunofluorescence. meanwhile, the number of Positivecells and the level of protein expression about NF-κB, TLR4and IL-1β in cerebral tissues wasdetermined by immunohistochemistry and western blot.Results: Neurological function is normal on six aspects: Spontaneous activity (in cagefor5min), Symmetry of movements (four limbs), Symmetry of forelimbs (outstretchingwhile held by tail), Climbing wall of wire cage, Reaction to touch on either side of trunk,Response to vibrissae touch. Neurological function score was significantly decreased in theICH group but the score was increased in the ATV group compared with the ICH group(P<0.05). The Nissl’s body of neuron cell in ICH group were obviously lost, by contrast thenumbers of the Nissl’s body were increased in the ATV group compared with the ICHgroup(P <0.05).The result of the water content of brain: the water content of brain in cerebral tissues in ICH group was obviously increased at ICH12h, and reached the peak at72h, and was higherthan those in sham-operated group at each time point (P<0.05). Compared with the ICH group,the water content of brain of the ATV group was reduced (P<0.05).The level of the expression about NF-κB and IL-1β in the ICH group: the expression ofNF-κB and IL-1β in ICH group was significantly increased at ICH12h, reached the peak at72h, and gradually reduced at the following time. There was a correlation between theexpression of NF-κB and IL-1β(r=0.861, P<0.05); IL-1β and BWC(r=0.674, P<0.05); NF-κBand BWC(r=0.781, P<0.05).The changes of protein expression about TLR4determined by western blot in each group:Compared with the Sham group, the expression of TLR4following ICH was much higher,meanwhile the expression of NF-κB and IL-1β was markedly increased. ATV obviouslyinhibited the expression of TLR4, reduced the water content of brain, in paralleled with theimprovement of neurologic deficit score, simultaneous the expression of NF-κB and IL-1βwas lower. The results detected by Immunohistochemistry Support it.Conclusions:1. NF-κB around the hematoma may worsen inflammation and take part inthe pathological process of brain edema in the ICH rats. Its activation induced the expressionof cell factors IL-1β, and IL-1β has the tendency of increasing gradually in the ICH rats.2.TLR4can induce the inflammatory reaction by encouraging expression of NF-κB.3.Ourfindings indicate that administration of ATV has a neuroprotective effect, which may berelated to suppress inflammatory injury, thereafter, to promote the recovery of neuralfunction. |
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