Abstract: Objective To study the antiacute rejection effect of Pachymic acid (PA) in heart transplantation rats, in order to select a new antirejection medicine with low side effect from traditional Chinese medicine. Methods We established the model by transplanting Wistar rats (32,donor) heart allografts into the abdomen of SD rats (32,receptor). The homologous hearttransplanted rats were then randomly divided into 4 groups with 16 rats in each group. Olive oil solution with PA 1 mg/(kg·d), PA 10 mg/(kg·d), Cyclosporine (CsA) 5 mg/(kg·d) and olive oil solution 0.5 ml/(kg·d) were respectively given intragastrically to lowdosage PA group, highdosage PA group, CsA group and the control group till the end of observation. Survival time of heart allografts, heart beating and the histological changes of allografts were examined and serum level of interleukin2 (IL-2) and interferon-γ (IFN-γ) were determined by enzymelinked immunosorbent assay (ELISA). Results Survival time in the highdosage PA group, the lowdosage PA group and the CsA group were 24.90±0.99 d, 15.50±1.60 d and 26.80±0.88 d respectively, which is much better than the control group (6.10±1.10 d, q=22.363, P=0.000; q=44.793, P=0.000; q=49.272,P=0.000). IL-2 serum level in the highdosage PA group, the lowdosage PA group and the CsA group were all lower than that in the control group (q=14.483, P=0.000; q=3.705, P=0.000; =21.418,P=0.000), whileIL-2 serum level in the highdosage group was lower than that in the lowdosage group (q=10.778,P=0.000). Similarly, IFN-γ serum level in the first three groups were all lower than that in the control group (q=16.508,P=0000; q=4.281, P=0.000;q=19.621, P=0.000) and IFNγ serum level in the highdosage group was also lower than that in the lowdosage group (q=14.975, P=0.000). Pathological examination 7 days after the surgery showed that pathologic lesion was much more relieved in the two PA groups and the CsA group than the control group. Conclusion Acute rejection of heart transplantation can be effectively suppressed by PA.
Abstract: Objective To build a rat model of right ventricular failure （RVF） by subcutaneous injection of Monocrotaline. Methods Forty Wistar rats were equally divided into four groups, 10 rats each group. Exp4 group: four weeks after Monocrotaline injection, experimental results were observed; Exp6 group: six weeks after Monocrotaline injection, experimental results were observed; Con4 group: four weeks after normal saline injection, experimental results were observed; Con6 group: six weeks after normal saline injection, experimental results were observed. Four and six weeks after Monocrotaline or normal saline injection respectively, the hemodynamic indexes of each pair of groups were measured. Their hearts and livers were excised to measure physiological indexes and had pathological examinations. Results Mean pulmonary arterial pressure (MPAP), maximal rate of change of right ventricular pressure (RV dp/dtmax) and right ventricular ypertrophy index in Exp4 group were higher than those in Con4 group（Plt;0.05,0.01）. Compared with Con6 group, there were obvious symptoms of RVF in Exp6 group which included the increases of heart rate, increases of central venous pressure (CVP) and MPAP, the decreases of RV dp/dtmax, the decreases of weight, the increases of liver weight/body weight ratio and right ventricular hypertrophy index, significant pleural and peritoneal effusions（P＜0.05,0.01 ）. Pathological examination of Exp6 group showed disordering and bifurcated cardiac muscle fibers, large and thickly dying cell core, enlarged transverse diameter of the cardiac muscle fibers and stroma fibrosis. Vacuolar degeneration and dissolved carcoplasm could be seen. The vessel wall of the lung arteriole thickened, intercellular layer smooth muscle cell hyperplasied, elastic fibers increased, vessel wall arteriosclerosised, lumens stenosized. Conclusion This model is simple to build and successful rate is high. It is valuable for further research.
ObjectiveTo observe the effect of vascular endothelial growth factor/polylactide-polyethyleneglycol-polylactic acid copolymer/basic fibroblast growth factor (VEGF/PELA/bFGF) mixed microcapsules in promoting the angiogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. MethodsThe BMSCs were isolated by the method of whole bone marrow adherent, and sub-cultured. The passage 3 BMSCs were identified by Wright-Giemsa staining and flow cytometry, and used for subsequent experiments. VEGF/PELA/bFGF (group A), PELA/bFGF (group B), VEGF/PELA (group C), and PELA (group D) microcapsules were prepared. The biodegradable ability and cytotoxicity of PELA microcapsule were determined, and the slow-released ability of VEGF/PELA/bFGF mixed microcapsules was measured. The passage 3 BMSCs were co-cultured with the extracts of groups A, B, C, and D, separately. At 1, 3, 7, 14, and 20 days after being cultured, the morphological changes of induced BMSCs were recorded. At 21 days, the induced BMSCs were tested for DiI-labeled acetylated low density lipoprotein (Dil-ac-LDL) and FITC-labeled ulex europaeus agglutinin I (FITC-UEA-I) uptake ability. The tube-forming ability of the induced cells on Matrigel was also verified. The differences of the vascularize indexes in nodes, master junctions, master segments, and tot.master segments length in 4 groups were summarized and analyzed. ResultsThe isolated and cultured cells were identified as BMSCs. The degradation time of PELA was more than 20 days. There was no significant effect on cell viability under co-culture conditions. At 20 days, the cumulative release of VEGF in the mixed microcapsules exceeded 95%, and the quantity of bFGF exceeded 80%. The morphology of cells in groups A, B, and C were changed. The cells in groups A and B showed the typical change of cobble-stone morphology. The numbers of double fluorescent labeled cells observed by fluorescence microscope were the most in group A, and decreases from group B and group C, with the lowest in group D. The cells in groups A and B formed a grid-like structure on Matrigel. Quantitative analysis showed that the differences in the number of nodes, master junctions, master segments, and tot.master segments length between groups A, B and groups C, D were significant (P<0.05). The number of nodes and the tot.master segments length of group A were more than those of group B (P<0.05). There was no significant differences in the number of master junctions and master segments between group A and group B (P>0.05). ConclusionVEGF/PELA/bFGF mixed microcapsules have significantly ability to promote the angiogenic differentiation of rat BMSCs in vitro.