10-HDA对急性氯化镉所致肾损伤的保护作用及自噬相关蛋白表达的影响

Protective effect of 10-HDA on acute cadmium chloride-induced kidney damage and autophagy protein expression

[背景]急性镉暴露会引起多种组织器官损伤,肾脏是主要靶器官。镉诱导的急性肾损伤涉及多个复杂的机制,其中氧化应激和自噬起着重要作用。[目的]探讨10-羟基-2-癸烯酸(10-HDA)对氯化镉染毒小鼠急性肾损伤的影响,为镉中毒的发病机制和防治提供实验依据。[方法]将35只雄性C57BL/6小鼠按体重随机分为7组,每组5只。对照组腹腔注射生理盐水,染镉组腹腔注射4 mg·kg^(-1)氯化镉溶液,干预组腹腔注射4 mg·kg^(-1)氯化镉溶液同时灌胃50、100、150、200 mg·kg^(-1)的10-HDA,10-HDA组仅予以150 mg·kg^(-1)的10-HDA灌胃处理;连续处理14 d。最后一次染毒结束24 h后,通过检测血尿素氮、肌酐、丙二醛(MDA)、超氧化物歧化酶(SOD)等生理学指标,病理学指标,自噬相关蛋白(Atg7、Atg5、Beclin-1、LC3)和线粒体自噬相关蛋白(PINK1、Parkin)表达情况,检测10-HDA对氯化镉暴露所致肾损伤的作用。[结果]与对照组比较,染镉组小鼠体重明显降低(P<0.01);不同浓度的10-HDA干预后,小鼠体重较染镉组明显增长(P<0.01)。染镉组小鼠血尿素氮和肌酐水平均高于对照组(P<0.01);100、150、200 mg·kg^(-1)10-HDA干预后小鼠血尿素氮和肌酐含量较染镉组均有不同程度减少(P<0.01)。染镉组小鼠肾皮质MDA水平高于对照组,SOD活性低于对照组(P<0.01);不同剂量10-HDA干预组小鼠肾皮质MDA含量较染镉组有不同程度降低,SOD活性有不同程度增加(P<0.01),肾脏结构发生病理改变。氯化镉组Atg7、LC3-II/I表达高于对照组(P<0.05),而Beclin-1的表达降低(P<0.05);与氯化镉组相比,不同剂量10-HDA干预组的Atg7表达水平不同程度降减少,LC3-II/I表达在50、150、200 mg·kg^(-1)10-HDA干预组有不同程度减少,而Beclin-1的表达在50、100、150 mg·kg^(-1)10-HDA干预组增加(P<0.05)。染镉组和50 mg·kg^(-1)10-HDA干预组的PINK1和Parkin表达量低于对照组(P<0.01);与氯化镉暴露组相比,在100、150、200 mg·kg^(-1)10-HDA干预后,PINK1表达量均有不同程度的增加,Parkin表达量在所有10-HDA干预组均有所增加(P<0.01)。[结论]10-HDA处理可以减轻氯化镉引起的急性肾损伤,减少自噬相关蛋白的表达,增加线粒体自噬相关蛋白的表达。

[Background]Acute cadmium(Cd)exposure can cause damage to multiple tissues,with the kidney being the primary target organ.The development of Cd-induced acute kidney injury involves complex mechanisms,in which autophagy and oxidative stress play crucial roles.[Objective]To investigate the effect of 10-hydroxy-2-decenoic acid(10-HDA)on kidney injury in mice exposed to cadmium,and provide experimental basis for studying the pathogenesis and prevention of Cd poisoning.[Methods]Thirty-five male C57BL/6 mice were divided into 7 groups(each of 5 mice):control group(normal saline,intraperitoneal injection),CdCl2 group(4 mg·kg^(−1),intraperitoneal injection),intervention groups(4 mg·kg^(−1)CdCl2,intraperitoneal Injection+50,100,150,or 200 mg·kg^(−1)10-HDA,oral gavage),and 10-HDA group(150 mg·kg^(−1),oral gavage).All treatments were given for 14 d.Twenty-four hours after the last infection,physiological indicators[blood urea nitrogen(BUN),creatinine(CRE),malondialdehyde(MDA),and superoxide dismutase(SOD)],histopathological indicators,autophagy-related proteins(Atg7,Atg5,Beclin-1,and LC3),and mitochondrial autophagy-related proteins(PINK1 and Parkin)were detected to examine the effect of 10-HDA on kidney injury caused by CdCl2.[Results]Compared with the control group,the body weight of mice in the CdCl2 group was significantly reduced(P<0.01);compared with the CdCl2 group,the body weight of mice after intervention with different concentrations of 10-HDA was significantly increased(P<0.01).CdCl2 significantly increased BUN and CRE in the serum samples compared with the control group(P<0.01),which was significantly reduced to varying degrees after 100,150,and 200 mg·kg^(−1)10-HDA intervention(P<0.01).MDA significantly increased and SOD significantly decreased in the renal cortex following CdCl2 administration compared with the control group(P<0.01),which was resolved following 10-HDA administration at different concentrations(P<0.01).In histopathological studies,10-HDA restored injured kidney tissues induced by CdCl2.The expression levels of autophagy proteins Atg7 and LC3-II/I were significantly increased(P<0.05),and the expression level of Beclin-1 was significantly decreased(P<0.05)in the CdCl2 group compared with the control group.The expression levels of Atg7 were reduced to varying degrees after treatment with designed concentrations of 10-HDA,the expression levels of LC3-II/I were also reduced in the 50,150,and 200 mg·kg^(−1)10-HDA intervention groups,and the expression levels of Beclin-1 were increased in the 50,100,and 150 mg·kg^(−1)10-HDA intervention groups(P<0.05).The expression levels of PINK1 and Parkin in the CdCl2 group and the 50 mg·kg^(−1)10-HDA intervention group were lower than those in the control group(P<0.01).Compared with the CdCl2 group,the expression levels of PINK1 increased to varying degrees after 100,150,and 200 mg·kg^(−1)10-HDA intervention,and the expression levels of Parkin increased in all 10-HDA intervention groups(P<0.01).[Conclusion]The intervention using 10-HDA can lessen acute kidney injury caused by CdCl2,reduce the expression of autophagy-related proteins,and increase the expression of mitochondrial autophagy-related proteins.