| Intracerebral hemorrhage is one of critical illness in the nervous system,can affect human health and quality of life. At present, the lack of specific andeffective treatment methods mainly due to the mechanism of tissue damage hasnot been fully elucidated in intracerebral hemorrhage. Relative studies clarifythat inflammation is involved in pathological process in intracerebral hemo-rrhage. In this study, human peripheral blood and perihematoma tissue inintracerebral hemorrhage were used to detect the relative data in order to clarifythe role and mechanisms of Th17cell in intracerebral Hemorrhage.This study was divided into three parts:1.48patients with intracerebral hemorrhage were selected, and detect levelsof IL-17, IL-23, IFN-γ in peripheral blood and TIM-3mRNA in PMBC in≤6h,6-24h,24-72h and>72h by ELISA and RT-PCR in order to explore the relationbetween inflammation and intracerebral hemorrhage.Level of IL-17had no significant difference in the≤6h group comparedwith the control group (P>0.05), however, level of IL-17in6-24h group,24-72h group and>72h group was significantly higher than that in controlgroup (P<0.05). Moreover, level of IL-17in24-72h group was significantlyhigher than that in≤6h group,6-24h group and>72h group (P<0.05).Level of IL-23in≤6h group,6-24h group,24-72h group and>72h groupwas significantly higher than that in control group (P<0.05). level of IL-23in6-24h group was significantly higher than that in≤6h group,24-72h group and>72h group (P<0.05).Level of IFN-γ in≤6h group,6-24h group,24-72h group and>72hgroup was significantly higher than that in control group (P<0.05).Moreover, level of IFN-γ in24-72h group was significantly higher than that in≤6h group,6-24h group and>72h group (P<0.05).Level of TIM-3mRNA in≤6h group,6-24h group,24-72h group and>72h group was significantly increased compared with control group (P<0.05).2.24patients with intracerebral hemorrhage who carry out craniotomy andhematoma removal were selected. All Specimens were divided into:≤6hgroup,6-24h group,24-72h group and>72h group according to the phasebetween onset to taking samples. Perihematoma tissue about1cm near to thehematoma was taken as experimental group and tissue far away from thehematoma was taken as control group. Pathological damage in brain tissue wasobserved by HE staining. Levels of IL-17and TIM-3were detected by IHCstaining. Levels of IL-17mRNA and TIM-3mRNA were detected by RT-PCR.Correlation analysis was done in order to clarify the role and mechanisms ofTh17cell in inflammation response and regulation mechanisms.HE staining: The pathological damages in≤6h group were almost thesame as control group, occasionally scattering inflammatory cell infiltration. In6-24h group, Perihematoma tissue was mildly injured, showing neurons andfibers edema. A number of neurons degenerated, form was incomplete. Thenumber of cells decreased slightly and nuclear pyknosis can be seen. Therewere many kinds of inflammatory cells infiltrated, mainly neutrophils.lymphocytes also can be seen. Microvascular around hematoma show conge-stion and expansion. However, in24-72h group, Perihematoma tissue was themost injured, showing neurons and fibers edema obviously. Form of manyneurons was incomplete and the number of cells decreased obviously, andnuclear pyknosis was obviously seen. There were infiltration of inflammatorycells, showing that a large number of neutrophils and lymphocytes, with a smallamount of microglia. In>72h group, the tissue damage around hematoma wasslight compared with24-72h group, the number of inflammatory cells infiltration decreased, but we still can find a few of lymphocytes, microgliashow proliferate obviously.IHC staining: level of IL-17in≤6h group in perihematoma tissue had nosignificant difference compared with control group (P>0.05), level of IL-17in6-24h group,24-72h group and>72h group in perihematoma tissue wassignificantly higher than that in control group (P<0.05). Moreover, level ofIL-17in24-72h group in perihematoma tissue was significantly higher thanthat in≤6h group,6-24h group and>72h group (P<0.05). Level of TIM-3in≤6h group in perihematoma tissue had no significant difference compared withcontrol group (P>0.05), level of TIM-3in6-24h group,24-72h group and>72h group in perihematoma tissue was significantly higher than that in controlgroup (P<0.05). level of TIM-3in24-72h group in perihematoma tissue wassignificantly higher than that in≤6h group,6-24h group and>72h group(P<0.05).RT-PCR: The expression of TIM-3mRNA and IL-17mRNA showed thesame tendency compared with IHC.Correlation analysis: level of TIM-3was positively correlated with level ofIL-17(r=0.627, P <0.01).3. Expression of NF-κBp65in perihematoma tissue was detected byWestern blot and IHC staining. Correlation analysis between IL-17andNF-κBp65was done in order to clarify the effection mechanism of IL-17/NF-κBp65pathway in perihematoma tissue.IHC staining: Expression of NF-κBp65in≤6h group,6-24h group,24-72hgroup and>72h group in perihematoma tissue was significantly higher thanthat in control group (P<0.05). Expression of NF-κBp65in24-72h group inperihematoma tissue was significantly higher than that in≤6h group,6-24hgroup and>72h group (P<0.05).Western blot: Expression of NF-κBp65neucleoprotein showed the same tendency compared with IHC.Correlation analysis: level of NF-κBp65was positively correlated withlevel of IL-17(r=0.719,P <0.01)Conclusions:(1) Level of IL-17in peripheral blood of patients with intracerebralhemorrhage increased after6h.(2) Levels of IL-23and IFN-γ in peripheral blood of patients withintracerebral hemorrhage increased.(3) Expression of TIM-3mRNA in peripheral blood monouclear cells ofpatients with intracerebral hemorrhage increased.(4) Levels of IL-17and TIM-3in perihematoma tissue increased after6h,TIM-3may aggravate the inflammation by secreation of IL-17.(5) Expression of NF-κBp65in perihematoma tissue increased afterintracerebral hemorrhage. IL-17possiblely activated NF-κB pathway whichaggravated the inflammation and damage in perihematoma tissue after intrac-erebral Hemorrhage.In this study, we were based on the human's hematoma tissue andperipheral blood in intracerebral Hemorrhage discussing mechanisms ofintracerebral hemorrhage to overcome the differences between heterogeneous.We can regulate Th17cell to degrade the inflammation and damage in perih-ematoma tissue after intracerebral hemorrhage for opening up new methods forthe treatment of intracerebral hemorrhage. |
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