经肾动脉移植不同剂量骨髓间充质干细胞对阿霉素肾病大鼠肾脏炎症的影响

Effect of transrenal artery transplantation of bone marrow mesenchymal stem cells with different doses on nephritis in adriamycin nephropathy rats

目的观察经肾动脉移植骨髓间充质干细胞(BMSCs)对阿霉素慢性肾脏病(CKD)大鼠的治疗效果,评估不同移植剂量对肾脏炎症的影响。方法雄性SD大鼠62只,2只4周龄大鼠用于体外分离培养BMSCs;48只6周龄大鼠造模(小剂量多次尾静脉注射阿霉素),成功后随机分组:模型组(M组)、低、中、高剂量组(A、B、C组),每组12只;A、B、C组经肾动脉移植BMSCs 0.5 mL,浓度分别为1×10^(6)个/mL、2×10^(6)个/mL、3×10^(6)个/mL;M组经肾动脉输注等量PBS,正常对照组(N组,12只),尾静脉输注等量0.9%NaCl。移植BMSCs后第7、14天检测血肌酐、尿素氮、24 h尿蛋白水平;制作肾脏病理切片行HE染色观察肾脏病理形态学改变,行免疫组化染色观察肾组织中炎症因子表达情况。结果与N组相比,M组、A组、B组、C组血肌酐、尿素氮、24 h尿蛋白均显著升高(P<0.05)。BMSCs移植7、14 d后,A组、B组、C组与M组比较血肌酐、尿素氮、24 h尿蛋白降低(P<0.05),且B、C组低于A组,但两组之间差异无统计学意义(P>0.05)。HE染色结果示:A组、B组、C组较M组萎缩肾小球数量减少,肾小管扩张情况得到改善,其中B、C组病理学形态较A组轻微;免疫组化结果示:A组、B组、C组大鼠肾间质CD3、CD20表达较M组表达减轻,其中B、C组减轻程度优于A组。结论经肾动脉移植BMSCs可抑制或延缓CKD大鼠肾脏炎症进展,且一定范围内,较大剂量效果优于小剂量。

Objective To observe the therapeutic of transrenal arterial transplantation of bone marrow mesenchymal stem cells(BMSCs)on Adriamycin-induced chronic kidney disease(CKD)rats,and to evaluate the different transplantation doses on renal inflammation.Methods Sixty-two male SD rats were selected as study objects,and 24-week-old male SD rats were used to separate BMSCs in vitro.Forty-eight 6-week-old rats were molded(intravenously injected with doxorubicin in small doses for several times),and randomly divided into model group(Group M),low-dose group(Group A),medium-dose group(Group B)and high-dose group(Group C),with 12 rats in each group.The rats in Group A,B and C received transrenal artery transplantation of BMSCs(0.5 mL),in which the concentrations were 1×10^(6)/mL,2×10^(6)/mL and 3×10^(6)/mL,respectively.Group M was infused with the same amount of PBS via renal artery,while the normal control group(Group N,12 rats)was infused with 0.9%NaCl through tail vein.The levels of serum creatinine,urea nitrogen and 24 h urinary protein were measured 7 and 14 days after BMSCs transplantation.HE staining was performed to observe the pathological changes of the kidney,and immunohistochemical staining was performed to analyze the expression of inflammatory cells.Results Compared with Group N,serum creatinine,urea nitrogen and 24 h urinary protein in Group M,A,B and C were significantly increased(P<0.05).After 7 and 14 days of BMSCs transplantation,serum creatinine,urea nitrogen and 24h urinary protein in Group A,B and C were significantly lower than those in Group M(P<0.05);which were lower in Group B and C than those in Group A,but there was no statistical significance(P>0.05).The results of HE staining showed that the numbers of atrophic glomerulus in Group A,B and C were lower than that in Group M,and the dilatation of renal tubules was also improved.The pathological morphology of Group B and C was slighter than that of Group A.Immunohistochemical results showed that the expression of CD3 and CD20 in renal interstitium of rats in Group A,B and C was lower than that in Group M,and it was less prominent in Group B and C than Group A.Conclusion Transrenal arterial transplantation of BMSCs can inhibit or delay the progression of renal inflammation in rats with CKD in dose-dependent manner in a certain range.