| Objective: To observe the changes of neurobehavior and cells apoptosis in the hemorragic foci and peripheral area of intracerebral hemorrhage(ICH) mice treated with bone marrow mesenchymal stem cells(BMSCs) transplantation combined with ginsenoside Rb1(GRb1), and to explore the effects of GRb1 on differentiation of exogenous BMSCs. Methods:Male C57BL/6 mice were injected with collagenase to establish the ICH models, which with neurological severity scores(NSS) higher than 8 points were used in subsequent experiments. ICH mice were randomly divided into four groups(12 mice/group): the BMSCs transplantation combined with GRb1 treatment group(BMSCs + GRb1 group), the BMSCs transplantation group(BMSCs group), the GRb1 treatment group(GRb1 group) and the model control group(MC group). On the third day post-surgery, the mouse in each group received either BMSCs transplantation combined with intraperitoneal injection of GRb1, BMSCs transplanted, intraperitoneal infection of GRb1, or no treatment. Moreover, according to the different time of sacrifice, the each group were divided into 14 d and 28 d subgroups(6 mice / subgroup). NSS was used to estimate neurological impairment of mice in each group at 1, 3, 7, 14 and 28 days after treatment. The mice brains of each group were taken and made into cryosections at the corresponding time points. The morphological changes of brain in each group were observed using hematoxylin-eosin(HE) staining. Hoechest33258 staining was used to observed the cell apoptosis surrounding the core and peripheral area of cerebral hemorrhage. Immunofluorescence staining was used to detect the expression of MAP2, GFAP and NESTIN of exogenous BMSCs. All data were analyzed statistically. Results:1. NSS: both BMSCs + GRb1 group and BMSCs group in NSS were lower than the MC group at 1, 3, 7 and 14 days(P < 0.05), GRb1 group was lower than the MC group at 1 and 3 days(P < 0.05). The BMSCs + GRb1 group decreased at 1, 3, 7 and 14 days(P < 0.05), compared with BMSCs group and GRb1 group. BMSCs group was lower than the GRb1 group at 3, 7 and 14 days(P < 0.05). 2. HE staining: each treatment group was better than the MC group in organ structure restore degree of hemorrhage area at the same time points, meanwhile the BMSCs + GRb1 group was the best one. Within each group, the organ structure restore of 28 d subgroup was better than that of 14 d subgroup. 3. Hoechest33258 staining: the numbers of cell apoptosis in both BMSCs + GRb1 group and BMSCs group were lower than the MC group at 14 and 28 days(P < 0.05). GRb1 group decreased at 14 days compared with MC group(P< 0.05), and BMSCs + GRb1 group decreased at 14 and 28 days compared with BMSCs group(P < 0.05), and BMSCs group was lower than GRb1 group at 28 days(P <0.05). 4. Immunofluorescence: 1. GFAP expression of exogenous BMSCs: GFAP-positive expressional rate of BMSCs in BMSCs + GRb1 group was higher than BMSCs group at 14 and 28 days(P < 0.05), and that of both BMSCs + GRb1 group and BMSCs group at 28 days were higher than 14 days. 2. MAP2 expression of exogenous BMSCs: MAP2-positive rate of BMSCs + GRb1 group and BMSCs group rose with the extension of time, and that of BMSCs + GRb1 group increased at 14 and 28 days compared with BMSCs group(P < 0.05). 3. NESTIN expression of exogenous BMSCs: NESTIN-positive rate of BMSCs + GRb1 group and BMSCs group declined with the extension of time, and that of BMSCs + GRb1 group was higher than BMSCs group at 14 d and 28d(P < 0.05). Conclusions: 1. In comparison to the simple treatment of either BMSCs or GRb1, BMSCs transplantation combined with ginsenodides Rb1 more can promote recovery of neurological function after intracerebral hemorrhage. 2. Both BMSCs transplantation and GRb1 treatment all were conducive to repairing tissue structure of damaged brain and suppressing cell apoptosis in the peripheral area of cerebral hemorrhage, and combined treatment would acquire better effect. 3. GRb1 could promote BMSCs differentiation into glial cells and neurons within the brain, and was advantageous to the transformation of BMSCs into neural stem cells. |
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