Zhu Qin, Li Xiao-Hong, Chen Hong-Yu, Jin Qin-Yang
Department of Nephrology, Key Laboratory of Zhejiang Province, Management of Kidney Disease, Hangzhou Hospital of Traditional Chinese Medicine, Hangzhou, Zhejiang 310007, P.R. China.
Department of Cardiology, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, Zhejiang 310014, P.R. China.
Exp Ther Med. 2021 Sep;22(3):962. doi: 10.3892/etm.2021.10394. Epub 2021 Jul 7.
The Chinese decoction compound centella formula (CCF) is clinically effective against diabetic kidney disease (DKD), but the exact mechanism remains unclear. The present study aimed to investigate the effects of CCF on OxInflammation and silent information regulator 1 (SIRT1) levels in rats with streptozotocin (STZ)-induced diabetes. Sprague-Dawley rats were divided into CCF, losartan, diabetic control (DC) and normal control (NC) groups (n=7). Except for the NC, all subgroups of rats were fed a high-fat diet for 112 days and received a single intraperitoneal injection of 35 mg/kg STZ on day 29. All rats were sacrificed on day 112. High-performance liquid chromatography was performed to analyse asiaticoside, astragaloside and triptolide levels in CCF (0.3400, 0.0640 and 0.0001 mg/ml, respectively). Fasting blood glucose, urine protein-to-creatinine ratio, serum creatinine and blood urea nitrogen were quantified. Periodic acid Schiff staining, H&E staining and transmission electron microscopy were used to examine kidney pathological changes. The mRNA and protein expression levels of SIRT1 in renal tissues were analysed by reverse transcription-quantitative PCR, western blotting and immunohistochemistry. Oxidative stress was evaluated by measuring the levels of superoxide dismutase (SOD), malondialdehyde (MDA) and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) in renal tissues. TNF-α and NF-κB p65 subunit in renal tissues were assessed for inflammation. Compared with the rats in the NC group, the rats in the DC group exhibited renal injury with proteinuria, decreased expression levels of SIRT1 and SOD (P<0.01) and increased levels of MDA, NOX4, TNF-α and NF-κB p65 (P<0.01). CCF treatment reduced proteinuria (P<0.01), alleviated renal damage, decreased MDA, NOX4, TNF-α and NF-κB p65 levels (P<0.01), increased SOD levels (P<0.05) and increased SIRT1 mRNA and protein expression levels (P<0.01). The present study indicates that CCF effectively protects the kidney from diabetes by inhibiting OxInflammation and upregulating SIRT1.
中药复方积雪草配方（CCF）对糖尿病肾病（DKD）具有临床疗效，但其确切机制尚不清楚。本研究旨在探讨CCF对链脲佐菌素（STZ）诱导的糖尿病大鼠氧化应激炎症及沉默信息调节因子1（SIRT1）水平的影响。将Sprague-Dawley大鼠分为CCF组、氯沙坦组、糖尿病对照组（DC）和正常对照组（NC）（n = 7）。除NC组外，所有大鼠亚组均给予高脂饮食112天，并在第29天腹腔注射一次35 mg/kg STZ。所有大鼠均于第112天处死。采用高效液相色谱法分析CCF中积雪草苷、黄芪甲苷和雷公藤甲素的含量（分别为0.3400、0.0640和0.0001 mg/ml）。对空腹血糖、尿蛋白/肌酐比值、血清肌酐和血尿素氮进行定量分析。采用高碘酸希夫染色、苏木精-伊红染色和透射电子显微镜检查肾脏病理变化。通过逆转录定量PCR、蛋白质印迹法和免疫组织化学分析肾组织中SIRT1的mRNA和蛋白表达水平。通过检测肾组织中超氧化物歧化酶（SOD）、丙二醛（MDA）和烟酰胺腺嘌呤二核苷酸磷酸氧化酶4（NOX4）的水平评估氧化应激。评估肾组织中肿瘤坏死因子-α（TNF-α）和核因子-κB p65亚基的炎症情况。与NC组大鼠相比，DC组大鼠出现蛋白尿性肾损伤，SIRT1和SOD表达水平降低（P < 0.01），MDA、NOX4、TNF-α和核因子-κB p65水平升高（P < 0.01）。CCF治疗可降低蛋白尿（P < 0.01），减轻肾损伤，降低MDA、NOX4、TNF-α和核因子-κB p65水平（P < 0.01），提高SOD水平（P < 0.05），并增加SIRT1 mRNA和蛋白表达水平（P < 0.01）。本研究表明，CCF通过抑制氧化应激炎症和上调SIRT1有效保护肾脏免受糖尿病侵害。
