紫癜肾煎剂对过敏性紫癜性肾炎大鼠PI3K/AKT及HIF-1α/VEGFA信号通路的影响

Effects of Zidianshen Decoction(紫癜肾煎剂)on PI3K/AKT and HIF-1a/VEGFA Signaling Pathway in HSPN Rats

目的 观察紫癜肾煎剂对过敏性紫癜性肾炎(Henoch Schonlein purpura nephritis, HSPN)模型大鼠的治疗作用,并基于PI3K/AKT和HIF-1α/VEGFA信号通路探讨其作用机制。方法 随机选7只SD大鼠分为正常对照组,其余大鼠应用“BSA+LPS+CCL4”联合干姜建立HSPN大鼠模型,12周后随机抽取正常对照组和造模组的大鼠各1只,取肾组织固定包埋,采用免疫荧光检测造模组大鼠肾组织中IgA沉积并伴有蛋白尿,正常组无上述表现,提示造模成功。将造模成功的HSPN大鼠模型随机分模型组、紫癜肾煎剂低、高剂量(6.10、24.40 g/kg)组和西药(3.93 mg/kg)组,每组6只。给药结束后,采用溴甲酚紫法测定尿蛋白浓度,计算24 h尿蛋白定量;各组大鼠肾组织病理和免疫荧光检测;采用Western blotting法检测SD大鼠肾组织PI3K、AKT、HIF-1α、VEGFA蛋白表达情况。结果 紫癜肾煎剂可以显著降低24 h尿蛋白(P<0.05),减少肾小球系膜区免疫复合物沉积;与模型组比较,中药方组和西药组大鼠肾组织PI3K、AKT、HIF-1α、VEGFA蛋白表达明显降低,差异均有统计学意义(P<0.05)。结论 紫癜肾煎剂能减少HSPN大鼠24 h蛋白尿,改善肾功能及肾组织病理损伤,延缓HSPN病情进展,其机制可能与调控PI3K/AKT和HIF-1α/VEGFA信号通路有关。

Objective To observe the therapeutic effect of Zidianshen Decoction(紫癜肾煎剂)on Henoch-Schonlein purpura nephritis(HSPN)model rats,and to explore the mechanism based on phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)and hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor A(VEGFA)signaling pathway.Meth-ods 7 SD rats were randomly selected and divided into a normal control group.The remaining rats were used to establish the HSPN rat model using“bovine serum albumin(BSA)+lipopolysaccharide(LPS)+CCl4”combined with dried ginger.After 12 weeks,one rat from the normal control group and one rat from the model group were randomly selected,and the renal tissue was fixed and embedded.Immunofluorescence detection showed that immunoglobulin A(IgA)deposition in the renal tissue of the model group rats was accompanied by proteinuria.The normal group did not have the above manifestations,indicating that the model was successful.The successfully established HSPN rat model was randomly divided into model group,low and high dose(6.10,24.4 g/kg)groups of Zidianshen Decoction,and western medicine(3.93 mg/kg)group,with 6 rats in each group.After administration,the concentration of urinary protein was measured using bromocresol violet method,and the 24-hour urinary pro-tein quantification was calculated.Pathological and immunofluorescence detection of renal tissue in each group of rats and the ex-pressions of PI3K,AKT,HIF-1αand VEGFA in kidney tissue of SD rats were detected by Western blotting.Results Zidianshen Decoction can significantly reduce 24-hour urinary protein(P<0.05).Compared with those of the model group,the expressions of PI3K/AKT and HIF-1α/VEGFA protein in kidney tissue of Zidianshen Decoction groups and western medicine group were significantly decreased,and the differences were statistically significant(P<0.05).Conclusion Zidianshen Decoction can reduce 24-hour proteinuria in HSPN rats,improve the renal function and pathological damage and delay the progression of HSPN.The mechanism may be related to the regulation of PI3K/AKT and HIF-1α/VEGFA signaling pathway.