电针疗法对AKI犬的肾功能、钙磷代谢、抗氧化能力以及NRF2信号通路的影响

doi:10.11843/j.issn.0366-6964.2023.02.036

摘要/Abstract

摘要： 急性肾损伤（acute kidney injury，AKI）目前已成为犬最主要的一种肾系疾病，尤其在老年犬中发病率逐年上升。近年来，针刺疗法在宠物临床已得到广泛的应用，并逐渐从治疗瘫痪等外科疾病向治疗宠物内科疾病的方向发展。本研究以腺嘌呤为造模药物建立急性肾损伤犬模型，采用电针疗法进行治疗。研究电针疗法对于犬的肾功能、钙磷代谢、抗氧化能力以及对于NRF2通路相关蛋白的影响，探究电针疗法对犬的急性肾损伤的治疗作用。选取24只3~4 kg健康比格犬，随机分为对照组、造模组、常规治疗组、电针干预组、电针治疗组和联合治疗组。第1~15天为造模期，第16~30天为治疗期。试验结束后检测尿比重、血清中尿素氮（BUN）、肌酐（CREA）、尿酸（UA）、钙（Ca²⁺）和磷（P³⁺）的变化；影像诊断X光检测肾变化情况；检测血清及肾组织的超氧化物歧化酶（SOD）和丙二醛（MDA）；病理组织学观察各组试验犬肾组织病变的严重程度；采用qRT-PCR和Western blot方法检测各组试验犬肾组织中与抗氧化相关基因和蛋白的表达情况；用免疫组织化学的方法检测肾组织中与抗氧化相关蛋白在组织中的分布情况。结果显示：电针治疗后尿比重有所升高；血清生化结果显示，相比于腺嘌呤模型组，经电针治疗后的犬的血清BUN、CRE、UA有明显的降低，Ca²⁺明显增多，P³⁺明显降低；X光结果显示，电针治疗后肾大小有所减少；血液及肾组织SOD活力明显升高，MDA含量明显降低；HE染色结果显示，经过电针治疗后的犬的肾小管上皮细胞水肿程度减轻，坏死和空泡变性等病变也得到明显的改善；qRT-PCR和Western blot检测结果显示，与空白组相比，造模组KEAP1基因和蛋白表达上调，NRF2、HO-1、NQO1基因和蛋白表达下调。与造模组相比，各治疗组肾中KEAP1基因和蛋白表达下调，NRF2、HO-1、NQO1基因和蛋白表达上调，其中，联合治疗组的KEAP1基因和蛋白表达上的变化比其余治疗组更明显；电针干预组的NRF2蛋白表达上的变化比其余各治疗组更明显。通过应用中兽医电针疗法可以缓解犬的急性肾损伤症状，减轻肾损伤。其中，电针与常规疗法相结合的联合治疗方法对于急性肾损伤的治疗效果较好，其治疗效果优于单独使用电针治疗与单纯进行对症治疗。

关键词: 犬, 急性肾损伤, 中兽医学, 电针疗法, NRF2通路

Abstract: At present, acute kidney injury (AKI) has become the most important renal disease in canine, especially in the elderly canine with an increasing incidence year by year. Acupuncture therapy has been widely used in pet clinic in recent years, and has gradually developed from treating paralysis and other surgical diseases to treating pet medical diseases. In this study, adenine was used as the modeling drug to establish the dog model of acute kidney injury, and electroacupuncture was used to treat it. To investigate the effects of electroacupuncture (EA) on kidney function, calcium and phosphorus metabolism, antioxidant capacity and NRF2 pathway related proteins in dogs, and to explore the therapeutic effect of EA on acute kidney injury in dogs. Twenty-four healthy beagles (3-4 kg) were randomly divided into control group, model group, conventional treatment group, EA intervention group, EA treatment group and combined treatment group. The 1st to 15th day for the modeling period, and the 16th to 30th day for the treatment period. After trial period, the changes of urine specific gravity, serum urea nitrogen (BUN), creatinine (CREA), uric acid (UA), calcium (Ca²⁺) and phosphorus (P³⁺) were detected. Imaging diagnosis X-ray detection of renal changes; SOD and MDA in serum and kidney were detected. The severity of renal tissue lesion was observed by histopathology. qRT-PCR and WB were used to detect the expression of antioxidation-related genes and proteins in renal tissues of experimental dogs. The distribution of antioxidant-related proteins in renal tissues was detected by immunohistochemistry. Here, we found that urine proportion increased after electroacupuncture intervention. Serum biochemical results showed that compared with adenine model group, BUN, CRE, UA, Ca²⁺ significantly increased, P³⁺ significantly decreased in serum after electroacupuncture treatment. X-ray results showed that kidney size decreased after electroacupuncture treatment. SOD activity in blood and kidney tissue were significantly increased, while MDA content decreased significantly. The results of HE staining showed that the edema degree of renal tubular epithelial cells was reduced, necrosis and vacuolar degeneration were also significantly improved after electroacupuncture treatment. qRT-PCR and WB results showed that KEAP1 gene and protein expression were up-regulated and NRF2, HO-1, NQO1 gene and protein expression were down-regulated in the modeling group compared with the blank group. Compared with the modeling group, the expressions of KEAP1 gene and protein were down-regulated in all treatment groups, while the expressions of NRF2, HO-1 and NQO1 gene and protein were up-regulated. The changes of KEAP1 gene and protein expression in the combined treatment group were more obvious than those in the other treatment groups. The changes of NRF2 protein expression in electroacupuncture intervention group were more obvious than those in other treatment groups.Electroacupuncture therapy can relieve the symptoms of acute kidney injury and kidney damage in dogs. Among them, electroacupuncture combined with conventional therapy has a better therapeutic effect on acute kidney injury, and its therapeutic effect is better than electroacupuncture alone or symptomatic treatment alone.

Key words: canine, acute kidney injury, Traditional Chinese Veterinary Medicine, electroacupuncture therapy, NRF2 pathway

中图分类号:

S853.6

张嘉宾, 徐钊, 周光余, 张梦迪, 符杨, 刘佳琪, 周东海. 电针疗法对AKI犬的肾功能、钙磷代谢、抗氧化能力以及NRF2信号通路的影响[J]. 畜牧兽医学报, 2023, 54(2): 803-815.

ZHANG Jiabin, XU Zhao, ZHOU Guangyu, ZHANG Mengdi, FU Yang, LIU Jiaqi, ZHOU Donghai. Effects of Electroacupuncture Therapy on Renal Function, Calcium and Phosphorus Metabolism, Antioxidant Capacity and NRF2 Signaling Pathway in AKI Dogs[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(2): 803-815.

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