| 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 stem cells(NSCs)-transplanted experimental intracerebral hemorrhage,to explore the mechanism of Xiehuo Bushen Decoction (BSXH)in NSCs-transplanted intracerebral hemorrhagic rat brains.Methods:1.Rat NSCs of 3-5 day age were adopted for amplification in vitro, and detected by immunohistochemistry for the expression of Nestin as a specific mark of NSCs.2 Test the PRA of rats and Selected the high and middle sensitive of PRA rats for the following experiment;3 260 rats were randomly divided into six groups including group A (normal group)(n=10),group B(sham operation group)(n=10),group C (model group)(n=50),group D(BSXH-treated group)(n=50),group E (NSC-transplanted group)(n=50),group F(NSC-transplanted and BSXH -treated group)(n=50).The intracerebral hemorrhage model was induced by injecting 0.4UⅦcollagenase into right globus pallidus(1.4 mm posterior, 3.2mm lateral to bregma,and 5.6 mm depth from the cortical surface) with stereotaxic apparatus.while collagenase was replaced by physiologyical saline in the rats of group B and we not enforced any intervention on the rats of group A.BrdU-labled NSCs were transplanted to subventricle zone(SVZ)2 days later.We observed the expression of IL-2 and IL-10 by immunochemistry and reverse transcriptase polymerase chain reaction(RT-PCR)at the 2 nd,4 th d,7 th d,14 th d and 28 th d after the operation of NSC-transplanted.Results:1.The expression of IL-2 and IL-10 protein in brain of ratsThe protein expression of IL-2:The protein expression of IL-2 in the normal rats brain was slight.At the 2nd day after the operation,the positive cells of IL-2 was increased and arrived the peak at the 4th day(P＜0.05)in groups B,C,D,E and F.especially the group E is the highest among other groups(P＜0.01).Then they decreased gradually at 7th day and 14th day(P＜0.05)and recovered to normal level at 28th day. At the same time,the protein expression of IL-2 of group E is the highest,and expression of IL-2 mRNA in E and C group were significantly higher than F and D groups respectively(P＜0.05).The positive cells distributed at the perimeter of hematoma and subrentride zone,and most of them were digodendrocytes.The protein expression feature of IL-10:The protein expression of IL-10 in the normal rats brain was slight.At the 2nd day after the operation,the positive cells of IL-10 was increased and arrived the peak at the 14th day(P＜0.05)in groups B,C,D,E and F,and currently deceased at 28 th day(P＜0.05).and at the same point of time the group F was the highest among other groups(P＜0.01).At the same time,the protein expression of IL-10 in E and C group were significantly lower than F and D groups respectively(P＜0.05).The positive cells distributed at the perimeter of hematoma and subrentride zone,and most of them were astrocytes.2.The expression of IL-2 and IL-10 mRNA in brain of ratsThe expression of IL-2 mRNA by RT-PCR:The expression of IL-2 mRNA in the rats brain of group A was slight.At the 2nd day after the operation,the IL-2 mRNA expression of groups B,C,D,E and F occurred currently compare to group A(P＜0.01),The expression of IL-2 mRNA arrived the peak at the 4th day(P＜0.05)in group B,C,D,E and F,especially the group E was the highest among the groups(P＜0.01).then group B,C,Dand F decreased gradually at 7th day and 14 th day(P＜0.05)and recover to normal level at 28th day.but at 28 th day the expression of IL-2 was more than the expression of normal group(P＜0.05).At the same points of time except 28th day,expression of IL-2 mRNA in E and C group were significantly higher than F and D groups respectively(P＜0.05).The expression of IL-10 mRNA by RT-PCR:There was only a very slight expression of IL-10 mRNA in the rats brain of A group.the expression of IL-10 mRNA in the other groups was inceased compare to group A at the 2 nd and 7 ed day,and especially the F group was the highest among other groups(P＜0.01).arrived the peak(P＜0.05)at 14 nd day,currently decease at 28 th day(P＜0.05).At the same time except 28th day,expression of IL-10 mRNA in E and C groups were significantly lower than F and D groups respectively(P＜0.05)Conclusions:1.The expression of IL-2 are increased in brains of NSCstransplanted intracerebral hemorrhage rats.and the expression of IL-10 is decreased,Xiehuo Bushen Decoction could depress up-regulating the expression of IL-10 and down-regulating the expression of IL-2.2.IL-10,IL-2 play a important role in brains of NSC-transplanted intracerebral hemorrhage rats.Xiehuo Bushen Decoction could inhibit immune rejection response and induce immune tolerance by depressing the expression of IL-2 and promoting the expression of IL-10.
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