运动神经元生存蛋白基因敲除在顺铂致小鼠急性肾损伤中的作用  

Effects of survival motor neuron gene knockdown on acute kidney injury induced by cisplatin in mice

作　　者：钱晓倩 朱冬冬 林芙君[1] 蒋更如[1] Qian Xiaoqian;Zhu Dongdong;Lin Fujun;Jiang Gengru(Renal Division,Xin Hua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200092,China)

机构地区：[1]上海交通大学医学院附属新华医院肾内科,上海200092

出　　处：《中华肾脏病杂志》2022年第11期975-982,共8页Chinese Journal of Nephrology

基　　金：国家自然科学基金(82070697);上海市卫生健康委员会科研项目(201940255)。

摘　　要：目的探讨运动神经元生存蛋白(survival motor neuron,SMN)基因敲除在顺铂诱导的急性肾损伤(acute kidney injury,AKI)小鼠中的作用。方法构建顺铂诱导的AKI小鼠(C57BL/6)模型,将雄性8~10周龄体重22~24 gSMN+/+野生型小鼠及SMN基因敲除杂合子(SMN^(+/-))小鼠随机分为4组:SMN+/+生理盐水组(野生型空白对照组,n=5)、SMN^(+/-)生理盐水组(SMN基因敲除杂合子空白对照组,n=5)、SMN+/+顺铂组(野生型顺铂组,n=5)和SMN^(+/-)顺铂组(SMN基因敲除杂合子顺铂组,n=5)。腹腔注射20 mg/kg顺铂溶液或0.9%生理盐水,72 h后处死小鼠,收集血清及肾组织。采用实时荧光定量PCR和Western印迹法检测SMN mRNA和蛋白表达水平,采用肌氨酸氧化酶法和脲酶法分别测定血清肌酐和尿素氮水平,PAS染色观察肾组织病理改变,TUNEL免疫荧光检测细胞凋亡水平,Western印迹和免疫组化法检测细胞凋亡标志蛋白多腺苷二磷酸核糖聚合酶(PARP)和内质网应激标志蛋白CHOP的表达。结果与野生型小鼠相比,SMN^(+/-)小鼠SMN mRNA和蛋白表达水平均较低,且顺铂腹腔注射后,SMN mRNA和蛋白表达水平进一步降低(均P<0.05)。野生型顺铂组小鼠血清肌酐、血清尿素氮、肾小管损伤评分、肾组织TUNEL阳性细胞数目、PARP蛋白表达及CHOP蛋白表达均高于野生型生理盐水组(均P<0.05),且SMN^(+/-)顺铂组小鼠上述指标表达均高于野生型顺铂组小鼠(均P<0.05)。结论SMN基因敲除可进一步加重顺铂诱导的AKI,促进肾小管上皮细胞凋亡。SMN可能是AKI治疗潜在的干预靶点。Objective To investigate the role of survival motor neuron(SMN)gene knockout in mice with cisplatin-induced acute kidney injury(AKI).Methods A mouse model(C57BL/6)of cisplatin-induced AKI was constructed.Twenty male wild type(WT)and SMN^(+/-)mice weighing 22-24 g were randomly divided into four groups:WT mice with saline injection group(WT vehicle,n=5),SMN^(+/-)mice with saline injection group(SMN^(+/-)vehicle,n=5),WT mice with cisplatin injection group(WT cisplatin,n=5)and SMN^(+/-)mice with cisplatin injection group(SMN^(+/-)cisplatin,n=5).Mice were injected intraperitoneally with 20 mg/kg cisplatin or 0.9%saline.72 hours later,the mice were sacrificed,and serum and kidney tissues were collected.The real time PCR and Western blotting were used to measure the expression levels of SMN mRNA and protein.The sarcosine oxidation and urease method were used to measure serum creatinine(Scr)and blood urea nitrogen(BUN)levels.Renal pathologic changes were observed by PAS staining.TUNEL immunofluorescence assay was used to detect the level of apoptosis.Western blotting and immunohistochemistry were used to detect the protein expression levels of apoptosis index poly(ADP-ribose)polymerase(PARP)and endoplasmic reticulum stress index CHOP.Results Compared with WT mice,SMN mRNA and protein expression levels were lower in SMN^(+/-)mice,and the expression level of SMN mRNA and protein was further decreased after intraperitoneal cisplatin injection(all P<0.05).Compared with WT mice with saline injection group,WT mice with cisplatin injection group had higher levels of Scr,BUN,tubular damage scores,TUNEL positive cell numbers,PARP and CHOP,while the expression levels of above indexes in the SMN^(+/-)mice with cisplatin injection group were higher than those in the WT mice with cisplatin injection group(all P<0.05).Conclusions SMN gene knockout can aggravate renal pathological damage and apoptosis of renal tubular epithelial cell in cisplatin-induced AKI mice.SMN may be a potential therapeutic target of AKI.

关 键 词：顺铂 急性肾损伤 细胞凋亡 运动神经元生存蛋白 

分 类 号：R692[医药卫生—泌尿科学]