养阴活血方对狼疮样小鼠肾组织BMP-7、TGF-β_1表达的影响

Impacts of Yangyin Huoxue Formulas on the Expression of BMP-7 and TGF-β_1 in the Renal Tissues of cGVHD Mice

目的观察养阴活血方对慢性移植物抗宿主(cGVHD)小鼠的干预作用及对肾组织骨形成蛋白-7(BMP-7)和转化生长因子-β_1(TGF-β_1)表达的影响,探讨其作用机制。方法在无菌条件下,分离DBA/2小鼠淋巴结、脾脏和胸腺,剪碎,玻片研磨,PBS冲洗过筛,洗涤离心,制成淋巴细胞悬液,分四次对(C57BL/6J×DBA/2)F1代杂交鼠造模组进行尾静脉注射淋巴细胞悬液制备cGVHD小鼠模型。另取6只F1代小鼠为正常组。21只成模小鼠随机分为雷公藤多苷组、中药组、模型组,分别予雷公藤多苷、养阴活血方及等体积生理盐水灌胃。1个月后测小鼠血清尿素氮(BUN)、肌酐(Cr)、24 h尿蛋白定量,行肾脏HE染色,免疫组化染色法测小鼠肾组织BMP-7、TGF-β_1的含量。结果与模型组比较,中药组能明显改善cGVHD小鼠BUN、Cr、24 h尿蛋白,并上调肾组织BMP-7的含量(P<0.01),下调肾组织TGF-β_1的含量(P<0.05),中药组与雷公藤多苷组比较差异也有统计学意义(P<0.05)。结论养阴活血方治疗狼疮性肾炎的可能机制之一是通过下调肾组织TGF-β_1并上调BMP-7的表达来实现的。

Objective To observe the intervention effects of yangyin huoxue formulas on cGVHD mice and the impacts on BMP-7 and TGF-β1 in the renal tissues and explore its effect mechanism. Methods Under aseptic condition,DBA/2 lymph node,spleen and thymus of mouse were separated,cut up,ground,rinsed and screened with PBS,and finial the lymphocyte suspension was made after washing and centrifugation. In four times,in the modeling group of（ C57BL/6j × DBA/2） F1 generation of hybrid mice,the lymphocyte suspension was injected in tail nails to prepare cGVHD mouse models. Of them,6 F1 generation mice were collected in the normal group. 21 model mice were randomized into a tripterygium glycosides group,a Chinese medicine group and a model group,and treated with the lavage of tripterygium glycosides,yangyin huoxue decoction and physical saline of the same dose successively. 1 month later,the levels of serum BUN,Cr and 24 h urinary proteinwere detected. The renal HE staining was given and the IHC staining method was used to determine the expressions of BMP-7 and TGF-β1 in the renal tissues. Results Compared with the model group,the kidney function and 24 h urinary protein of cGVHD mice was improved apparently in the Chinese medicine group. BMP-7 level was up-regulated（ P 〈0. 01） and TGF-β1 was down-regulated（ P〈 0. 05）. The differences were also significant statistically between the Chinese medicine group and tripterygium glycosides group（ P〈 0. 05）. Conclusion The mechanism of yangyin huoxue formulas in the treatment of lupus nephritis is possibly relevant with the down-regulation of TGF-β1 and up-regulation of BMP-7 in the renal tissue.