Effects Of Xiehuo Bushen Decoction On The Expression Of Interleukin-2,Interleukin-10 In Neural Stem Cells-transplanted Immunological Rejection Of Intracerebral Hemorrhage

Objective:The study aimed to observe the effects of Xiehuo Bushen decoction on the expression of Interleukin-2,Interleukin-10 in rats brain of neural stern cells(NSCs)-transplanted experimental intracerebral hemorrhage and cultured interferon-gamma(IFN-γ)-induced cerebral microvascular endothe-lial cells(RCMECs),to explore the mechanism of Xiehuo Bushen Decoction(BSXH) in protecting RCMECs in NSCs-transplanted intracerebral hemorrhagic rat brains.Methods:1.Ninety rats were randomly divided into five groups including normal group(n=10),sham-operation group(n=20),model group(n=20),NSC-transplanted group(n=20) and BSXH-treated group(n=20).The intra-cerebral hemorrhage model was induced by injecting 0.4UⅦcollagenase into right globus pallidus(1.4mm posterior, 3.2mm lateral to bregma,and 5.6-mm depth from the cortical surface) with stereotaxic apparatus.BrdU-labled NSCs were transplanted to subventricle zone(SVZ) 2 days later.We observ-ed the expression of IL-2 and IL-10 by enzyme-linked immunosorbent assay(ELISA),western bolt and reverse transcriptase polymerase chain reaction (RT-PCR),as well as the apoptosis by TdT-mediated dUTP-biotin nick end abeling(TUNEL). 2.The cultured RCMECs were randomly divided into control group, IFN-γgroup,serum control group and XHBS serum group.The expression of IL-2 and IL-10,and apoptosis was assayed by the same technique as those in vivo respectively.Results:1.IL-2,IL-10 mRNA expressed remarkably in cultured RCMECs after induced with IFN-γ(P＜0.01 vs control group).IL-2 expression were decreased and IL-10 expression were increased after treated with BSXH (P＜0.05 vs IFN-γ,group),The cell viability of XHBS serum group were higher than that of IFN-γgroup by MTT(P＜0.05).2.The number of TUNEL positive cells which was very small in control group increased markedly in IFN-γgroup(P＜0.01 vs control group) and decreased in XH BSserum group(P＜0.05 vs serum control group).3.The expression of IL-2 mRNA and protein in RCMECs were increased remarkably after induced by IFN-γ(P＜0.05 vs control group) and decreased in BSXH serum group(P＜0.01 vs serum control group).The expression of IL-10 were gradually up-regulated in IFN-γgroup,serum control group and BSXH serum group,and there is significant difference between IFN-γgroup and BSXH serum group(P＜0.05).4.IL-2 expressed at low level in normal group and sham operation group, it was up-regulated remarkably in model group and NSC transplanted group.The expression of IL-2 was down-regulated significantly in XHBS-treated group(P＜0.05 vs NSCs-transplanted group).IL-10 mRNA also can be seen mildly in normal group and sham operation group (P＜0.05).It was higher in model group than in sham operation group (P＜0.05).The expression was up-regulated markedly in BSXH-treated group(P＜0.05 vs NSC-transplanted group).5.There was no TUNEL positive cells in the rat brains of normal group and some in sham operation group.The number of TUNEL positive cells increased significantly in model group and NSC transplanted group,and there was no difference between the two groups.The number of apoptosis cells decrea-sed markedly in XHBS-treated group(P＜0.05 vs NSC-transplanted group).6.IL-2 expressed at low level in the rat brains of normal group and sham operation group.The expression of IL-2 mRNA and protein in model group and transplanted group increased at 4d(P＜0.01 vs control group and sham operation group).It expressed mainly in neurons and endothelial cells around hematoma.it was down-regulated remarkably in XHBS-treated group(P＜0.05 vs NSC-transplanted group).There was little IL-10 mRNA expressed in normal group and sham operation group.The trend of IL-10 expression incr-eased gradually in model group, transplanted group and XHBS-treated group.The expression of IL-10 from 4d to 7d is ascendant.Conclusions:1.The expression of IL-2 and IL-10 were increased in brains of NSCstransplanted intracerebral hemorrhage rat.Xiehuo Bushen Decoction could depress IFNlammation by up-regulating the expression of IL-10and down-regulating the expression of IL-2.2.IL-10,IL-2 play a important role in brains of NSC-transplanted intrace-rebral hemorrhage rat.Xiehuo Bushen Decoction could inhibit ndothelial apoptosis by depressing the expression of IL-2 and promoting the expression of IL-10.3.Restraining the activation of IL-2 and promoting the expression of IL-10 might be the mechanism of XHBS on lessening IFNlammation to protect RCMECs.