PIM1激酶在顺铂诱导的急性肾损伤中的作用

doi:10.19586/j.2095-2341.2022.0200

生物技术进展 ›› 2023, Vol. 13 ›› Issue (2): 298-304.DOI: 10.19586/j.2095-2341.2022.0200

• 研究论文 • 上一篇

PIM1激酶在顺铂诱导的急性肾损伤中的作用

李玉珍(), 朱杰夫, 吴雄飞()

武汉大学人民医院，武汉 430060

收稿日期:2022-11-24 接受日期:2022-12-16 出版日期:2023-03-25 发布日期:2023-04-07
通讯作者: 吴雄飞
作者简介:李玉珍 E-mail：llyz0321@163.com；
基金资助:
国家自然科学基金项目(82100803);湖北省自然科学基金项目(2021CFB101)

The Role of PIM1 Kinase in Cisplatin-induced Acute Kidney Injury

Yuzhen LI(), Jiefu ZHU, Xiongfei WU()

People’s Hospital of Wuhan University，Wuhan 430060，China

Received:2022-11-24 Accepted:2022-12-16 Online:2023-03-25 Published:2023-04-07
Contact: Xiongfei WU

摘要/Abstract

摘要：

PIM1是一种丝氨酸/苏氨酸激酶，参与多种信号通路调控，在许多疾病中发挥重要作用。为了探讨PIM1激酶在顺铂诱导的急性肾损伤中的表达情况及其作用，选取C57BL/6小鼠和肾小管细胞作为实验对象。采用腹腔注射顺铂构建顺铂诱导的急性肾损伤小鼠模型，通过检测肌酐尿素氮水平，HE染色检测肾组织病理变化，qRT-PCR和Western blot检测肾组织PIM1 mRNA和蛋白表达水平。腺病毒感染肾小管细胞构建PIM1过表达顺铂损伤模型，qRT-PCR和Western blot检测p62、Beclin-1和LC3B的表达水平，CCK-8以及流式细胞术评价肾小管细胞损伤情况。结果表明，在小鼠模型中，腹腔注射顺铂后血清肌酐和尿素氮水平显著升高，小鼠肾脏组织病理呈急性肾小管坏死状，肾组织中PIM1 mRNA与PIM1蛋白表达水平显著升高。在肾小管细胞模型中，PIM1过表达处理后，PIM1 mRNA及PIM1蛋白水平显著升高；顺铂处理后，肾小管细胞活性下降、细胞凋亡增加，过表达PIM1顺铂处理组较单纯顺铂处理组细胞活性增高，细胞凋亡率降低，细胞损伤减轻。顺铂处理后，肾小管细胞中p62蛋白表达减少，Beclin-1、LC3B蛋白表达增加，自噬激活；过表达PIM1顺铂处理组较单一顺铂处理组细胞p62蛋白表达增加，Beclin-1、LC3B蛋白表达减少，自噬减弱。研究结果说明，PIM1在Cis-AKI模型中被激活，过表达PIM1可以减轻顺铂导致的肾小管细胞损伤，这可能与其抑制细胞过度自噬有关。

关键词: PIM1激酶, 急性肾损伤, 顺铂, 自噬

Abstract:

PIM1 is a serine/threonine kinase that participates in the regulation of multiple signal pathways and plays an important role in many diseases. In order to explore the role of PIM1 kinase in cisplatin-induced acute kidney injury， C57BL/6 mice and renal tubular cells were selected as experimental objects. Cisplatin-induced acute kidney injury in mice was established by intraperitoneal injection of cisplatin. The levels of urea nitrogen and creatinine in serum were measured， renal pathology change was assayed by HE staining， qRT-PCR and Western blot were used to detect the expression levels of PIM1 mRNA and PIM1 protein. Renal tubular cells were infected by Adenovirus for overexpression of PIM1， the expression levels of p62、Beclin-1、LC3B were detected by Western blot， CCK-8 and flow cytometry were used to detect the renal tubular cell injury. Results showed that in vivo model， after injection of cisplatin， the levels of serum creatinine， urea nitrogen and tubular injury were significantly increased， the PIM1 mRNA and PIM1 protein expression in kidney tissues were dramatically increased. In vitro model， PIM1 mRNA and PIM1 protein expression levels significantly increased after Adenovirus infection. After cisplatin treated， the cell viability was decreased and the apoptosis was increased， while in the PIM1 overexpression group， the cell viability was increased and the apoptosis was decreased. After cisplatin treated， the expression level of p62 was decreased， the expression levels of Beclin-1， LC3B were increased， and autophagy was activated. Compared with the cisplatin treated group， the expression level of p62 was increased， the expression levels of Beclin-1， LC3B were decreased， and autophagy was weakened in the PIM1 overexpression group. In conclusion， PIM1 was activated in cisplatin-induced acute kidney injury， overexpression of PIM1 alleviated the cisplatin-induced renal tubular cells injury by inhibiting excessive autophagy.

Key words: PIM1 kinase, acute kidney injury, cisplatin, autophagy

中图分类号:

R459

李玉珍, 朱杰夫, 吴雄飞. PIM1激酶在顺铂诱导的急性肾损伤中的作用[J]. 生物技术进展, 2023, 13(2): 298-304.

Yuzhen LI, Jiefu ZHU, Xiongfei WU. The Role of PIM1 Kinase in Cisplatin-induced Acute Kidney Injury[J]. Current Biotechnology, 2023, 13(2): 298-304.

图/表 4

图1 小鼠Cis-AKI模型的建立A：小鼠AKI模型构建模式图；B：血清尿素氮水平；C：血清肌酐水平；D：肾组织HE染色（×400）及肾小管损伤评分；**表示与Saline组相比在P < 0.01水平上差异具有统计学意义。

Fig. 1 Establishment of mice Cis-AKI model

图2 PIM1在小鼠Cis-AKI肾组织中的表达情况A：qRT-PCR检测小鼠肾组织PIM1 mRNA表达水平；B：Western blot检测小鼠肾组织PIM1蛋白表达水平；**表示与Saline组相比在P < 0.01水平上差异具有统计学意义。

Fig.2 The expression of PIM1 in kidney tissue of mice Cis-AKI model

图3 过表达PIM1对顺铂导致的BUMPT细胞损伤的影响A：qRT-PCR检测BUMPT细胞中的PIM1 mRNA表达水平；B：Western blot 检测BUMPT细胞中PIM1蛋白表达水平；C：CCK8检测BUMPT细胞活性；D~E：流式细胞术检测BUPMT细胞凋亡；**表示与Saline组相比在P < 0.01水平上差异具有统计学意义（n=3）。

Fig. 3 The effect of PIM1 overexpression on the injury of BUMPT cells with cisplatin treated

图4 过表达PIM1对顺铂处理后BUMPT细胞中自噬相关蛋白表达水平的影响A~C：Western blot检测BUMPT细胞中自噬相关蛋白p62、Beclin-1、LC3B的表达。*，**分别表示两组处理间的差异在P<0.05和P<0.01水平上有统计学意义（n=3）。

Fig. 4 The effect of PIM1 overexpression on expression level of autophagy related proteins in BUMPT cells with cisplatin treated

参考文献 25

1	YANG Y, MENG L, WU S, et al.. LIGHT deficiency aggravates cisplatin-induced acute kidney injury by upregulating mitochondrial apoptosis[J/OL]. Int. Immunopharmacol., 2020, 89: 106999[2022-11-20]. .
2	HU J, GU W, MA N, et al.. Leonurine alleviates ferroptosis in cisplatin‐induced acute kidney injury by activating the Nrf2 signalling pathway[J]. Br. J. Pharmacol., 2022, 179(15): 3991-4009.
3	FANG C, LOU D, ZHOU L, et al.. Natural products: potential treatments for cisplatin-induced nephrotoxicity[J]. Acta. pharmacol. Sin., 2021, 42(12): 1951-1969.
4	孟利,杨雁,张克勤,等.Fgl2在顺铂诱导的小鼠急性肾损伤中的作用及机制研究[J].免疫学杂志,2021,37(2):93-99.
5	张术姣,谭小月,张勉之.丹参提取物对顺铂诱导小鼠急性肾损伤的影响及机制研究[J].中华中医药杂志,2021,36(12):7324-7328.
6	WANG B, HUANG C, LIU L, et al.. Pim1 kinase inhibitors exert anti-cancer activity against HER2-positive breast cancer cells through downregulation of HER2[J/OL]. Front. Pharmacol., 2021, 12: 614673[2022-11-22]. .
7	EBEID D E, FIROUZI F, ESQUER C Y, et al.. PIM1 promotes survival of cardiomyocytes by upregulating c-Kit protein expression[J/OL]. Cells, 2020, 9(9): 2001[2022-11-21]. .
8	ZHU H, WANG X, SUN Y, et al.. Pim1 overexpression prevents apoptosis in cardiomyocytes after exposure to hypoxia and oxidative stress via upregulating cell autophagy[J]. Cell Physiol. Biochem., 2018, 49(6): 2138-2150.
9	ANDREUCCI M, FAGA T, PISANI A, et al.. Quercetin protects against radiocontrast medium toxicity in human renal proximal tubular cells[J]. J. Cell Physiol., 2018, 233(5): 4116-4125.
10	SEIFERT C, BALZ E, HERZOG S, et al.. PIM1 inhibition affects glioblastoma stem cell behavior and kills glioblastoma stem-like cells[J/OL]. Int. J. Mol. Sci., 2021, 22(20): 11126[2022-11-22]. .
11	WU J, CHU E, KANG Y. PIM kinases in multiple myeloma[J/OL]. Cancers, 2021, 13(17): 4304[2022-11-22]. .
12	YUAN Y, WANG C, ZHUANG X, et al.. PIM1 promotes hepatic conversion by suppressing reprogramming-induced ferroptosis and cell cycle arrest[J]. Nat. Commun., 2022, 13(1): 5237-5356.
13	CHAUHAN S S, TOTH R K, JENSEN C C, et al.. PIM kinases alter mitochondrial dynamics and chemosensitivity in lung cancer[J]. Oncogene, 2020, 39(12): 2597-2611.
14	ZHAO B, LIU L, MAO J, et al.. PIM1 mediates epithelial-mesenchymal transition by targeting Smads and c-Myc in the nucleus and potentiates clear-cell renal-cell carcinoma oncogenesis[J/OL]. Cell Death. Dis., 2018, 9(3): 307[2022-11-22]. .
15	李侃,肖啸,周森,等.自噬途径在急性肾损伤中的作用及靶向治疗策略研究进展[J].中国综合临床,2022,38(5):466-470.
16	熊哲学,李凝旭,唐明娟.钠-葡萄糖协同转运蛋白2抑制剂对足细胞自噬影响及其肾脏保护机制的研究进展[J].中国糖尿病杂志,2022,30(7):554-557.
17	金丽霞,张晓东,潘超,等.中医药调控线粒体自噬防治慢性肾脏病的研究进展[J].中国中药杂志,2022,47(13):3432-3438.
18	KLIONSKY D J, PETRONI G, AMARAVADI R K, et al.. Autophagy in major human diseases[J/OL]. EMBO J., 2021, 40(19): e108863[2022-12-07]. .
19	KONG L, DENG J, ZHOU X, et al.. Sitagliptin activates the p62-Keap1-Nrf2 signalling pathway to alleviate oxidative stress and excessive autophagy in severe acute pancreatitis-related acute lung injury[J/OL]. Cell Death. Dis., 2021, 12(10): 928[2022-12-07]. .
20	JIANG K, XU Y, WANG D, et al.. Cardioprotective mechanism of SGLT2 inhibitor against myocardial infarction is through reduction of autosis[J]. Protein Cell, 2022, 13(5): 336-359.
21	王珍,杨洛,廖敏,等.mTOR信号通路在糖尿病肾病发病机制中的研究进展[J].生物技术进展,2021,11(3):316-321.
22	XU J, SHAN X, CHEN C, et al.. Tangshenning attenuates high glucose-induced podocyte injury via restoring autophagy activity through inhibiting mTORC1 activation[J/OL]. J. Diabetes. Res., 2022, 2022: 1610416[2022-12-08]. .
23	TAN X, ZHU H, TAO Q, et al.. FGF10 protects against renal ischemia/reperfusion injury by regulating autophagy and inflammatory signaling[J/OL]. Front. Genet., 2018, 9: 556[2022-11-22]. .
24	ZHOU J, FAN Y, ZHONG J, et al.. TAK1 mediates excessive autophagy via p38 and ERK in cisplatin‐induced acute kidney injury[J]. J. Cell Mol. Med., 2018, 22(5): 2908-2921.
25	ZHOU L, ZHANG L, ZHANG Y, et al.. PINK1 deficiency ameliorates cisplatin-induced acute kidney injury in rats[J/OL]. Front. Physiol., 2019, 10: 1225[2022-11-23]. .

[1]	赵云鹏, 张浩林, 熊倩倩, 李雨婷, 王娟. COPII囊泡衣被蛋白SEC24A在巨自噬通路中的功能研究[J]. 生物技术进展, 2022, 12(6): 906-914.
[2]	侯凯耀, 张二飞, 郑李娜, 陈红光, 谢克亮. 富氢液对脓毒症小鼠心肌细胞线粒体自噬的影响[J]. 生物技术进展, 2022, 12(4): 497-502.
[3]	王珍,杨洛,廖敏,郝亚荣. mTOR信号通路在糖尿病肾病发病机制中的研究进展[J]. 生物技术进展, 2021, 11(3): 316-321.
[4]	林晶晶,杨宇丰. 线粒体自噬的调控机制及其在相关疾病中的作用[J]. 生物技术进展, 2019, 9(5): 467-475.
[5]	徐长禄,赵艳红,马艺戈,王冰蕊,王鼎,郭青,佟静媛,高洁,李亚朴,刘金花,石莉红. 利用生物信息学方法解析自噬相关基因在人体红细胞终末分化各阶段的动态表达[J]. 生物技术进展, 2019, 9(3): 271-276.

PIM1激酶在顺铂诱导的急性肾损伤中的作用

The Role of PIM1 Kinase in Cisplatin-induced Acute Kidney Injury

RichHTML

PDF

可视化

摘要/Abstract

引用本文

使用本文

图/表 4

参考文献 25

相关文章 5

编辑推荐

Metrics

本文评价