基于NF-κB通路研究生地黄-玄参对糖尿病肾病大鼠肾脏微炎症状态的影响

Effect of Rehmanniae Radix combined with Scrophulariae Radix on renal microinflammation in diabetic nephropathy rats based on NF-κB pathway

目的观察生地黄配伍玄参对DN大鼠病理过程中肾脏微炎症状态的影响。方法将50只SD大鼠适应性饲养1周后,随机选取10只作为空白对照组,其余采取STZ腹腔注射联合高脂饲料诱导DN模型,4周后将造模成功的大鼠按随机数字表法分为模型组、生地黄玄参组(5.25 g/kg)和二甲双胍组(200 mg/kg),每组10只。给药8周后,采用血糖仪检测大鼠空腹血糖;苯索氯铵比浊法检测尿微量白蛋白水平;ELISA法检测血清中胱抑素、TNF-α、IL-6、超敏C反应蛋白(hs-CRP)水平;采用HE、Masson、过碘酸希夫反应(PAS)染色观察肾脏病理形态改变;采用Western blot法检测肾组织单核细胞趋化蛋白-1(MCP-1)、NF-κB、p-NF-κB蛋白表达。结果与模型组比较,生地黄玄参组大鼠体重降低(P<0.05),空腹血糖[(18.06±5.69)mmol/L比(29.42±0.63)mmol/L]、24 h尿蛋白定量[(11.02±1.77)mg/d比(31.61±0.65)mg/d]、血清胱抑素[(208.16±12.07)ng/ml比(278.05±19.33)ng/ml]、TNF-α[(9.13±1.46)pg/ml比(73.16±8.30)pg/ml]、IL-6[(4.27±1.07)pg/ml比(12.04±1.57)pg/ml]、hs-CRP[(219.36±22.02)ng/ml比(266.97±15.80)ng/ml]水平降低(P<0.05),肾组织p-NF-κB[(0.49±0.07)比(0.84±0.12)]、MCP-1[(0.44±0.02)比(0.64±0.11)]蛋白表达降低(P<0.05)。结论生地黄配伍玄参可通过抑制NF-κB的过度活化,降低MCP-1相关蛋白表达,减轻肾脏微炎症状态,发挥保护肾脏、治疗DN的作用。

Objective To explore the effect of Rehmanniae Radix combined with Scrophulariae Radix on renal microinflammation in diabetic nephropathy(DN)rats.Methods 50 Sprague Dawley(SD)rats were adaptively fed for 1 week,and then 10 rats were randomly selected as the blank control group,and the rest were treated with STZ intraperitoneal injection combined with high-fat diet to induce DN model.After 4 weeks,the successful modeled rats were randomly divided into model group,Rehmannia glutinosa Scrophularia group(5.25 g/kg)and metformin group(200 mg/kg),with 10 rats in each group.After 8 weeks of administration,fasting blood glucose was measured by blood glucose meter;microalbuminuria was measured by benzalkonium chloride turbidimetry;serum cystatin,TNF-α,IL-6 and hs-CRP levels were measured by ELISA kit;renal pathological changes were detected by HE staining,Masson staining and PAS staining;the expression of MCP-1,NF-κB(total)and p-NF-κB protein in renal tissue was detected by Western blot.Results Compared with the model group,the body weight of rats in DHXS group was significantly decreased(P<0.05).The content of fasting blood glucose[(18.06±5.69)mmol/L vs.(29.42±0.63)mmol/L],24-hour urine protein[(11.02±1.77)mg/d vs.(31.61±0.65)mg/d],serum cystatin[(208.16±12.07)ng/ml vs.(278.05±19.33)ng/ml],TNF-α[(9.13±1.46)pg/ml vs.(73.16±8.30)pg/ml],IL-6[(4.27±1.07)pg/ml],hs-CRP[(219.36±22.02)ng/ml vs.(266.97±15.80)ng/ml]in DHXS group were significantly decreased(P<0.05),and the expression level of p-NF-κB(0.49±0.07 vs.0.84±0.12)and MCP-1(0.44±0.02 vs.0.64±0.11)in renal tissue of rats in DHXS group were significantly reduced(P<0.05).Conclusion Rehmanniae Radix combined with Scrophulariae Radix can protect kidney by inhibiting the over activation of NF-κB,and reducing the expression of MCP-1 related protein to reduce renal micro inflammation.