祛湿化斑颗粒通过调控IL/STAT信号通路对过敏性紫癜性肾炎大鼠的影响

Effect of Qushihuaban granules on rats with Henoch-Sch nlein purpura nephritis by regulating IL/STAT signal pathway

探讨祛湿化斑颗粒(QSHB Gr)通过调控IL/STAT信号通路对过敏性紫癜性肾炎大鼠的影响及可能机制。将幼龄清洁(SPF)级SD大鼠36只随机分为6组:对照组、模型组、QSHB Gr/0.2剂低剂量组、QSHB Gr/0.4剂中剂量组、QSHB Gr/0.8剂高剂量组及霉酚酸酯(MMF 0.3 g/kg)组。综合模拟IgA肾病与血热证动物模型,复合成为过敏性紫癜肾炎大鼠(HSPN)模型;病理学检测肾脏损伤;测定血清肾脏生化指标总蛋白(TP)、白蛋白(ALB)的质量浓度以及肌酐(Cre)和血尿素氮(BUN)的浓度;收集24 h尿观察尿量并检测尿蛋白排泄情况;免疫组织化检测肾脏免疫复合物IgA、IgG沉积;Elisa法检测IL-17、IL-6、IL-2及TGF-β1的含量;Western blot检测STAT3、STAT5、RORγt、FoxP3蛋白表达。结果显示:与HSPN大鼠相比,QSHB Gr治疗可以通过减轻肾脏组织病理损伤、升高血清TP、ALB浓度,降低Cre、BUN浓度及24 h尿蛋白水平(P<0.01)显著改善HSPN大鼠肾脏损伤,且各个指标随QSHB Gr剂量增加变化越明显。QSHB Gr治疗可以显著抑制肾脏免疫复合物IgA、IgG沉积;显著降低IL-17、IL-6及TGF-β1的含量(P<0.05或P<0.01),升高IL-2含量(P<0.01);显著升高STAT5与FoxP3的表达水平(P<0.05或P<0.01),降低STAT3与RORγt的表达水平(P<0.01),各个指标随QSHB Gr剂量增加而变化越明显。以祛湿化斑颗粒(QSHB Gr)治疗可以改善过敏性紫癜性肾炎,可能是HSPN临床治疗的潜在候选药物。其机制可能与抑制IgA和IgG沉积及IL-6/STAT3与IL-2/STAT5信号通路调控密切相关。

The purpose of this essay is to explore the effect of Qushihuaban granules on rats with Henoch-Sch nlein purpura nephritis through regulating IL/STAT signal pathway and its possible mechanism.36 young SPF SD rats were randomly divided into 6 groups(n=6):Control group;Model group;QSHB Gr/0.2 low dose group;QSHB Gr/0.4 medium dose group;QSHB Gr/0.8 high dose group;Mycophenolate mofetil(MMF 0.3 g/kg)group.Comprehensively simulate the animal model of IgA nephropathy and blood-heat syndrome,and compound it into a rat model of Henoch-Sch nlein purpura nephritis(HSPN);pathologically detect kidney damage;determine serum kidney biochemical indicators,total protein(TP),the mass concentration of albumin(ALB),creatinine(Cre)and the concentration of blood urea nitrogen(BUN);collect 24 h urine to observe urine output and detect urine protein excretion;use immunohistochemistry to detect renal immune complex IgA and IgG deposition;use Elisa method to detect IL-17,IL-6,IL-2 and TGF-β1 content;Western blot detection of STAT3,STAT5,RORγt,FoxP3 protein expression.The results show that compared with HSPN rats,QSHB Gr treatment can significantly improve kidney damage in HSPN rats by reducing pathological damage of kidney tissue,increasing serum TP and ALB concentrations,reducing Cre and BUN concentrations and 24-hour urine protein levels(P<0.01)and each index changes more obviously with the increase of QSHB Gr dose.QSHB Gr treatment can significantly inhibit the deposition of renal immune complex IgA and IgG;significantly reduce the content of IL-17,IL-6 and TGF-β1(P<0.05 or P<0.01),and increase the content of IL-2(P<0.01);significantly increase the expression levels of STAT5 and FoxP3(P<0.05 or P<0.01),and decrease the expression levels of STAT3 and RORγt(P<0.01).Each index changes significantly with the increase of QSHB Gr dose.Therefore,QSHB Gr treatment can improve allergic purpura nephritis and may be a potential drug candidate for clinical treatment of HSPN.The mechanism may be closely related to the inhibition of IgA and IgG deposition and the regulation of IL-6/STAT3 and IL-2/STAT5 signaling pathways.