间充质干细胞调节LXA4对糖尿病肾病大鼠的影响

Effects of Bone Marrow Mesenchymal Stem Cells Regulating LXA4 on Diabetic Nephropathy Rats

目的探讨骨髓间充质干细胞(mesenchymal stem cell,MSC)对糖尿病肾病(diabetic nephropathy,DN)大鼠肾损伤的影响及其可能的作用机制。方法8周龄雄性SD大鼠,按照随机数字表法分为DN组(n=30)和正常对照组(n=6)。DN组大鼠腹腔注射链脲佐菌素(streptozotocin,STZ)诱导DN模型,进一步将其分为DN组、DN+MSC组、DN+LXA4(脂氧素A4)组、DN+LXA4+WRW4(LXA4拮抗剂)组、DN+MSC+WRW4组,每组6只大鼠;正常对照组大鼠则给予等量枸橼酸钠溶液。大鼠肾小球系膜细胞(HBZY-1)加入高糖培养72h后,对细胞进行分组,即正常对照组、高糖组、高糖+MSC组、高糖+LXA4组、高糖+LXA4+WRW4组、高糖+MSC+WRW4组。使用酶联免疫吸附试验(enzyme linked immunoadsordent assay,ELISA)和Western blot法检测炎症和纤维化相关指标[转化生长因子-β1(transforming growth factor-β1,TGF-β1)、白细胞介素(interleukin,IL)-6、IL-8、干扰素-γ(interferon-γ,IFN-γ)]。结果利用大鼠肾小球系膜细胞和大鼠DN模型,本研究证实了MSC和LXA4均能够减低高糖组细胞中炎症和纤维化指标(TGF-β1、IL-6、IL-8、IFN-γ)的表达;经WRW4处理后,上述作用被抑制。同时本研究还证实了经MSC和LXA4处理后,高糖组细胞和DN组大鼠模型中p-Smad2、p-Smad3、CXCL1和CXCR2的表达减低;经WRW4抑制后,p-Smad2、p-Smad3、CXCL1、CXCR2表达均减少。结论MSC干预可抑制DN中肾脏的纤维化和炎性因子表达,MSC对DN的这种肾脏保护作用可能是通过提高DN肾组织中LXA4的表达而实现的。

Objective To investigate the effects of bone marrow mesenchymal stem cell(MSC)on kidney injury in diabetic Nephropathy(DN)rats and its possible mechanism.Methods The 8-week-old male SD rats were randomly divided into DN group(30 rats)and normal control group(6 rats)by random number table.The DN model was induced by intrabitoneal injection of streptozotocin(STZ),and the rats in the DN group successfully modeled were divided into DN group,DN+MSC group,DN+LXA4(lipoxin A4)group,DN+LXA4+WRW4(LXA4 antagonist)group,DN+MSC+WRW4group,There were 6 rats in each group;the rats of normal group were given the same amount of sodium citrate solution.The rat mesangial cells(HBZY-1)were cultured with high glucose for 72hours,and divided into four groups:normal control group,high glucose group,high glucose+MSC group,high glucose+LXA4group,high glucose+LXA4+WRW4group,high glucose+MSC+WRW4group.Markers of inflammation and fibrosis[transforming growth factor-β1(TGF-β1),interleukin(IL)-6,IL-8,interferon-γ(IFN-γEffects of Bone Marrow Mesenchymal Stem Cells Regulating LXA4 on Diabetic Nephropathy Rats.)]were detected by enzyme linked immunoadsordent assay(ELISA)and Western blot.Results Using rat mesangial cells and rat DN model,this study confirmed that both MSC and LXA4 could reduce the expression of inflammatory and fibrotic markers(TGF-β1,IL-6,IL-8,IFN-γ)in high glucose group,and the above effects were inhibited by WRW4 treatment.At the same time,this study also confirmed that the expression of p-Smad2,p-Smad3,CXCL1 and CXCR2 in high glucose group and rat DN mouse model decreased after treatment with MSC and LXA4,and the expression of p-Smad2,P-Smad3,CXCLI and CXCR2decreased after inhibition by WRW4.Conclusion MSC intervention can inhibit renal fibrosis and inflammatory factor expression in DN,MSC the renal protective effect of DN may be by raising the DN LXA4 in renal tissue of expression.