Ｉｎ ｏｒｄｅｒ ｔｏ ｒａｉｓｅ ｔｈｅ ｅｆｆｉｃｉｅｎｃｙ ｏｆ ｃｈｅｍｏｔｈｅｒａｐｙ ｏｎ ｍａｌｉｇｎａｎｔ ｔｕｍｏｒ，ｗｅ ｔｒｅａｔｅｄ ｔｒａｎｓｐｌａｎｔｅｄ ｈｅｐａｔｏｃａｒｃｉｎｏｍａｂｅａｒｉｎｇ ｍｉｃｅ ｗｉｔｈ Ｇｅ１３２（ｏｒｇａｎｉｃ ｇｅｒｍａｎｉｕｍ）．Ｔｈｅ ｒｅｓｕｌｔｓ ｗｅｒｅ∶① ｉｎ ｔｈｅ ｇｒｏｕｐ ｏｆ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２，ｔｈｅ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｗａｓ ３１０％，＋＋＋ ｗｈｉｔｅ ｂｌｏｏｄ ｃｅｌｌ（ＷＢＣ）ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｒａｎｓｐｌａｎｔｅｄ ｔｕｍｏｒ ｗａｓ ７２７％，ａｎｄ ｔｈｅ ｓｅｒｕｍ ｓｕｐｅｒｏｘｉｄｅ ｄｉｓｍｕｔａｓｅ （ＳＯＤ） ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ １２１ Ｎｕ／ｍｌ； ② ｉｎ ｔｈｅ ｃｏｎｔｒｏｌ ｇｒｏｕｐ ｗｉｔｈ ｎｏｒｍａｌ ｓａｌｉｎｅ ｓｏｌｕｔｉｏｎ （ＮＳ），＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｕｍｏｒ ｗａｓ ３６．４％， ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ ８１ Ｎｕ／ｍｌ，ａｎｄ ｔｈｅｒｅ ｗａｓ ｓｉｇｎｉｆｉｃａｎｔ ｄｉｆｆｅｒｅｎｃｅ （Ｐ＜０．０５） ｂｅｔｗｅｅｎ ｔｈｅ ｔｗｏ ｇｒｏｕｐｓ；③ ｉｎ ｇｒｏｕｐ ｏｆ ｃｙｃｌｏｐｈｏｓｐｈａｍｉｄｅ ａｄｍｉｎｉｓｔｒａｔｉｏｎ，ｔｈｅ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｗａｓ ３７．０％，＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｈｅ ｔｕｍｏｒ ｗａｓ ２７．３％，ａｎｄ ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｒａｔｅ ｗａｓ １０２ Ｎｕ／ｍｌ；ａｎｄ ④ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２ ｉｎ ｃｏｍｂｉｎａｔｉｏｎ ｗｉｔｈ ｃｙｃｌｏｐｈｏｓｐｈａｍｉｄｅ，ｓｈｏｗｅｄ ａ ｃａｒｃｉｎｏｓｔａｔｉｃ ｒａｔｅ ｏｆ ４５．０％，＋＋＋ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ｆｏｕｎｄ ｉｎ ｔｕｍｏｒ ｗａｓ ５４．５％，ｔｈｅ ｓｅｒｕｍ ＳＯＤ ａｃｔｉｖｉｔｙ ｗａｓ １４２ Ｎｕ／ｍｌ．Ｉｎ ｃｏｍｐａｒｉｓｏｎ ｂｅｔｗｅｅｎ ｇｒｏｕｐ ２ ａｎｄ ５，ｔｈｅｒｅ ｗｅｒｅ ｓｉｇｎｉｆｉｃａｎｔ ｄｉｆｆｅｒａｎｃｅ （Ｐ＜０．０５） ｉｎ ｂｏｔｈ ｔｉｓｓｕｅ ＷＢＣ ｉｎｆｉｌｔｒａｔｉｏｎ ａｎｄ ＳＯＤ ａｃｔｉｖｉｔｙ．Ｔｈｅｓｅ ｒｅｓｕｌｔｓ ｓｕｇｇｅｓｔ ｔｈａｔ ｈｉｇｈｄｏｓａｇｅ Ｇｅ１３２ ｔｒｅａｔｍｅｎｔ ｃａｎ ｅｎｈａｎｃｅ ｔｈｅ ｃｅｌｌ ｍｅｄｉａｔｅｄ ｉｍｍｕｎｉｔｙ ａｎｄ ａｃｔｉｖｉｔｙ ｏｆ ｓｅｒｕｍ ＳＯＤ ｏｆ ｔｒａｎｓｐｌａｎｔｅｄ ｈｅｐａｔｏｃａｒｃｉｎｏｍａｂｅａｒｉｎｇ ｍｉｃｅ ｗｉｔｈ ｃｅｒｔａｉｎ ａｎｔｉｃａｒｃｉｎｏｍａ ｅｆｆｅｃｔ．
Objective To study the transfection and expression of pleiotrophin (Ptn) gene in mice adipose-derived stem cells (ADSCs) so as to provide a new approach for the treatment of ischemic injury. Methods ADSCs from clean inbred C57BL/6W mice (weighing, 15-20 g) were isolated and cultured in vitro. The cell surface markers (CD29 and CD44) of ADSCs were identified by flow cytometry. The ADSCs were transfected with plasmid pIRES2-LEGFPN1 (containing Ptn gene coding sequence) as experimental group (group A) and with plasmid pLEGFP-N1 (containing GFP gene coding sequence) as control group (group B). After ADSCs were transfected by different plasmids respectively, the cells containing Ptn gene were selected by G418 (the best selected concentration was 200 μg/mL), and the immunophenotype of the cells was identified by flow cytometry after transfection. Meanwhile, real-time fluorescence quantitative PCR and Western blot were used to analyse the expression levels of Ptn mRNA and PTN protein in selected cells. Results The mice ADSCs were isolated and cultured successfully in vitro. The positive rates of the cell surface markers CD29 and CD44 of ADSCs were 99.5% and 95.8%, respectively; the double positive rate of CD44 and CD29 was 93.6%. The positive rates of the cell surface markers CD29 and CD44 of ADSCs were 99.1% and 95.6%, respectively after transfection of Ptn gene; the double positive rate of CD44 and CD29 was 93.4%. The expression levels of Ptn gene and PTN protein in group A were significantly higher than those in group B (P lt; 0.05). Conclusion The ADSCs can be stablely transfected by Ptn gene, the transfected ADSCs can express PTN protein highly, which is a new idea for tissue engineering of vascular reconstruction.
Objective To establish a stable colorectal cancer model in liver specific insulin-like growth factor (IGF)-1 deficient (LID) mice and examine the potential relationship between IGF-1 level and risk of mice constitutional colorectal cancer. Methods ①Establishment of a colorectal cancer model: The LID mice, in which IGF-1 level in circulation was 25% of BALB/c mice. Induction of colorectal cancer was achieved by using the 1,1 Dimethylhydrazine (DMH) with hypodermic injection at transverse part. ②Eighty fresh samples of cancer tissues and adjacent tissues were obtained from LID mice (experimental group) and BALB/c mice (control group). The expression of IGF-1 was studied by immunohistochemical assay (SP method). Results ①Weight loss occurred in both experimental group and control group after injection. Compared with the body weight before injection on 18 weeks and 24 weeks in each group, there were significant differences after injection at the same phase in each group (P＜0.05). ②The results of IGF-1 expression in cancer tissues and adjacent tissues: IGF-1 got a diffuse distribution in cancer cell cytoplasm. The positive expressions of IGF-1 in the cancer tissues and their adjacent cancer tissues were 6/7, 2/7 and 13/16, 7/16 respectively in experimental group and control group. There were significant differences between the cancer tissues and adjacent tissues inside both groups (P＜0.05). There were no significant differences inside both of cancer tissues and adjacent tissues respectively between experimental group and control group (Pgt;0.05). Conclusion In the established colorectal cancer model by DMH, IGF-1 plays an important role in the development and progression of colorectal cancer.
Objective To observe the expressions of CXC chemokine receptor 4 (CXCR4) in muscle satell ite cells in situ of normal and cardiotoxin-intoxicated muscle tissues so as to further investigate the molecular mechanism involving inmuscle regeneration such as progressing muscular dystrophy (PMD) for seeking the way to cure muscle retrogression. Methods The muscle injured model of 12 C57 male mice was made by injecting cardiotoxin (5 μg per mouse) in left quadriceps femoris, their right quadriceps femoris was used as control without any injection. The histological, immunohistochemical analysis and RT-PCR were done to investigate the expression of CXCR4 in the quadriceps femoris in situ after 1 day, 4 days, 1 week, 2 weeks, 4 weeks and 6 weeks. Results HE staining results demonstrated that the muscle tissues experienced the process from muscle injury, repair to regeneration. The result of immunohistochemistry showed that the expressions of CXCR4 in injured muscle tissue were 1 955.6 ± 150.3, 2 223.2 ± 264.3, 2 317.6 ± 178.7, 3 066.5 ± 269.6, 1 770.9 ± 98.7 and 1 505.7 ± 107.1 at 1 day, 4 days, 1 week, 2 weeks, 4 weeks and 6 weeks after injection of cardiotoxin, there was significant difference when compared with normal muscle (640.3 ± 124.0, P lt; 0.001). The RT-PCR showed that the expressions of CXCR4 mRNA in injured muscle tissue were0.822 ± 0.013, 0.882 ± 0.025, 1.025 ± 0.028, 1.065 ± 0.041, 0.837 ± 0.011 and 0.777 ± 0.015 at 1 day, 4 days, 1 week, 2 weeks, 4 weeks and 6 weeks after injection of cardiotoxin, there was significant difference when compared with normal muscle (0.349 ± 0.006, P lt; 0.001). Conclusion CXCR4 may be the critical protein in the process of muscle impairment and reparation.
ObjectiveTo study the effect and mechanism of lipopolysaccharide (LPS) on osteoclasts formation and its bone resorption function.MethodsBone marrow-derived macrophages (BMMs) were extracted from the marrow of femur and tibia of 4-week-old male C57BL/6 mice. Flow cytometry was used to detect BMMs. The effect of different concentrations of LPS (0, 100, 200, 500, 1 000, 2 000 ng/mL) on BMMs activity was examined by cell counting kit 8 (CCK-8) activity test. In order to investigate the effect of LPS on osteoclastogenesis, BMMs were divided into macrophage colony-stimulating factor (M-CSF) group, M-CSF+receptor activator of nuclear factor κB ligand (RANKL) group, M-CSF+RANKL+50 ng/mL LPS group, M-CSF+RANKL+100 ng/mL LPS group. After the completion of culture, tartrate resistant acid phosphatase (TRAP) staining was used to observe the formation of osteoclasts. In order to investigate the effect of LPS on the expression of Connexin43, BMMs were divided into the control group (M-CSF+RANKL) and the LPS group (M-CSF+RANKL+100 ng/mL LPS); and the control group (M-CSF+RANKL), 50 ng/mL LPS group (M-CSF+RANKL+50 ng/mL LPS), and 100 ng/mL LPS group (M-CSF+RANKL+100 ng/mL LPS). The expressions of Connexin43 mRNA and protein were detected by Western blot and real-time fluorescent quantitative PCR, respectively. In order to investigate the effect of LPS on osteoclast bone resorption, BMMs were divided into M-CSF group, M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group. Bone absorption test was used to detect the ratio of bone resorption area.ResultsThe flow cytometry test confirmed that the cultured cells were BMMs, and CCK-8 activity test proved that the 100 ng/mL LPS could promote the proliferation of BMMs, showing significant differences when compared with the 0, 200, 500, 1 000, and 2 000 ng/mL LPS (P<0.05). TRAP staining showed no osteoclast formation in M-CSF group. Compared with M-CSF+RANKL group, the osteoclasts in M-CSF+RANKL+50 ng/mL LPS group and M-CSF+RANKL+100 ng/mL LPS group were larger with more nuclei, while the osteoclasts in M-CSF+RANKL+100 ng/mL LPS group were more obvious, and the differences in the ratio of osteoclast area between groups were statistically significant (P<0.05). Western blot result showed that the relative expression of Connexin43 protein in LPS group was significantly higher than that in control group (P<0.05). Real-time fluorescent quantitative PCR showed that the relative expression of Connexin43 mRNA in control group, 50 ng/mL LPS group, and 100 ng/mL LPS group increased gradually, and the differences between groups were statistically significant (P<0.05). Bone resorption test showed that osteoclast bone resorption did not form in M-CSF group, but the ratio of bone resorption area increased gradually in M-CSF+RANKL group, M-CSF+RANKL+50 ng/mL LPS group, and M-CSF+RANKL+100 ng/mL LPS group, and the differences between groups were statistically significant (P<0.05).ConclusionLPS at concentration of 100 ng/mL can promote the expression of Connexin43, resulting in increased osteoclastogenesis and enhanced osteoclastic bone resorption.
Objective To investigate the possibility of gene therapy of osteolysis around artificial joint prosthesis by constructing the recombinant adenovirus which can silence tumor necrosis factor α (TNF-α). Methods The primer of small interfering RNA (siRNA) coding sequence of silent TNF-α was designed and amplified, and then RAPAD adenovirus packaging system was used to load the sequence to adenovirus, and the recombinant adenovirus Ad5-TNF-α-siRNA-CMVeGFP which lacked both E1 and E3 regions was constructed. Then 64 female BABL/C mice (weighing, 20-25 g) were randomly divided into 4 groups (n=16): blank control (group A), positive control (group B), simple adenovirus (group C), and treatment group (group D). The prosthetic-model was established in group A, and the prosthetic-loosening-model in groups B, C, and D. At 2 weeks after modeling, PBS solution was injected first, and then the same solution was injected 24 hours later in group A; titanium particle solution was injected, and then PBS solution, Ad5 E1-CMVeGFP (1 × 109 PFU/mL), and Ad5-TNF-α-siRNA-CMVeGFP (1 × 109 PFU/mL) were injected, respectively in groups B, C, and D 24 hours later, every 2 weeks over a 10-week period. The general condition of mice was observed after operation. The tissues were harvested for histological observation, and the expression of TNF-α was detected by Western blot at 12 weeks after operation. Results The positive clones were achieved by enzyme digestion and confirmed by DNA sequencing after loading the target genes into adenovirus vector, and then HEK293 cells were successfully transfected by recombinant adenovirus Ad5-TNF-α-siRNA-CMVeGFP. All mice survived to the completion of the experiment. Histological observation showed that there were few inflammatory cells and osteoclasts in group A, with a good bone formation; there were a large number of inflammatory cells and osteoclasts in groups B and C, with obvious bone destruction; inflammatory cells and osteoclasts in group D was less than those in groups B and C, with no obvious bone destruction. Significant difference was found in the limiting membrane thickness and the number of osteoclasts (group A lt; group D lt; group B lt; group C, P lt; 0.05). Western blot showed that the TNF-α expression levels were 0.235 ± 0.022, 0.561 ± 0.031, 0.731 ± 0.037, and 0.329 ± 0.025 in groups A, B, C, and D respectively, showing significant difference among 4 groups (P lt; 0.05). Conclusion The recombinant adenovirus for silencing TNF-α is successfully constructed, which can effectively inhibit osteolysis by silencing TNF-α expression in the tissues around prosthesis in mice.
Objective To observe the effect of local injection of vascular endothel ial growth factor (VEGF) and VEGF antibody on the wear particle-induced osteolysis in the mouse air pouch model and to investigate the role of VEGF in the process of aseptic loosening of prosthesis. Methods The stem of metal hip prosthesis was obtained from the revision surgery.Metallic wear particles were made by vacuum ball mill ing. Wear particles suspension was prepared into the concentration of 10 mg/mL with PBS. Fifty female Kunming mice (aged 8-10 weeks, weighing about 25 g) were selected. Of 50 mice, 10 were used as the donors of bone graft, the other 40 were equally divided into control group (group A), particle group (group B), VEGF group (group C), and VEGF inhibited group (group D). Air pouches were made on the back of 40 mice by injecting sterile air subcutaneously. At 8th day, a graft of calvaria from the donor mice was implanted in air pouch. In groups B, C, and D, 0.5 mL wear particles suspension was injected into the air pouches, and in group A, 0.5 mL PBS was injected. Once a day at 6th and 7th days during the air pouch preparation and one time every two days after bone implantation, 0.2 mL recombinant human VEGF (rhVEGF) and VEGF antibody (Bevacizumab) were injected into the air pouches in groups C and D, respectively. In group A and group B, 0.2 mL sal ine was injected. Pouch tissues and bone were harvested at 2 weeks after bone implantation for HE staining, real-time fluorescent quantitative PCR and ELISA analyses. Results All mice survived to the end of experiment. The gross observation showed that there were mild redness, swell ing, and less neovascularization in air pouches in group A. There were obvious redness, swell ing, and more exudative and neovascularization in groups B, C, and D, most obvious in group C, the next in group B, then in group D. The histological and molecular biological analysis showed that inflammatory responses and osteolysis were obvious in group B and the pouch membrane thickness, the cell density, transforming growth factor α, interleukin 1β, and VEGF were significantly higher than those in group A (P lt; 0.05). The inflammatory responses and osteolysis were mostobvious in group C and the above-mentioned indexes were significantly higher than those in group B (P lt; 0.05). There were some inflammatory responses and osteolysis in group D, but the indexes were significantly lower than those in group B (P lt; 0.05) and were significantly higher than those in group A (P lt; 0.05). Conclusion VEGF can promote inflammatory responses and osteolysis in aseptic loosening of prosthesis. VEGF antibody can effectively inhibit wear particle-induced osteolysis.
Objective To investigate the serum levels of endostatin and vascular endothelial growth factor ( VEGF) at different therapy stages of mouse Lewis lung carcinoma, and elucidate the relation to the progress and prognosis of tumor. Methods Forty-four Lewis lung carcinoma-bearing C57BL/6 mice were randomly divided into 4 groups, ie. a non-therapy group, a chemotherapy group, a gene therapy group, and a combination therapy group ( chemotherapy plus gene therapy) . Eleven healthy mice were included as normal control group. Serum was collected on the 0th, 5th, 19th day after therapy for measurement of endostatin and VEGF by ELISA. The correlations were analysed between endostatin and VEGF levels in each group. Results ( 1) The serum endostatin levels had no significant difference in all groups on the 0th day,but increased significantly on the 5th day in the gene and combination therapy groups than those in other three groups ( all P lt;0. 01) . Then the endostatin level decreased on the 19th day in the gene and combination therapy groups, but still higher than those in the chemotherapy group and the normal group. ( 2 ) On the contrary, serum VEGF levels of the gene and combination therapy groups decreased significantly on the 5th day and increased little on the 19th day, which were both significant lower than those in chemotherapy group on the 5th and 19th day( all P lt; 0. 05) . There were significant diferences between the three therapy groups and the non-therapy group( all P lt;0. 05) . ( 3) Negative correlations between VEGF and endostatin levels were revealed in the gene and combination therapy groups ( r = - 0. 77 and - 0. 761 respectively) .Correlation was not found in the non-therapy and chemotherapy groups. Conclusion The serum levels of endostatin and VEGF might be used as monitoring indices of antiangiogenesis therapy.
ObjectiveTo investigate the effects of icariin (ICA) on serum bone turnover markers expressions and histological changes of cartilage and subchondral bone in mouse osteoarthritis (OA) model.MethodsEighty 8-week-old male C57BL/6J mouse were randomly divided into 8 groups (n=10). The OA model was established by anterior cruciate ligament transaction (ACLT). Group A: sham operation/early-stage normal saline administration; group B: sham operation/early-stage ICA administration; group C: ACLT/early-stage normal saline administration; group D: ACLT/early-stage ICA administration; group E: sham operation/late-stage normal saline administration; group F: sham operation/late-stage ICA administration; group G: ACLT/late-stage normal saline administration; group H: ACLT/late-stage ICA administration. Each animal received either ACLT or simply opening joint capsule, respectively. For groups B and D, ICA was given by gavage [10 mg/(kg·day)] on the first day after ACLT. For groups F and H, ICA was given with the same volume at 4 weeks after operation. The blood serum of the mouse was collected and prepared at 8 weeks after operation. Serum bone turnover markers and cytokines, including C-telopeptide of type I collagen (CTX), osteocalcin (OC), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), and IL-1β, were measured by ELISA. Tissue samples from the knee were stained by alcian blue/hematoxylin & orange G (AB/H&OG). Histological changes of cartilage and subchondral bone were observed and evaluated by Osteoarthritis Research Society International (OARSI) scoring system.ResultsComparison between each group with early-stage administration (groups A, B, C, and D): Compared with groups A and B, the levels of CTX and OC in group C were significantly reduced (P<0.05); the levels of IL-6, TNF-α, and IL-1β and OARSI score was significantly increased (P<0.05). Compared with group C, the levels of CTX and OC in group D were significantly increased (P<0.05); the level of IL-6 was significantly reduced (P<0.05); the levels of TNF-α and IL-1β were not changed (P>0.05), and OARSI score was significantly reduced (P<0.05). Histological observation showed that the tibial cartilage loss was significantly improved. Comparison between each group with late-stage administration (groups E, F, G, and H): Compared with groups E and F, the levels of CTX and OC in group G were significantly reduced (P<0.05); the levels of IL-6, TNF-α, and IL-1β and OARSI score were significantly increased (P<0.05). Compared with group G, the level of CTX in group H were increased (P<0.05); the levels of OC, IL-6, TNF-α, and IL-1β and OARSI score were not changed (P>0.05). Histological observation showed that the tibial cartilage loss had no changes after late-stage ICA administration.ConclusionICA plays protective effects on subchondral bone, hyaline, and calcified cartilage. Meanwhile, ICA can improve bone remodeling in subchondral bone of OA to some extent. The consistent changes of serum bone markers and pathological morphology suggest that early intervention of ICA on OA is more effective.
ObjectiveTo investigate the mechanism of G protein coupled receptor kinase interacting protein 1 (GIT1) affecting angiogenesis by comparing the differentiation of bone marrow mesenchymal stem cells (BMSCs) differentiated into endothelial cells between GIT1 wild type mice and GIT1 gene knockout mice.MethodsMale and female GIT1 heterozygous mice were paired breeding, and the genotypic identification of newborn mice were detected by PCR. The 2nd generation BMSCs isolated from GIT1 wild type mice or GIT1 gene knockout mice were divided into 4 groups, including wild type control group (group A), wild type experimental group (group A1), GIT1 knockout control group (group B), and GIT1 knockout experimental group (group B1). The cells of groups A1 and B1 were cultured with the endothelial induction medium and the cells of groups A and B with normal cluture medium. The expressions of vascular endothelial growth factor receptor 2 (VEGFR-2), VEGFR-3, and phospho-VEGFR-2 (pVEGFR-2), and pVEGFR-3 proteins were detected by Western blot. The endothelial cell markers [von Willebrand factor (vWF), platelet-endothelial cell adhesion molecule 1 (PECAM-1), and vascular endothelial cadherin (VE-Cadherin)] were detected by flow cytometry. The 2nd generation BMSCs of GIT1 wild type mice were divided into 4 groups according to the different culture media: group Ⅰ, primary cell culture medium; group Ⅱ, cell culture medium containing SAR131675 (VEGFR-3 blocker); group Ⅲ, endothelial induction medium; group Ⅳ, endothelial induction medium containing SAR131675. The endothelial cell markers (vWF, PECAM-1, and VE-Cadherin) in 4 groups were also detected by flow cytometry.ResultsWestern blot results showed that there was no obviously difference in protein expressions of VEGFR-2 and pVEGFR-2 between groups; and the expressions of VEGFR-3 and pVEGFR-3 proteins in group A1 were obviously higher than those in groups A, B, and B1. The flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group A1 than in groups A, B, and B1 (P<0.05), and in group B1 than in groups A and B (P<0.05); but no significant difference was found between groups A and B (P>0.05). In the VEGFR-3 blocked experiment, the flow cytometry results showed that the expressions of vWF, PECAM-1, and VE-Cadherin were significantly higher in group Ⅲ than in groupsⅠ, Ⅱ, and Ⅳ, and in group Ⅳ than in groups Ⅰ and Ⅱ (P<0.05); but no significant difference was found between groups Ⅰ and Ⅱ (P>0.05).ConclusionGIT1 mediates BMSCs of mice differentiation into endothelial cells via VEGFR-3, thereby affecting the angiogenesis.