VEGF基因修饰大鼠羊膜间充质干细胞对肾病综合征大鼠血液生化指标的影响

Effect of VEGF gene-modified rat amniotic mesenchymal stem cells on blood biochemical parameters in rats with nephrotic syndrome

目的:探究血管内皮生长因子(VEGF)基因修饰大鼠羊膜间充质干细胞对肾病综合征大鼠血液生化指标的影响。方法:选取60只雄性大鼠,随机分为4组,各15例,分别为对照组(健康)、模型组(肾病综合征)、治疗1组(羊膜间充质干细胞悬液)、治疗2组(VEGF基因修饰羊膜间充质干细胞悬液)。观察4组肾组织病理形态学及羊膜间充质干细胞存活、分布情况,并比较尿肌酐(SCr)、血尿素(BU)、总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白(LDL)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、24 h尿蛋白定量(24 h UTP)、阻力指数(RI)、舒张期末期流速(EDV)、收缩期峰值流速(PSV)及肾组织VEGF相关表达。结果:对照组大鼠肾组织具有完整清晰结构,模型组大鼠肾组肾小球表现为肿胀,肾小球内存在较多细胞,系膜区明显增宽,系膜细胞及基质中到重度增生;相较于模型组,各治疗组病理改变均较轻,各治疗组病理改变程度均较模型组轻,其中治疗1组病理改变较治疗2组病理改变明显;对照组及模型组大鼠肾组织均未见荧光细胞;治疗1组、治疗2组大鼠肾组织均可见红色荧光的阳性细胞,且仅可见于肾间质及小管内,在肾小球中未见其表达。与治疗1组比较,治疗2组阳性细胞较多;与对照组比较,模型组SCr、BU、24 h UTP、TC、TG、LDL、IL-1β、TNF-α水平均明显升高(P<0.05),治疗1组、治疗2组SCr、BU、24 h UTP、TC、TG、LDL、IL-1β、TNF-α水平均较模型组明显降低,且治疗2组最低(P<0.05);与对照组比较,模型组RI明显升高,PSV、EDV明显下降(P<0.05),治疗1组、治疗2组PSV、EDV均较模型组显著升高,RI水平较模型组显著降低,且治疗2组RI最低,PSV、EDV最高(P<0.05);与对照组比较,模型组肾组织中VEGF蛋白及mRNA表达均明显下降(P<0.05),治疗1组、治疗2组肾组织中VEGF蛋白及mRNA表达均较模型组显著升高,且治疗2组最高(P<0.05)。结论:经VEGF基因修饰的羊膜间充质干细胞移植治疗,可显著改善肾病综合征大鼠血脂水平、微炎症状态、肾功能及肾动脉血流,效果更为显著,可为肾病综合征细胞移植治疗提供有力依据。

Objective:To investigate the effects of vascular endothelial growth factor(VEGF)gene modification of rat amniotic mesenchymal stem cells on blood biochemical parameters in rats with nephrotic syndrome.Methods:Sixty male rats were selected,and randomly divided into four groups(15 cases in each group):the control group(healthy),model group(nephrotic syndrome),treatment group 1(amniotic mesenchymal stem cell suspension),treatment group 2(VEGF gene-modified amniotic mesenchymal stem cell suspension).The renal tissue pathology,and survival and distribution of amniotic mesenchymal stem cells in four groups were observed,and the urine creatinine(SCr),blood urea(BU),total cholesterol(TC),triacylglycerol(TG),low-density lipoprotein(LDL),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),24 h urine protein quantification(24 h UTP),resistance index(RI),peak systolic flow velocity(PSV),end diastolic flow velocity(EDV)and expression of VEGF in renal tissue were compared among four groups.Results:In the control group,the renal tissue showed complete and clear structure,while in the model group,the glomerulus showed swelling,more cells in the glomerulus,the mesangial region widened significantly,and the mesangial cells and stroma were moderate to severe hyperplasia.Compared with the model group,the pathological changes of all treatment groups were lighter,and the degree of pathological changes of all treatment groups were lighter than that of model group,and the pathological changes of treatment group 1 was more obvious than that of the treatment group 2.No fluorescent cells were found in kidney tissue of rats in control group and model group.Red fluorescent positive cells were observed in renal tissue of the treatment group 1 and treatment group 2,and only in renal interstitium and tubule,but not in glomerulus.The positive cells in the treatment group 2 were more than that in treatment group 1.Compared with the control group,the levels of SCr,BU,24 h UTP,TC,TG,LDL,IL-1βand TNF-αin the model group significantly increased(P<0.05).The levels of SCr,BU,24 h UTP,TC,TG,LDL,IL-1βand TNF-αin the treatment group 1 and treatment group 2 were significantly lower than those in model group,and which was the lowest in treatment group 2(P<0.05).Compared with the control group,the RI significantly increased,the levels of PSV and EDV significantly decreased in the model group(P<0.05).Compared with the model group,the PSV and EDV in treatment group 1 and treatment group 2 significantly increased,the RI level significantly decreased,and the RI was the lowest and the PSV and EDV were the highest in the treatment group 2(P<0.05).Compared with the control group,the VEGF protein and mRNA expressions in the kidney tissue of model group significantly decreased(P<0.05).The VEGF protein and mRNA expressions in kidney tissue of treatment group 1 and treatment group 2 significantly increased compared with the model group,and which was the highest in the treatment group 2(P<0.05).Conclusions:VEGF gene modified amniotic mesenchymal stem cell transplantation can significantly improve the blood lipid level,microinflammatory state,renal function and renal artery blood flow in rats with nephrotic syndrome,which can provide a strong basis for cell transplantation therapy of nephrotic syndrome.