黄连素调节鞘氨醇激酶-1-磷酸鞘氨醇信号通路抗糖尿病小鼠肾损伤的研究

Berberine ameliorates diabetic mouse renal injury through inhibition of SphK1-S1P signaling pathway

目的研究黄连素对四氧嘧啶诱导的糖尿病小鼠肾损伤的保护作用及其对糖尿病小鼠肾脏鞘氨醇激酶-1-磷酸鞘氨醇(SphK-S1P)信号通路的抑制效应。方法四氧嘧啶诱导的糖尿病小鼠采用黄连素(300 mg.kg-1.d-1)灌胃给药12周,正常组和糖尿病组小鼠给予同体积的溶媒。采用Re-al-time PCR技术检测肾组织中SphK1、TGF-β1、FN、ColⅣ的基因;Western blot法检测肾脏组织中SphK1、FN、ColⅣ的蛋白表达;LC-MS/MS检测肾脏组织中SphK1活性和S1P含量。结果黄连素明显抑制糖尿病小鼠血糖,肾重/体重比、血尿素氮、血肌酐和24 h尿蛋白异常增高;抑制肾脏肥大、纤维连接蛋白和Ⅳ型胶原积聚。此外,黄连素明显减少肾脏SphK1活性、mRNA和蛋白表达,降低S1P的生成。结论黄连素发挥抗糖尿病小鼠肾损伤的作用可能与抑制肾脏SphK-S1P信号通路的激活相关联。

Aim To evaluate the protective effects of berberine on alloxan-induced diabetic renal injury in mice and the inhibitory effects of berberine on the Sphingosine kinase Sphingosine 1-phosphate（SphK S1P） signaling pathway in mouse kidney.Methods Diabetic mice in berberine treatment group were treated orally with berberine（300 mg·kg-1·d-1） or vehicle for 12 weeks.Mice in control or diabetic group were administrated with the equal volume of vehicle.Western blot method was applied to detect the protein expression of SphK1,FN,Col IV and RT-PCR was used to detect the mRNA expression of SphK1,TGF-β1,FN,Col Ⅳ.Results Berberine significantly attenuated the abnormal increases in fasting blood glucose,kidney/body weight,blood urea nitrogen,serum creatinine and 24 h urine protein.In addition,berberine prevented renal hypertrophy,TGF-β1 synthesis,FN and Col Ⅳ accumulation in kidney of diabetic mouse.Moreover,berberine down-regulated the elevated stainining,activity and levels of mRNA and protein of SphK1,and S1P production as well.Conclusion The protective effects of berberine on the diabetic renal injury in mice are relevant to the inhibition of activation of SphK-S1P pathway in diabetic mouse kidney.