Intracerebral hemorrhage in rats after transplantation of neural stem cells in IL-2, IL-10 expression and the impact of Chinese intervention
|School||Central South University|
|Course||Traditional Chinese Medicine|
|Keywords||Cerebral hemorrhage Neural stem cells Transplantation immunology IL-2 IL-10 Traditional Chinese medicine|
Background ICH (Intracerebral hemorrhage, ICH) is critically ill with acute cerebrovascular disease in type, cerebral hemorrhage after the initial 30-day mortality rate was 43-51%, 75% of survivors left with varying degrees of disability, resulting in the quality of life in patients with severe decline, but also to society, the family poses a serious burden. Neural stem cells (neural stem cells, NSC) findings to the treatment of cerebrovascular disease brings new hope, but due to cerebral hemorrhage occurring endogenous neural stem cells in a limited number, not enough to repair nerve damage after cerebral hemorrhage, hence the outer endogenous neural stem cell transplantation in the treatment of cerebral hemorrhage may obtain satisfactory results. Yet brain barrier damage after cerebral hemorrhage, a large number of blood cells invade the brain, which causes the immune cell activation cytokine release. Such changes in the environment can promote brain microglial cells and other cell surface expression of major histocompatibility complex (MHC) molecules, and promote antigen presentation and recognition, neural stem cells can induce the expression of MHC molecules, the original brain \According to the immune response theory, cerebral hemorrhage BBB injury cases, treatment may occur with NSC transplantation graft-versus-host reaction (GVHD), that is mainly Th1, Th2, CTL involved in the immune response. Th1 release of many cytokines, such as IL-2, IFN-γ, etc., causing brain tissue caused by inflammation. IL-2 regardless of the inflammatory response in the brain hemorrhage or immune rejection process, plays a key role in both. Th2 secreted IL-10, IL-4 and immune closely transplantation immune tolerance. Objective: This study was designed to observe Xiehuo Bushen decoction on interleukin -2 (interleukin 2, IL-2) and interleukin -10 (interleukin 10, IL-10) in the NSC transplantation hindbrain expression of rat brain hemorrhage, explore Xiehuo kidney transplant immune response soup in NSC's role and possible mechanisms. Method: 1. Isolated and cultured for 3-5 days newborn SD rats hippocampus neural stem cells using nestin antigen immunocytochemistry, and then passaged. 2 with complement-dependent cytotoxicity assay method (CDC) in SD rats detect reactive antibody (Paner reactiveAntibody, PRA) values, select the sensitized rats were 260 high sensitivity ready for neural stem cell transplantation experiments; 3.260 rats were randomly divided into group A (control group, n = 10), B group (sham group, n = 10), C group (cerebral hemorrhage group, n = 50), D group (cerebral hemorrhage Xiehuo Bushen decoction group, n = 50), E group (cerebral hemorrhage NSC transplantation group, n = 50) and group F (cerebral hemorrhage NSC transplantation plus Xiehuo Bushen decoction group, n = 50), Ⅶ collagenase injection ICH model globus pallidus copy, intraoperative 2d for NSC transplantation after the first 2d, 4d, 7d, 14d, 28d 5 batches were killed, brain tissue samples were used RT-PCR and immunohistochemistry (SABC) was used to detect rat brain tissue IL-2, IL- 10mRNA and protein expression levels. Results 1 Immunohistochemistry (1) immunohistochemical IL-2 protein expression in the brain: normal brain tissue see a small amount of positive cells expressing each group were seen positive cells increased significantly 2d, 4d peak (P <0.05) , where E group positive cells up, 7d, 14d gradually reduced, 28d B, C, D, F group basically come down to normal, but the E group compared with the normal group remained elevated expression (P <0.01). Each group at each time point in Group E strongest expression of IL-2 (P <0.05). The same point in time than the E group F group, D group than the C group IL-2 expression was decreased (P <0.05), positive cells were mainly distributed in the hematoma surrounding tissue, according to the form of judgment primarily for oligodendrocytes. (2) Immunohistochemical detection of IL-10 protein expression in the brain: normal brain tissue see a small amount of positive cells scattered in the visible expression of each group were elevated 2d, 14d, reached the peak (P <0.05), 28d-positive cells was significantly when decreased (P <0.05). Each group at each time point in Group F strongest expression of IL-10 (P <0.05). In addition 28d outside the same time point, F group than in the E group, D group than in group C increased expression of IL-10 (P <0.05). Positively stained cells were mainly distributed in the hematoma surrounding tissue, according to the form determined mainly fibrous astrocytes glial cells. 2.RT-PCR (1) RT-PCR detection of IL-2 mRNA expression in the brain: normal group, IL-2 mRNA expression in only a very small amount, 2d when B, C, D, E, F five IL-2 mRNA expression significantly higher than group A (P <0.01), and 4d reached peak expression (P <0.05), where E is the strongest expression group (P <0.05). 7d, 14d when IL-2 mRNA expression decreased (P <0.05), 28d when the B, C, D, F 4 basic set is reduced to normal, but the E group compared with the normal group, the expression is still higher (P <0.05). The same time point, F group than in the E group, D group than the C group IL-2 mRNA expression was significantly decreased (P <0.05). (2) RT-PCR detection of IL-10 mRNA expression in the brain: normal group, a small amount of IL-10 mRNA expression. 2d after B, C, D, E, F five IL-10 mRNA expression increased compared with group A, where F group (cerebral hemorrhage neural stem cell transplantation plus Xiehuo Bushen decoction group) expressed the strongest (P <0.01), 7d gradually increased (P <0.05), 14d visible B, C, D, E four groups of IL-10mRNA peak expression (P <0.05), 28d B, C, D, E, F five group than 14d downregulated. The same time point, F group than in the E group, D group than the C group IL-10 mRNA expression increased (P <0.05). Conclusions 1. Intracerebral hemorrhage in rats after transplantation of neural stem cells upregulated IL-2, IL-10 downregulated. Xiehuo kidney transplant soup can decrease the expression of IL-2, IL-10 increases the expression. 2 Xiehuo Bushen decoction on cerebral hemorrhage after transplantation of neural stem cells of the immune rejection has tolerated effect, which may be reduced by increased IL-2 and IL-10 expression to achieve.