miR-21-3p/p53信号通路对缺血/再灌注损伤小鼠的肾脏保护作用及其机制

doi:10.3969/j.issn.1006-5725.2023.19.011

Abstract

Abstract:

Objective To observe the effect of miR-21-3p on mice with renal ischemia / reperfusion （I/R） injury and to explore whether the renal protective effect is related to the regulation of apoptosis levels through miR-21-3p/p53 signaling pathway. Methods In vivo experiments were randomly divided into five groups： Sham， I/R， I/R + agomir， I/R + agomir + PIF， and I/R + PIF. Renal I/R injury model in vivo was established， overexpressing of miR-21-3p （agomiR-21-3p） in mice were constructed and treated with the p53 inhibitor Pifithrin-α （PIF）. Dual-luciferase reporter assay was used to verify the interaction between miR-21-3p and p53. Serum levels of cystain C （Cys C）， blood urea nitrogen （BUN） and serum creatinine （Scr） were measured to reflect renal function of mice. Serum oxidative stress and inflammation indices were also determined， including tumor necrosis factor-alpha （TNF-α）， Interleukin-1β （IL-1β）， and high-sensitive c-reactive protein （hs-CRP）. In addition， apoptosis rate of renal cells was visualized by TUNEL staining and the expression of apoptosis b-cell lymphoma-2 （Bcl-2）， Bcl-2 assaciated X protein （Bax）， p53 and caspase-3 were measured too. Results Compared with Sham group， the expression of miR-21-3p in I/R mice was significantly decreased. Compared with I/R group， the expression of miR-21-3p in the I/R + agomir group， the I/R + PIF group and the I/R + agomir + PIF group was significantly increased （P < 0.05）， while the trend of p53 expression was opposite. Dual-luciferase reporter assay confirmed the interaction between miR-21-3p and p53. Compared with Sham group， the rate of cell apoptosis in all other renal tissues was increased； however， compared with the I/R group， the I/R + agomir， the I/R + PIF group and I/R + agomir + PIF groups were decreased， and the I/R + agomir + PIF group was the lowest （P < 0.05）. Conclusions miR-21-3p could inhibit the development of apoptosis by targeting p53 expression， thus alleviating renal injury in mice with renal I/R.

Key words: miR-21-3p, ischemia/reperfusion injury, p53, apoptosis

CLC Number:

R363.1+4

Xiaolin LEI,Yanxiu LIU,Chan. ZHANG. Protective effects and the mechanism of miR⁃21⁃3p/p53 signaling pathway on renal function in mice with renal ischemia/reperfusion injury[J]. The Journal of Practical Medicine, 2023, 39(19): 2475-2482.

Figures/Tables 7

Tab.1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

References 26

1	黄巍，张庆祥，罗丹，等. LncRNA MAGI2-AS3调控miR-374b-5p/HOXb7信号通路对脓毒症相关脑损伤的影响［J］. 实用医学杂志，2022，38（7）：815-820.
2	YANG J， SUN X， HUANG N， et al. Entacapone alleviates acute kidney injury by inhibiting ferroptosis［J］. FASEB J， 2022， 36（7）：e22399.
3	LIU Z， GUAN C， LI C， et al. Tilianin Reduces Apoptosis via the ERK/EGR1/BCL2L1 Pathway in Ischemia/Reperfusion-Induced Acute Kidney Injury Mice［J］. Front Pharmacol， 2022， 13：862584.
4	HOSOHATA K， JIN D， TAKAI S. In Vivo and In Vitro Evaluation of Urinary Biomarkers in Ischemia/Reperfusion-Induced Kidney Injury［J］. Int J Mol Sci， 2021， 22（21）：11448.
5	NI J， JIANG L， SHEN G， et al. Hydrogen sulfide reduces pyroptosis and alleviates ischemia-reperfusion-induced acute kidney injury by inhibiting NLRP3 inflammasome［J］. Life Sci， 2021， 284：119466.
6	GAUT J P， LIAPIS H. Acute kidney injury pathology and pathophysiology： a retrospective review［J］. Clin Kidney J， 2021， 14（2）：526-536.
7	ALLEGRETTI A S， SOLà E， GINèS P. Clinical Application of Kidney Biomarkers in Cirrhosis［J］. Am J Kidney Dis， 2020， 76（5）：710-719.
8	翁旭，武东辉，马文涛，等. lncRNA PCAT19靶向miR-769-5p促进口腔鳞癌HSC3细胞的增殖、迁移和侵袭［J］. 实用医学杂志，2023，39（4）：417-422.
9	姜平，郭哲，梁殿迅，等.长链非编码RNA X失活特异转录物调控miR-340-5p对卵巢癌细胞上皮间质转化的影响［J］. 实用医学杂志，2023，39（1）：41-47.
10	CAO J Y， WANG B， TANG T T， et al. Exosomal miR-125b-5p deriving from mesenchymal stem cells promotes tubular repair by suppression of p53 in ischemic acute kidney injury［J］. Theranostics， 2021， 11（11）：5248-5266.
11	LIU Z， YANG D， GAO J， et al. Discovery and validation of miR-452 as an effective biomarker for acute kidney injury in sepsis［J］. Theranostics， 2020， 10（26）：11963-11975.
12	HADDAD G， KöLLING M， WEGMANN U A， et al. Renal AAV2-Mediated Overexpression of Long Non-Coding RNA H19 Attenuates Ischemic Acute Kidney Injury Through Sponging of microRNA-30a-5p［J］. J Am Soc Nephrol， 2021， 32（2）：323-341.
13	PAN T， JIA P， CHEN N， et al. Delayed Remote Ischemic Preconditioning ConfersRenoprotection against Septic Acute Kidney Injury via Exosomal miR-21［J］. Theranostics， 2019， 9（2）：405-423.
14	LI X， LIAO J， SU X， et al. Human urine-derived stem cells protect against renal ischemia/reperfusion injury in a rat model via exosomal miR-146a-5p which targets IRAK1［J］. Theranostics， 2020， 10（21）：9561-9578.
15	ZHANG Y， LI C， GUAN C， et al. MiR-181d-5p Targets KLF6 to Improve Ischemia/Reperfusion-Induced AKI Through Effects on Renal Function， Apoptosis， and Inflammation［J］. Front Physiol， 2020， 11：510.
16	JIANG Y， XIE H， TU W， et al. Exosomes secreted by HUVECs attenuate hypoxia/reoxygenation-induced apoptosis in neural cells by suppressing miR-21-3p［J］. Am J Transl Res， 2018， 10（11）：3529-3541.
17	LIU R， ZHAO G， WANG Q， et al. Prognostic value of pulmonary ultrasound score combined with plasma miR-21-3p expression in patients with acute lung injury［J］. J Clin Lab Anal， 2022， 36（3）：e24275.
18	TANG C， MA Z， ZHU J， et al. P53 in kidney injury and repair： Mechanism and therapeutic potentials［J］. Pharmacol Ther， 2019， 195：5-12.
19	ZHANG C， LIANG R， GAN X， et al. MicroRNA-384-5p/Beclin-1 As Potential Indicators For Epigallocatechin Gallate Against Cardiomyocytes Ischemia Reperfusion Injury By Inhibiting Autophagy Via PI3K/Akt Pathway［J］. Drug Des Devel Ther， 2019， 13：3607-3623.
20	SUTTON T A， HATO T， MAI E， et al. p53 is renoprotective after ischemic kidney injury by reducing inflammation［J］. J Am Soc Nephrol， 2013， 24（1）：113-124.
21	ZHANG J， BI J， REN Y， et al. Involvement of GPX4 in irisin's protection against ischemia reperfusion-induced acute kidney injury［J］. J Cell Physiol， 2021， 236（2）：931-945.
22	XIAO Z， HUANG Q， YANG Y， et al. Emerging early diagnostic methods for acute kidney injury［J］. Theranostics， 2022， 12（6）：2963-2986.
23	TANG C， HAN H， LIU Z， et al. Activation of BNIP3-mediated mitophagy protects against renal ischemia-reperfusion injury［J］. Cell Death Dis， 2019， 10（9）：677.
24	ZHANG Z， CHEN H， ZHOU L， et al. Macrophage‑derived exosomal miRNA‑155 promotes tubular injury in ischemia‑induced acute kidney injury［J］. Int J Mol Med， 2022， 50（3）：116.
25	XU Y， JIANG W， ZHONG L， et al. circ-AKT3 aggravates renal ischaemia-reperfusion injury via regulating miR-144-5p /Wnt/β- catenin pathway and oxidative stress［J］. J Cell Mol Med， 2022， 26（6）：1766-1775.
26	LIAO Y， PENG X， LI X， et al. CircRNA_45478 promotes ischemic AKI by targeting the miR-190a-5p/PHLPP1 axis［J］. FASEB J， 2022， 36（12）：e22633.

名称	序列
p53	Forward: 5′-CCATCTACAAGCAGTCACA-3′
p53	Reverse: 5′-CCATCTACAAGCAGTCACA-3′
miR-21-3p	Forward: 5′ACACTCCAGCTGGGTAGCTTATCAGACTGA-3′
miR-21-3p	Reverse: 5′-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGCTACAACT-3′
U6	Forward: 5′-CTCGCTTCGGCAGCACA-3′
U6	Reverse: 5′-AACGCTTCACGAATTTGCGT-3′
GAPDH	Forward: 5′-TCAACGGCACAGTCAAGG-3′
GAPDH	Reverse: 5′-ACTCCACGACATACTCAGC-3′

[1]	Shan LUO,Ying FENG,Dandan FAN,Wenxin ZHENG,Xingrong GUO,Xuzhi. RUAN. ANGPTL8 knockout reduces lipopolysaccharide⁃induced hepatic lipid deposition [J]. The Journal of Practical Medicine, 2024, 40(9): 1197-1203.
[2]	Wei HE,Liping LIU,Jingwei ZHUO,Xiaodong ZHANG,Tong YANG,Jubin. FENG. CCR5 blockade reduces tumor growth by inducing apoptosis and impairing immunosuppression of tumor microenvironment [J]. The Journal of Practical Medicine, 2024, 40(9): 1204-1210.
[3]	Wenxin LI,Minjun LU,Li LIN,Yueqin LIU,Xiaolan. ZHU. circRAF1 regulates the proliferation and apoptosis of human ovarian granulosa cells [J]. The Journal of Practical Medicine, 2024, 40(7): 910-917.
[4]	Zhen YANG,Shaoru JIANG,Xiaoyan CHEN,Xiaolin CHEN,Weimin DENG,Xinyu. GUO. Effect of Jinghou Zengzhi Granules on ovarian GDF9 secretion and granulosa cells apoptosis in controlled ovarian hyperstimulation rats [J]. The Journal of Practical Medicine, 2024, 40(7): 918-923.
[5]	Runwei MA,Chunjie MU,Wenting GUI,Yao DENG,Minzhang ZHAO,Min LIU,Yi SONG. LncRNA SENCR targeted miR⁃206 regulates proliferation and apoptosis of human vascular smooth muscle cells of aortic dissection tissues [J]. The Journal of Practical Medicine, 2024, 40(3): 302-308.
[6]	Lihong DING,Shijia GENG,Yujie WANG. Effects of wedelobata on apoptosis and secretion of inflammatory cytokines in the alveolar epithelium infected by Streptococcus pneumonia [J]. The Journal of Practical Medicine, 2024, 40(3): 316-320.
[7]	Bing′er WU,Qing LI,Kerong YANG,Jian ZHANG,Yi YU,Lei LEI,Bo. HU. Impact of spermidine on proliferation and apoptosis in diffuse large B⁃cell lymphoma cell lines [J]. The Journal of Practical Medicine, 2024, 40(22): 3130-3137.
[8]	Lili TANG,Xinyu WANG,Jie ZHANG,Yue ZHAO,Xiaoyue LI. Research progress on the relationship between m6A methylation modification and acute kidney injury [J]. The Journal of Practical Medicine, 2024, 40(2): 278-282.
[9]	Chunmei ZHANG,Xinhui HUANG,Jinqiu HU,Xiaoyan BI,Fuli. YA. Sulforaphane protects human platelets from high glucose⁃induced cellular apoptosis through down-regulating PI3K/Akt signaling pathway [J]. The Journal of Practical Medicine, 2024, 40(18): 2530-2536.
[10]	Rao LÜ,Jiadi YU,Liuzhen LI,Chulan ZHAN,Liyue ZHAO,Yueliang LI,Jun DONG,Jiao. LI. Molecular mechanism of young Sca⁃1 bone marrow stem cell on old cardiac fibroblast cell apoptosis in aging mice [J]. The Journal of Practical Medicine, 2024, 40(17): 2369-2374.
[11]	LI Xiaoping, ZHOU Hongjian, GAO Fangfang, LI Wei.. Tspan1 antagonizes oxaliplatin⁃induced apoptosis in colorectal cancer cells by inducing cellular autophagy [J]. The Journal of Practical Medicine, 2023, 39(9): 1072-1078.
[12]	HE Wanshan , HONG Xiaoshan, CHEN Guanqiao, CHEN Bin, WEN Bin, LIN Yu, WEI Zhifu, LUO Xiping. Screening of gene differential expression of NSUN2 in ovarian cancer cells based on transcriptome sequenc⁃ ing technology [J]. The Journal of Practical Medicine, 2023, 39(9): 1079-1085.
[13]	ZHANG Zhuoer, GAO Xiaoya, YU Juan, WANG Yuli.. Experimental study of lncRNA KCNQ1OT1 targeting miR⁃129⁃5p to regulate inflammation and apoptosis in a Parkinson′s disease cell model [J]. The Journal of Practical Medicine, 2023, 39(6): 672-678.
[14]	LI Mingjuan, LI Xuexin, LIU Li.. Effect of short⁃duration mechanical ventilation on diaphragm function in septic rats [J]. The Journal of Practical Medicine, 2023, 39(5): 579-590.
[15]	LIU Xueqing, GONG Xiuqun, WANG Xia, WANG Qiwei, LU Jun.. Therapeutic mechanism of edaravone dexboeol on vascular dementia [J]. The Journal of Practical Medicine, 2023, 39(4): 505-509.

Protective effects and the mechanism of miR⁃21⁃3p/p53 signaling pathway on renal function in mice with renal ischemia/reperfusion injury

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 7

References 26

Related Articles 15

Recommended Articles

Metrics

Comments