长春新碱和环磷酰胺周期序贯联合对慢性移植物抗宿主病狼疮样小鼠肾脏病变的疗效研究

Effect and mechanisms of combined immunosuppressive agents sequential therapy in murine models of chronic graft-versus-host disease

目的通过周期序贯联合免疫抑制剂（长春新碱、环磷酰胺）治疗慢性移植物抗宿主病（cGVHD）狼疮样小鼠肾脏病变的实验研究，以阐明序贯联合用药对小鼠肾脏病变的治疗效果显著，且不良反应较小。方法选取cGVHD狼疮样小鼠模型30只，随机分为健康对照组、长春新碱间歇给药组、环磷酰胺隔日给药组、环磷酰胺间歇给药组、长春新碱＋环磷酰胺联合间隔给药组。经18周治疗后观察并检测。采用单因素方差分析和重复测量的方差分析进行统计学处理。结果①诱导造模后6周尿蛋白定量（24h）达（5．02±0．88）mg，抗双链DNA（dsDNA）抗体阳性，肾脏病理呈狼疮肾炎Ⅳ型表现，模型成功。②环磷酰胺隔日组[（0．48±0．32）mg]、长春新碱＋环磷酰胺联合组[（0．37±0．30）mg]对造模小鼠尿蛋白定量（24h）的改变最大，环磷酰胺间歇给药组、环磷酰胺隔日给药组与长春新碱＋环磷酰胺联合给药组对造模小鼠尿单核细胞趋化蛋白（MCP）-1值、尿转化生长因子（TGF）-β1值的降低幅度较大；而不同给药方案对血清MCP-1、TGF-β1的改变差异r无统计学意义（P〉0．05）。环磷酰胺隔日给药组、长春新碱＋环磷酰胺联合给药组造模小鼠肾脏MCP-1、TGF-β1表达在肾小球、肾小管中的阳性数最低，与对照组、长春新碱间歇给药组及环磷酰胺间歇给药组相比差异有统计学意义（P〈0.01）。③长春新碱和环磷酰胺周期联合对造模小鼠血丙氨酸转氨酶、血肌酐、血抗dsDNA抗体、尿蛋白定量（24h）、尿MCP-1、尿TGF-β1有交互作用（P〈0．05）。结论①序贯周期联合长春新碱和环磷酰胺联合治疗有交互作用，在降低尿蛋白定量（24h）、血清肌酐，减少肾脏MCP-1、TGF-β1的表达和排泌等方面有明显作用，与环磷酰胺隔日用药组相当，优于长春新碱间歇给药组、环磷酰胺间歇给药组。②长春新碱＋环磷酰胺联合间歇给药与单用药比较不良反应并没有增加。环磷酰胺隔日给药组的不良反应明显大于其他各组，表现为体质量不增、肝酶增高。

Objective To explore the effects and possible mechanisms of VCR combined with low dose cyclophosphamide （CTX） intermittently to treat severe systemic lupus erythematosus （SLE）. It is assumed that this might be a new combination therapy for SLE and expected to improve the overall prognosis and outcome of SLE. Methods Murine chronic graft-versus-host disease （cGVHD） model were developed for study. They were randomly divided into the control group, vincristine （VCR） pulse therapy group, CTX pulse therapy group, CTX every other day （EOD） group, VCR＋CTX combination group. One way ANOVA and repeated measure variance analysis were used for statistical analysis. Results （1） Six weeks after cGVHD models were set up, the average 24-hour urine protein quantification was （5.02±0.88） mg, anti-dsDNA antibody was positive, and 1V LN pathology could be observed histologically in the model murine. So cGVHD models were successfully developed. （1） Significantly difference in decreasing of 24-hour urine protein quantification was found in the CTX EOD group, VCR＋CTX combination group and other groups （P〈0.01）.Significant decrease in Cr, ALT, anti-dsDNA, was found in the CTX EOD group, VCR＋CTX combination group, CTX pulse therapy group and other groups （P〈0.05）. Decrease in urine MCP-1 and TGF-β1 could be detected, and statistical significant difference in these parameters could be found in the CTX EOD group, CTX pulse therapy group, VCR＋CTX combination group and other groups （P〈0.01）. MCP-1 and TGF-β1 expression in model kidney were reduced in the CTX EOD group, VCR＋CTX combination group and had statistical significant difference in the CTX EOD group, VCR＋CTX combination group, VCR pulse therapy group, and CTX pulse therapy group. （2） VCR and CTX combination treatment was effective in 24-hour urine protein quantification, blood Cr, ALT, anti-dsDNA and urine MCP-1, as well as urine TGF-β1 （P〈 0.05）. Conchmion （1） The combination of VCR and CTX is synergistic in decreasing 24-hour urine protein quantification, Cr, and the expression of MCP-1, TGF-β1） The adverse effect of VCR＋CTX combination group is similar to VCR pulse therapy group and CTX pulse therapy group.