白头翁皂苷B4对慢性肾功能衰竭大鼠肾组织的保护作用及其机制

doi:10.13481/j.1671-587x.20200116

摘要/Abstract

摘要： 目的：探讨白头翁皂苷B4对慢性肾功能衰竭（CRF）大鼠氧化应激、炎症状态和肾组织中细胞凋亡相关蛋白表达的影响，阐明白头翁皂苷B4保护CRF大鼠肾组织的作用机制。方法：75只雄性SD大鼠分为对照组、模型组、阳性对照组、低剂量白头翁皂苷B4组和高剂量白头翁皂苷B4组。采用腺嘌呤连续灌胃21 d建立CRF大鼠模型。造模成功后，阳性对照组大鼠给予0.5 mg·kg^-1地塞米松，低和高剂量白头翁皂苷B4组大鼠分别灌胃给予1和2 mg·kg^-1白头翁皂苷B4，连续给药28 d，对照组和模型组大鼠灌胃给予等量生理盐水。采用生化分析仪检测大鼠血清中血肌酐（Scr）和尿素氮（BUN）水平，试剂盒法检测大鼠肾组织中超氧化物歧化酶（SOD）活性和丙二醛（MDA）水平，ELISA法检测大鼠肾组织中白细胞介素1β（IL-1β）、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）水平，Western blotting法检测肾组织中磷脂酰肌醇3激酶/蛋白激酶B（PI3K/Akt）通路的激活及凋亡相关蛋白B淋巴细胞瘤2（Bcl-2）、B淋巴细胞瘤2相关X蛋白（Bax）和含半胱氨酸的天冬氨酸蛋白水解酶3（Caspase-3）蛋白表达水平。结果：与对照组比较，模型组大鼠血清中Scr和BUN水平明显升高（P<0.01），正常肾小球和肾小体数量明显减少，肾组织中SOD活性降低（P<0.01），MDA、IL-1β、IL-6和TNF-α水平明显升高（P<0.01），Bax和Caspase-3蛋白表达水平明显升高（P<0.01），磷酸化Akt（p-Akt）和Bcl-2蛋白表达水平明显降低（P<0.01）。与模型组比较，阳性对照组、低和高剂量白头翁皂苷B4组大鼠血清中Scr和BUN水平降低（P<0.01），正常肾小球和肾小体数量明显增加，肾组织中SOD活性升高（P<0.01），MDA、IL-1β、IL-6和TNF-α水平明显降低（P<0.01），Bax和Caspase-3蛋白表达水平明显降低（P<0.01），p-Akt和Bcl-2蛋白表达水平明显升高（P<0.01）。结论：白头翁皂苷B4可通过降低血清中Scr和BUN水平，降低氧化应激反应，减少炎症因子的释放，激活肾组织PI3K/Akt通路，对肾组织损伤起到保护作用。

关键词: 白头翁皂苷B4, 慢性肾功能衰竭, 氧化应激, 尿肌酐, 炎症因子, 细胞凋亡

Abstract: Objective: To investigate the effects of anemoside B4 on the blood oxygen stress,inflanmation state and the expressions of apoptosis-related proteins in kidney tissue of the rats with chronic renal failure(CRF), and to illuminate the mechanism of anemoside B4 in protecting the kidney tissue of the CRF rats. Methods: A total of 75 SD male rats were divided into control group, model group, positive control group, low dose of anemoside B4 group and high dose of anemoside B4 group.The CRF rat models were established by continuous administration of adenine for 21 d. After successful modeling, the rats in low and high doses of anemoside B4 groups were given 1 and 2 mg·kg^-1 anemoside B4 by gavage and the rats in positive control group were given 0.5 mg·kg^-1 dexamethasone,lasted for 28 d; the rats in control group and model group were given the same amount of normal saline. Biochemical analyzer was used to detect serum urine creatinine (Scr) and urea nitrogen (BUN) levels. The kit method was used to detect the activity of superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) in kidney tissue of the rats. ELISA was used to detect the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factro-α (TNF-α) in kidney tissue of the rats. Western blotting method was used to detect the activation of PI3K/Akt pathway and the expression levels of apopotosis-related proteins Bcl-2, Bax and Caspase-3 in kindey tissue. Results: Compared with control group,the serum Scr and BUN levels of the rats in model group were increased(P<0.01),the number of normal glomerulus and renal corpuscles were decreased,the activity of SOD in kidney tissue was decreased(P<0.01),the levels of MDA,IL-1β,IL-6 and TNF-α were increased(P<0.01),the expression levels of Bax and Caspase-3 proteins were increased(P<0.01),and the expression levels of p-Akt and Bcl-2 proteins were decreased(P<0.01).Compared with model group, the serum Scr and BUN levels of the rats in positive control group, low and high doses of anemoside B4 groups were significantly decreased (P<0.01), the number of normal glomerulus and renal corpuscles were increased significantly, the activities of SOD in kidney tissue were increased significantly (P<0.01),the levels of MDA, IL-1β, IL-6, and TNF-α were significantly decreased (P<0.01), the protein expression levels of Bax and Caspase-3 were decreased significantly (P<0.01), and the expression levels of p-Akt and Bcl-2 proteins were increased significantly (P<0.01). Conclusion: Anemoside B4 can protect the renal tissue injury by reducing the levels of Scr and BUN in serum, reducing oxidative stress response, reducing the release of inflammatory factors, and activating the PI3K/Akt pathway of renal tissue.

Key words: anemoside B4, chronic renal failure, oxidative stress enzyme, serum urine cratinine, inflammatory cytokine, apoptosis

中图分类号:

R692.5

郭向辉, 郑慧, 吴巍. 白头翁皂苷B4对慢性肾功能衰竭大鼠肾组织的保护作用及其机制[J]. 吉林大学学报(医学版), 2020, 46(01): 90-95.

GUO Xianghui, ZHENG Hui, WU Wei. Protetive effects of anemoside B4 on kidney tissue of rats with chronic renal failure and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2020, 46(01): 90-95.

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