【目的】探讨吴茱萸碱调节鞘氨醇激酶1 （SphK1） /S1P信号通路对慢性肾衰竭 （CRF） 大鼠肾纤维化的影响。 【方法】通过 饲喂0.5%腺嘌呤饲料建立CRF大鼠模型。将造模后的大鼠随机分为模型组，吴茱萸碱低、高剂量组，尿毒清颗粒组及吴茱萸 碱高剂量+K6PC-5 （SphK1激活剂） 组，另设正常组。干预结束，检测24 h尿蛋白 （24 h-UTP） 及血清尿素氮 （BUN） 、血清肌酐 （SCr） 水平，酶联免疫吸附分析 （ELISA） 检测大鼠血清中单核细胞趋化因子1 （MCP-1） 、白细胞介素6 （IL-6） 水平，苏木素-伊 红 （HE） 染色法、Masson染色法观察肾组织病理学变化并计算胶原容积分数 （CVF） ，实时定量聚合酶链反应 （qRT-PCR） 法 检测肾组织中转化生长因子β1 （TGF-β1） 、Ⅳ型胶原 （COL-Ⅳ） mRNA表达，Western Blot法检测肾组织中SphK1、S1P蛋白 表达。 【结果】与正常组比较，模型组肾组织出现明显炎性浸润及胶原纤维沉积，肾小球数量减少，SCr、BUN、24 h-UTP， IL-6、MCP-1，CVF，TGF-β1、COL-Ⅳ mRNA，SphK1、S1P表达水平显著增加 （P＜0.05） ；与模型组比较，吴茱萸碱低、 高剂量组及尿毒清颗粒组肾组织病理改变得到改善，SCr、BUN、24 h-UTP，IL-6、MCP-1，CVF，TGF-β1、COL-Ⅳ mRNA，SphK1、S1P表达水平显著降低 （P＜0.05） ；各指标组间比较，吴茱萸碱低剂量组与吴茱萸碱高剂量组差异有统计学 意义 （P＜0.05） ，吴茱萸碱高剂量组与尿毒清颗粒差异无统计学意义 （P＞0.05） ；与吴茱萸碱高剂量组比较，吴茱萸碱高剂量 +K6PC-5组肾组织病理改变进一步加重，各指标的改善作用均被逆转 （P＜0.05） 。 【结论】吴茱萸碱可通过抑制SphK1/S1P信 号通路改善CRF大鼠肾纤维化。

Objective To investigate the effect of evodiamine on renal fibrosis in rats with chronic renal failure （CRF） by regulating sphingosine kinase 1（SphK1） /S1P signaling pathway. Methods CRF rat model was established by feeding 0.5% adenine diet. The rats after modeling were randomly divided into model group， evodiamine low- and high- dose groups，Niaoduqing Granules group and evodiamine high-dose+K6PC-5 （SphK1 activator） group. At the end of intervention，the levels of 24-hour urinary protein（24 h-UTP），serum urea nitrogen （BUN） and serum creatinine （SCr） were detected. The levels of monocyte chemoattractant protein 1 （MCP-1） and interleukin 6（IL-6） in serum of rats were detected by enzyme-linked immunosorbent assay（ELISA） . The pathological changes of renal tissue and collagen volume fraction （CVF） were observed by hematoxylin-eosin （HE） and Masson staining. The mRNA expressions of transforming growth factor β1（TGF- β1） and type Ⅳ collagen （COL-Ⅳ） in renal tissue were detected by real-time quantitative polymerase chain reaction （qRT-PCR） . The protein expressions of SphK1 and S1P in renal tissue were detected by Western Blot. Results Compared with the normal group，the model group showed obvious inflammatory infiltration and collagen fiber deposition，the number of glomeruli decreased in renal tissue，and the expression levels of SCr，BUN，24 h-UTP，IL-6，MCP- 1，CVF，mRNA expressions of TGF-β1 and COL-Ⅳ，protein expessions of SphK1 and S1P were significantly increased（P＜0.05） . Compared with the model group，the histopathological changes of the low-dose and high- dose evodiamine groups and the Niaoduqing Granules group was improved，and the expression levels of SCr， BUN，24 h-UTP，IL-6，MCP-1，CVF，mRNA expressions of TGF-β1 and COL-Ⅳ，protein expessions of SphK1 and S1P were significantly decreased. For intergroup comparison of each index，there were differences between the low-dose evodiamine group and the high-dose evodiamine group （P＜0.05）， but there was no significant difference between the high-dose evodiamine group and the Niaoduqing Granules group （P＞0.05） . Compared with the high-dose evodiamine group，the histopathological changes of the high-dose evodiamine + K6PC-5 group was further aggravated， and the improvement effect of all indexes were reversed （P＜0.05） . Conclusion Evodiamine improves renal fibrosis in CRF rats by inhibiting SphK1/S1P signaling pathway.

R285.5

国家自然科学基金资助项目 （编号：81860140）