HDAC10 Alleviates Inflammation After Intracerebral Hemorrhage Via The PTPN22/NLRP3 Pathway In Rats

Background: The NLR family Pyrin domain-containing 3(NLRP3)inflammasome has a crucial role in the inflammatory process during intracerebral hemorrhage(ICH)injury,but its upstream regulatory mechanism is unclear.Protein tyrosine phosphatase,non-receptor type 22(PTPN22)is involved in cellular immune response and inflammation,and is related to a variety of autoimmune diseases.Studies have shown that PTPN22 can activate NLRP3 inflammasomes by dephosphorylating Tyr861 of NLRP3.Histone deacetylase 10(HDAC10)is a newly identified class II histone deacetylase involved in immune responses.Studies have shown that HDAC10 can directly bind to the PTPN22 promoter and repress the transcription process on PTPN22,thereby inhibiting the expression of PTPN22.However,whether HDAC10 affects the inflammatory response after ICH remains unknown.Objective: To investigate whether HDAC10 relieves ICH inflammatory injury by suppressing NLRP3 inflammasomes activation through the protein tyrosine phosphatase,nonreceptor type 22(PTPN22)pathway.Methods: ? Established a rat intracerebral hemorrhage model by injecting autologous blood into basal ganglia.Western blot was used to detect HDAC10 expression at 3h,6h,12 h,24h and 48 h after intracerebral hemorrhage in rats.?Chemically synthesized HDAC10 siRNA and PTPN22 siRNA,Western blot was used to detect the interference effect,and HDAC10 siRNA and PTPN22 siRNA with the best interference effect were screened.?Modified neurological severity score(mNSS)was used to evaluate the neurological function of rats in sham group,ICH group,ICH NC group,and ICH HDAC10 siRNA group;The dry-wet method measured brain water content in each group of rats;Evans blue(EB)detected the permeability of blood-brain barrier in each group of rats;HE staining and Nissl staining were used to detect the morphological changes of rat brain tissues;Immunofluorescence was used to detect myeloperoxidase(MPO)expression surrounding hematomas in rat brain tissues;?Western blot was used to detect the expression of HDAC10,NLRP3,cleaved-caspase-1,cleaved-IL-1 ? and cleaved-IL-18 in rat brain tissues of sham group,ICH group,ICH+NC group,and ICH+HDAC10 siRNA group;The expressions of IL-1? and IL-18 were detected by ELISA.?Western blot was used to detect of HDAC10,PTPN22,NLRP3,cleaved-caspase-1,cleaved-IL-1? and cleaved-IL-18 expression in rat brain tissues of sham group,ICH group,ICH+NC group,ICH+HDAC10 siRNA group,ICH+PTPN22 siRNA group,and ICH+HDAC10 siRNA+ PTPN22 siRNA group.Co-IP detected the binding of PTPN22 and NLRP3.Results: ? The expression of HDAC10 in the brain tissue of rats increased after intracerebral hemorrhage,reaching a peak after 24 h.?After screening,the HDAC10 siRNA(Sense: GCUGGAUUCGACUCUGCUATT;antisense : UAGCAGAGUCGAAUCCAGCTT)and the PTPN22 siRNA(Sense : GCAUGUAUGGAGUUCGAAATT;antisense : UUUCGAACUCCAUACAUGCTT)had the best interference effect.?Interfering with HDAC10 increased the neurological score,brain water content,EB extravasation and number of MPO cells,and aggravated brain tissue damage.? Interfering with HDAC10 increased the expression of NLRP3,cleaved-caspase-1,cleaved-IL-1?,and cleaved-IL-18 in rat brain tissue after intracerebral hemorrhage.?Interfering with HDAC10 increased the expression of PTPN22,thereby promoting the activation of NLRP3 inflammasomes,which in turn aggravates the inflammatory damage after intracerebral hemorrhage;and the use of PTPN22 siRNA effectively reversed this effect.In addition,HDAC10 silencing also promoted the interaction of PTPN22 and NLRP3.Conclusion:Interfering with HDAC10 can aggravate intracerebral hemorrhage injury in rats.The mechanism may be that PTPN22 induces the activation of NLRP3 inflammasomes,which aggravates intracerebral hemorrhage injury.