当归红芪超滤膜提取物改善糖尿病肾病大鼠肾损伤的作用

Effects of ultra-filtration extract from Angelica Sinensis Radix and Hedysari Radix improves the kidney injury in diabetic kidney disease rats

目的探讨当归红芪超滤膜提取物改善糖尿病肾病(DKD)大鼠肾损伤可能的作用机制。方法用腹腔注射链脲佐菌素制备糖尿病模型,再用高糖高脂饲料喂养构建DKD模型。将造模成功的DKD大鼠分为模型组、阳性对照组和低、中、高剂量实验组,每组8只;另取8只Wistar大鼠作空白组。低、中、高剂量实验组分别灌胃给予1.5、3.0和6.0 g·kg^(-1)当归红芪超滤膜提取物溶液;阳性对照组灌胃给予1.75×10^(-3)g·kg^(-1)厄贝沙坦悬浊液;空白组和模型组均灌胃给予等体积纯净水。6组大鼠每天给药1次,连续给药12周。用考马斯亮蓝法测定24 h尿总蛋白(24 h-UTP),用蛋白质印迹法检测大鼠肾组织核因子红系2相关因子2(Nrf2)、血红素氧合酶1(HO-1)及长链脂酰辅酶A合成酶4(ACSL4)蛋白的表达水平。结果中、高剂量实验组和阳性对照组、模型组、空白组的24 h-UTP分别为(47.70±3.85)、(43.57±6.38)、(36.80±6.52)、(64.34±13.38)和(7.58±3.71)mg,Nrf2蛋白相对表达水平分别为0.86±0.08、0.75±0.06、0.64±0.08、1.09±0.06和0.60±0.07,HO-1蛋白相对表达水平分别为0.77±0.04、0.63±0.07、0.47±0.05、1.04±0.06和0.34±0.07,ACSL4蛋白相对表达水平分别为0.62±0.07、0.55±0.07、0.46±0.06、1.08±0.07和0.30±0.01。中、高剂量实验组和阳性对照组的上述指标与模型组比较,在统计学上差异均有统计学意义(P<0.05,P<0.01)。结论当归红芪超滤膜提取物可以改善DKD大鼠肾损伤,其机制可能与抑制Nrf2/HO-1通路过度激活导致的铁死亡有关。

Objective To explore the improve mechanism of ultrafiltration extract from Angelica Sinensis Radix and Hedy sari Radix on kidney injury in diabetic kidney disease(DKD)rats.Methods The diabetes model was prepared by intraperitoneal injection of streptozotocin,and then fed with high sugar and high fat diet.The rats with successful DKD were randomly divided into model group,positive control group and experimental-L,-M,-H groups with 8 cases per group.Additionally,selected 8 Wistar rats as the blank group.The experimental-L,-M,-H groups were given 1.5,3.0 and6.0 g·kg^(-1)ultra-filtration extract from Angelica Sinensis Radix and Hedysari Radix by gavage,respectively.The positive control group was given 1.75×10^(-3)g·kg^(-1)irbesartan suspension by gavage.The blank and model groups were given equal volume of pure water by gavage.Six groups were administrated once a day for 12 consecutive weeks.The 24 h-urinary total protein were detected by coomassie brilliant blue method.The protein expression levels of nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1)and andacyl-Co A synthetase long chain family member 4(ACSL4)in kidney tissue were detected by Western blot.Results The 24 h-UTP in the experimental-M,-H groups,positive control group,model group and blank group were(47.70±3.85),(43.57±6.38),(36.80±6.52),(64.34±13.38)and(7.58±3.71)mg;the relative expression levels of Nrf2 protein were 0.86±0.08,0.75±0.06,0.64±0.08,1.09±0.06 and 0.60±0.07;the relative expression levels of HO-1 protein were0.77±0.04,0.63±0.07,0.47±0.05,1.04±0.06 and 0.34±0.07;the relative expression levels of ACSLA protein were 0.62±0.07,0.55±0.07,0.46±0.06,1.08±0.07 and 0.30±0.01,respectively.Compared with the model group,the above indexes in the experimental-M,-H groups and positive control group were significantly different(P<0.05,P<0.01).Conclusion Ultra-filtration extract from Angelica Sinensis Radix and Hedysari Radix can improve the damage of kidney tissue in DKD rats,and the mechanism may be related to ferroptosis caused by excessive activation of Nrf2/HO-1 signaling pathway.