Astragalus granule ameliorates renal injury in diabetic rats and patients with diabetic kidney disease

1 Astragalus granule with losartan alleviated STZ-induced renal injury in DKD rats Significantly reduced kidney/body weight ratios and BUN were observed in AG+losartan group and Losartan group compared to Model group (P<0.01)(Figure 1). Compared to the losartan group, the ACR were also significantly reduced in AG+losartan group (P<0.05)(Figure 2). The results of histopathological tests of the renal tissues at the end of the experiment are shown in Figures 3. Pathological renal lesion significantly worsened in the model group. AG+losartan and losartan relieved glomerular hypertrophy and mesangial expansion (Figure 3). 2 In comparison with controls, the expression of TLR4, MyD88 and p-NF-κB significantly increased in the model group (P<0.05). After 12 weeks of treatment with Astragalus granules and losartan, the expression of TLR4, MyD88 and p-NF-κB protein decreased in the AG+losartan group and Losartan group (P<0.05). Compared to the losartan group, TLR4, MyD88 and p-NF-κB were also significantly reduced in AG+losartan group. In the all groups the level of NF-κB was not statistically different (Figure 4). 3 Astragalus granulesinhibited STZ-induced TNF-α and IL-1β generation in DKD rats Renal inflammation plays a key role in promoting the occurrence and development of DKD. To explore whether Astragalus granules could affect STZ-induced inflammatory cytokines production, ELISA was performed to detect the levels of TNF-α and IL-1β in blood of DKD rats. An apparent upregulation was observed in the levels of TNF-α and IL-1β in blood of DKD rats, while astragalus granules and losartan treatment strikingly inhibited the increase of TNF-α and IL-1β in DKD rats (Figure 5). 4 Astragalus granules alleviated renal injury in DKD patients The study was conducted as a randomized, controlled, prospective study. Patients were recruited over a time period of six months. Overall, 60 DKD patients with control group (n=30) and AG+losartan group (n=30) were included. The characteristics are in Table 1. There were no significantly different between the control group and AG+losartan group (Table 1). With 12 weeks treatment, the ACR in control group and AG+losartan group were significantly lower than 12 weeks ago. Meanwhile, compared to the control group, the ACR were significantly reduced in AG+losartan group (P<0.05). Alanine transaminase (ALT), Aspartic transaminase (AST), Blood urea nitrogen (BUN), Serum creatinine (SCR), Serum uric Acid (SUA), Glomerular filtration rate (GFR) and Kalium (K) were similar among the groups (Table 2). 5 Astragalus granulesinhibited TNF-α and IL-1β generation in DKD patients. ELISA was performed to detect the levels of TNF-α and IL-1β in blood of DKD patients. With 12 weeks treatment, astragalus granules and losartan treatment strikingly inhibited the increase of TNF-α and IL-1β in DKD rats (P<0.05)(Figure 6). But there was no significantly different in control group.1 黄芪颗粒联合氯沙坦可减轻链脲佐菌素（STZ）诱导的糖尿病肾病（DKD）大鼠肾损伤。与模型组相比，黄芪颗粒+氯沙坦组（AG+losartan组）与氯沙坦组的肾体重比及血尿素氮（BUN）水平均显著降低（P<0.01）（图1）。相较于氯沙坦组，黄芪颗粒+氯沙坦组的尿白蛋白/肌酐比值（ACR）同样显著降低（P<0.05）（图2）。实验结束时肾组织的病理组织学检测结果如图3所示。模型组大鼠肾脏病理损伤显著加重，黄芪颗粒+氯沙坦组与氯沙坦组均可缓解肾小球肥大与系膜扩张（图3）。 2 与对照组相比，模型组大鼠Toll样受体4（TLR4）、髓样分化因子88（MyD88）及磷酸化核因子-κB（p-NF-κB）的表达量显著升高（P<0.05）。经黄芪颗粒联合氯沙坦治疗12周后，黄芪颗粒+氯沙坦组与氯沙坦组大鼠的TLR4、MyD88及p-NF-κB蛋白表达量均显著下降（P<0.05）。相较于氯沙坦组，黄芪颗粒+氯沙坦组的TLR4、MyD88及p-NF-κB表达量进一步显著降低。所有组别中总核因子-κB（NF-κB）的水平均无统计学差异（图4）。 3 黄芪颗粒可抑制STZ诱导的DKD大鼠肿瘤坏死因子-α（TNF-α）与白细胞介素-1β（IL-1β）生成。肾脏炎症在DKD的发生与发展中发挥关键作用。为探究黄芪颗粒是否可影响STZ诱导的炎症因子产生，本研究采用酶联免疫吸附试验（ELISA）检测DKD大鼠血液中TNF-α与IL-1β的水平。结果显示，DKD大鼠血液中TNF-α与IL-1β的水平显著上调，而黄芪颗粒与氯沙坦治疗可显著抑制DKD大鼠体内TNF-α与IL-1β水平的升高（图5）。 4 黄芪颗粒可改善DKD患者的肾损伤。本研究为一项随机对照前瞻性研究，患者招募周期为6个月。最终纳入60例DKD患者，分为对照组（n=30）与黄芪颗粒+氯沙坦组（n=30）。两组患者的基线特征见表1，且组间无显著差异（表1）。经12周治疗后，对照组与黄芪颗粒+氯沙坦组的ACR均较治疗前显著降低；且相较于对照组，黄芪颗粒+氯沙坦组的ACR降低更为显著（P<0.05）。两组患者的丙氨酸氨基转移酶（ALT）、天冬氨酸氨基转移酶（AST）、血尿素氮（BUN）、血清肌酐（SCR）、血尿酸（SUA）、肾小球滤过率（GFR）及血钾（K）水平均无显著组间差异（表2）。 5 黄芪颗粒可抑制DKD患者体内TNF-α与IL-1β的生成。采用ELISA检测DKD患者血液中TNF-α与IL-1β的水平。经12周治疗后，黄芪颗粒与氯沙坦治疗可显著抑制DKD患者体内TNF-α与IL-1β水平的升高（P<0.05）（图6），但对照组无显著变化。