薯蓣皂苷元对顺铂诱导大鼠肾损伤的缓解作用

doi:10.16431/j.cnki.1671-7236.2022.09.037

摘要/Abstract

摘要： 【目的】探究薯蓣皂苷元（diosgenin，Dio）对顺铂（cisplatin，CP）诱导大鼠肾损伤的缓解作用。【方法】选取24只健康的雄性Wistar大鼠，随机分为对照组（C）、Dio组（Dio）、模型组（CP）和Dio干预组（Dio+CP）。C和CP组每天灌胃6 mL 0.5% CMC-Na，Dio和Dio+CP组每天灌胃Dio （60 mg/kg），连续灌胃10 d。在试验第7天，CP和Dio+CP组大鼠经尾静脉注射CP （6 mg/kg），然后每天继续灌胃Dio，CP给药72 h后处死各组大鼠，无菌分离肾脏组织，用HE染色观察肾脏病理变化；用试剂盒检测总抗氧化能力（T-AOC）、过氧化氢酶（CAT）活性及丙二醛（MDA）含量；用实时荧光定量PCR和Western blotting分别检测肾脏肿瘤坏死因子-α（TNF-α）、核因子κB p65（NFκB p65）、环氧合酶-2（COX-2）、白细胞介素-1β（IL-1β）和IL-10 mRNA和蛋白的表达；用免疫化学法检测肾脏受体相互作用蛋白激酶-1（RIPK1）和RIPK3蛋白的表达。【结果】 HE染色结果显示，对照组和Dio组肾脏结构正常；CP组大鼠肾小管上皮细胞发生脱落以及空泡变性，肾脏结构被破坏，Dio+CP组肾小管的损伤明显减轻。与对照组相比，CP和Dio+CP组肾脏中MDA含量极显著升高（P<0.01），T-AOC和CAT活性均极显著降低（P<0.01）；与CP组相比，Dio+CP组肾脏中MDA含量极显著降低（P<0.01），T-AOC和CAT活性均极显著增加（P<0.01）。实时荧光定量PCR和Western blotting结果表明，与对照组相比，CP组大鼠肾脏中TNF-α、COX-2、NFκB p65和IL-1β mRNA和蛋白的表达量均极显著升高（P<0.01），IL-10 mRNA和蛋白的表达量均极显著降低（P<0.01）；与CP组相比，Dio+CP组TNF-α、COX-2、NFκB p65和IL-1β mRNA和蛋白的表达量均显著或极显著降低（P<0.05；P<0.01），IL-10 mRNA和蛋白的表达量均极显著升高（P<0.01）。免疫组化结果表明，与对照组相比，CP组大鼠肾脏中RIPK1和RIPK3蛋白的表达量均极显著升高（P<0.01）；与CP组相比，Dio+CP组大鼠肾脏中RIPK1和RIPK3蛋白的表达量分别极显著（P<0.01）和显著（P<0.05）降低。【结论】 Dio通过抑制肾脏氧化应激、炎症反应和细胞程序性坏死缓解CP诱导的大鼠肾损伤。

关键词: 大鼠; 薯蓣皂苷元; 顺铂; 程序性坏死; 肾损伤

Abstract: 【Objective】 The aim of this study was to investigate the alleviating effect of diosgenin (Dio) on kidney injury induced by cisplatin (CP) in rats.【Method】 Twenty-four healthy male Wistar rats were randomly divided into control group (C), Dio group (Dio), model group (CP) and Dio intervention group (Dio+CP).C and CP groups were given a gavage of 6 mL 0.5% CMC-Na, Dio and Dio+CP groups were given a gavage of 60 mg/kg for 10 days.On day 7 of the experiment, CP and Dio+CP groups were injected with CP (6 mg/kg) through tail vein, and then Dio was gavaged continuously every day.After 72 h of CP administration, the rats in each group were sacrificed, and the renal tissues were separated aseptically, and the renal pathological changes were observed by HE staining.Total antioxidant capacity (T-AOC), catalase (CAT) activity and malondialdehyde (MDA) content were detected by kit.Renal tumor necrosis factor-α (TNF-α) and nuclear factor κB p65 (NFκB p65), cyclooxygenase-2(COX-2), interleukin-1 β (IL-1β) and IL-10 mRNA and protein expressions were detected by Real-time quantitative PCR and Western blotting respectively.The expressions of kidney receptor interacting protein kinase-1 (RIPK1) and RIPK3 protein were detected by immunochemistry.【Result】 HE staining results showed that the renal structure was normal in control and Dio groups.In CP group, the epithelial cells of the renal tubules were exfoliated and vacuolated, and the renal structure was destroyed.The damage of the renal tubules in Dio+CP group was significantly reduced.Compared with control group, the content of MDA was extremely significantly increased (P<0.01), the activities of T-AOC and CAT in CP and Dio+CP groups were extremely significantly decreased(P<0.01).Compared with CP group, the activities of T-AOC and CAT in kidney of Dio+CP group were extremely significantly increased (P<0.01), while the content of MDA was extremely significantly decreased (P<0.01).The results of Real-time quantitative PCR and Western blotting showed that compared with control group, the mRNA and protein expressions of TNF-α, COX-2, NFκB p65 and IL-1β, in the kidney of CP group were extremely significantly increased (P<0.01).The expressions of IL-10 mRNA and protein were extremely significantly decreased (P<0.01).Compared with CP group, the mRNA and protein expressions of TNF-α, COX-2, NFκB p65 and IL-1β in Dio+CP group were significantly or extremely significantly decreased (P<0.05; P<0.01), the mRNA and protein expression levels of IL-10 were extremely significantly increased (P<0.01).Immunohistochemical results showed that compared with control group, the expressions of RIPK1 and RIPK3 proteins in kidney of CP group were extremely significantly increased (P<0.01).Compared with CP group, the protein expressions of RIPK1 and RIPK3 protein in kidney of Dio+CP group were extremely significantly decreased (P<0.01) and significantly(P<0.05) decreased, respectively.【Conclusion】 Dio could alleviate CP-induced renal injury in rats by inhibiting renal oxidative stress, inflammatory response and programmed necrosis of cells.

Key words: rats; diosgenin; cisplatin; programmed necrosis; kidney injury

中图分类号:

S853

何炎峻, 金圣子, 王爽, 柳可欣, 刘云. 薯蓣皂苷元对顺铂诱导大鼠肾损伤的缓解作用[J]. 中国畜牧兽医, 2022, 49(9): 3633-3642.

HE Yanjun, JIN Shengzi, WANG Shuang, LIU Kexin, LIU Yun. Relaxation Effect of Diosgenin on Kidney Injury Induced by Cisplatin in Rats[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(9): 3633-3642.

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