NLRP3炎症小体在大鼠单侧输尿管梗阻引起肾间质纤维化中的作用及其机制

doi:10.13481/j.1671-587X.20240301

Abstract

Abstract:

Objective To discuss the effect of NOD-like receptor protein 3 （NLRP3） inflammasome on the renal interstitial fibrosis in the unilateral ureteral obstruction （UUO） model rats， and to clarify its potential mechanism. Methods Thirty healthy male Wistar rats were randomly divided into sham operation group （n=6） and UUO group （n=24）. The rats in sham operation group underwent the dissection of the ureter without ligation， while the rats in UUO group were sacrificed on the 3rd， 7th， and 14th days after operation， and based on the treatment duration，the rats were divided into UUO 3 d group （n=8）， UUO 7 d group （n=8）， and UUO 14 d group （n=8）. HE staining and Masson staining were used to observe the pathomorphology of kidney tissue of the rats in various groups； reagent kits were used to detect the levels of malondialdehyde （MDA）， activities of superoxide dismutase （SOD）， and levels of hydroxyproline （HYP） in kidney tissue of the rats in various groups；immunohistochemistry method was used to detect the expression levels of α-smooth muscle actin （α-SMA） and transforming growth factor-β1 （TGF-β1） proteins in kidney tissue of the rats in various groups； Western blotting method was used to detect the expression levels of NLRP3 protein in kidney tissue of the rats in various groups. Results The HE staining results showed significant tubular dilation， interstitial edema， and widening， with increased infiltration of inflammatory cells， and shedding of epithelial cells was seen in parts of the tubular lumina of the rats in UUO group. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； compared with UUO 3 d group and UUO 7 d group， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. The Masson staining results showed that in UUO group， there was evident infiltration of inflammatory cells in the renal interstitium and a noticeable increase in fibrotic tissue proliferation； with the increasing of duration of UUO， some tubular structures disappeared， and the interstitial widened further with gradually increasing collagen deposition， particularly at the corticomedullary junction. Compared with sham operation group， the interstitial fibrosis scores of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）； and compared with UUO 3 d and UUO 7 d groups， the interstitial fibrosis score of the rats in UUO 14 d group was significantly decreased （P<0.05）. Compared with sham operation group， the levels of MDA in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significantly increased （P<0.05）， and the SOD activities were significantly decreased （P<0.05）. Compared with sham operation group， the levels of HYP in obstructed kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）；compared with UUO 3 d group， the level of HYP in obstructed kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）. The immunohistochemistry results showed that compared with sham operation group， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were significant increased （P<0.05）； compared with UUO 3 d and UUO 7 d groups， the expression levels of α-SMA protein in kidney tissue of the rats in UUO 14 d group was significantly increased （P<0.05）； compared with sham operation group，the expression levels of TGF-β1 protein in renal tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 3 d， UUO 7 d， and UUO 14 d groups were also significantly increased （P<0.05）； compared with UUO 3 d group， the expression levels of TGF-β1 protein in the tubular epithelial cells and renal tubule interstitial tissue of the rats in UUO 14 d group were significantly decreased（P<0.05）. The Western blotting results showed that compared with sham operation group， the expression levels of NLRP3 protein in kidney tissue of the rats in UUO 7 d and UUO 14 d groups were significantly increased （P<0.05）. Conclusion The NLRP3 inflammasome plays a critical role in renal fibrosis of the UUO rats，and its mechanism may be related to the increasing of oxidative stress and the increasing of expression level of TGF-β1 protein.

Key words: Nucleotide combined with structure of oligomerization domain receptor protein 3, Oxidative stress, Transforming growth factor β1, Unilateral ureteral obstruction, Renal interstitial fibrosis

CLC Number:

R692

Yingxin RUAN,Junya JIA,Zhanfei WU,Wenya SHANG,Pengyu ZHANG. Effect of NLRP3 inflammatome in renal interstitial fibrosis induced by unilateral ureteral obstruction in rats and its mechanism[J].Journal of Jilin University(Medicine Edition), 2024, 50(3): 587-595.

Figures/Tables 6

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References 27

1	BLACK L M， LEVER J M， AGARWAL A. Renal inflammation and fibrosis： a double-edged sword［J］. J Histochem Cytochem， 2019， 67（9）： 663-681.
2	HUMPHREYS B D. Mechanisms of renal fibrosis［J］. Annu Rev Physiol， 2018， 80： 309-326.
3	ZHOU T， LUO M C， CAI W， et al. Runt-related transcription factor 1 （RUNX1） promotes TGF-β-induced renal tubular epithelial-to-mesenchymal transition （EMT） and renal fibrosis through the PI3K subunit p110δ［J］. EBioMedicine， 2018， 31： 217-225.
4	HUANG R S， FU P， MA L. Kidney fibrosis： from mechanisms to therapeutic medicines［J］. Signal Transduct Target Ther， 2023， 8（1）： 129.
5	VILAYSANE A， CHUN J， SEAMONE M E， et al. The NLRP3 inflammasome promotes renal inflammation and contributes to CKD［J］. J Am Soc Nephrol， 2010， 21（10）： 1732-1744.
6	KE B， SHEN W， FANG X D， et al. The NLPR3 inflammasome and obesity-related kidney disease［J］. J Cell Mol Med， 2018， 22（1）： 16-24.
7	阮颖新，张鹏宇，林珊，等. 内质网应激相关凋亡途径参与单侧输尿管梗阻大鼠肾间质纤维化［J］. 中华肾脏病杂志， 2011， 27（5）： 357-362.
8	HORTELANO S， GONZÁLEZ-COFRADE L， CUADRADO I， et al. Current status of terpenoids as inflammasome inhibitors［J］. Biochem Pharmacol， 2020， 172： 113739.
9	ZHANG W J， LI K Y， LAN Y， et al. NLRP3 Inflammasome： a key contributor to the inflammation formation［J］. Food Chem Toxicol， 2023， 174： 113683.
10	LIN C， JIANG Z X， CAO L， et al. Role of NLRP3 inflammasome in systemic sclerosis［J］. Arthritis Res Ther， 2022， 24（1）： 196.
11	POTERE N， DEL BUONO M G， CARICCHIO R， et al. Interleukin-1 and the NLRP3 inflammasome in COVID-19： Pathogenetic and therapeutic implications［J］. EBioMedicine， 2022， 85： 104299.
12	ALLOATTI G， PENNA C， COMITÀ S， et al. Aging， sex and NLRP3 inflammasome in cardiac ischaemic disease［J］. Vascul Pharmacol， 2022， 145： 107001.
13	BRAGA T T， FORESTO-NETO O， CAMARA N O S. The role of uric acid in inflammasome-mediated kidney injury［J］. Curr Opin Nephrol Hypertens， 2020， 29（4）： 423-431.
14	ARANDA-RIVERA A K， SRIVASTAVA A， CRUZ-GREGORIO A， et al. Involvement of inflammasome components in kidney disease［J］. Antioxidants， 2022， 11（2）： 246.
15	HUANG G Z， ZHANG Y D， ZHANG Y Y， et al. Chronic kidney disease and NLRP3 inflammasome： Pathogenesis， development and targeted therapeutic strategies［J］. Biochem Biophys Rep， 2023， 33： 101417.
16	ARANDA-RIVERA A K， CRUZ-GREGORIO A， APARICIO-TREJO O E， et al. Redox signaling pathways in unilateral ureteral obstruction （UUO）- induced renal fibrosis［J］. Free Radic Biol Med， 2021， 172： 65-81.
17	WU M， HAN W X， SONG S， et al. NLRP3 deficiency ameliorates renal inflammation and fibrosis in diabetic mice［J］. Mol Cell Endocrinol， 2018， 478： 115-125.
18	KIM Y G， KIM S M， KIM K P， et al. The role of inflammasome-dependent and inflammasome-independent NLRP3 in the kidney［J］. Cells， 2019， 8（11）： 1389.
19	MISHRA S R， MAHAPATRA K K， BEHERA B P， et al. Mitochondrial dysfunction as a driver of NLRP3 inflammasome activation and its modulation through mitophagy for potential therapeutics［J］. Int J Biochem Cell Biol， 2021， 136： 106013.
20	ZHUANG Y B， YASINTA M， HU C Y， et al. Mitochondrial dysfunction confers albumin-induced NLRP3 inflammasome activation and renal tubular injury［J］. Am J Physiol Renal Physiol， 2015， 308（8）： F857-F866.
21	WU Y S， HE F， LI Y Q， et al. Effects of Shizhifang on NLRP3 inflammasome activation and renal tubular injury in hyperuricemic rats［J］. Evid Based Complement Alternat Med， 2017， 2017： 7674240.
22	FENG H， GU J L， GOU F， et al. High glucose and lipopolysaccharide prime NLRP3 inflammasome via ROS/TXNIP pathway in mesangial cells［J］. J Diabetes Res， 2016， 2016： 6973175.
23	LIU H X， ZHAO L， YUE L， et al. Pterostilbene attenuates early brain injury following subarachnoid hemorrhage via inhibition of the NLRP3 inflammasome and Nox2-related oxidative stress［J］. Mol Neurobiol， 2017， 54（8）： 5928-5940.
24	梁文杰，毕建成，许庆友. NLRP3炎症小体在慢性肾脏病的表达及中药的干预机制［J］. 中药药理与临床， 2016， 32（3）： 208-211.
25	ANDERS H J， SUAREZ-ALVAREZ B， GRIGORESCU M， et al. The macrophage phenotype and inflammasome component NLRP3 contributes to nephrocalcinosis-related chronic kidney disease independent from IL-1-mediated tissue injury［J］. Kidney Int， 2018， 93（3）： 656-669.
26	WANG W J， WANG X Y， CHUN J， et al. Inflammasome-independent NLRP3 augments TGF-β signaling in kidney epithelium［J］. J Immunol， 2013， 190（3）： 1239-1249.
27	BRACEY N A， GERSHKOVICH B， CHUN J， et al. Mitochondrial NLRP3 protein induces reactive oxygen species to promote Smad protein signaling and fibrosis independent from the inflammasome［J］. J Biol Chem， 2014， 289（28）： 19571-19584.

Group	n	Score of renal interstitial fibrosis
Group	n	HE staining	Masson staining
Sham operation	6	0.41±0.14	0.44±0.13
UUO 3 d	8	1.42±0.40^*	1.57±0.37^*
UUO 7 d	8	2.01±0.45^*	2.01±0.44^*
UUO 14 d	8	2.68±0.53^*△#	2.64±0.57^*△#

Group	n	Level of MDA[m_B/（μmol·g^－1）]	Activity of SOD[λ_B/（U·mg^－1）]
Sham operation	6	1.42±0.60	106.00±25.97
UUO 3 d	8	4.14±1.61^*	74.88±21.82^*
UUO 7 d	8	3.89±1.44^*	55.38±18.60^*
UUO 14 d	8	3.58±1.38^*	49.13±15.83^*

Group	n	α-SMA protein	TGF-β1 protein
Sham operation	6	0.32±0.11	0.31±0.12
UUO 3 d	8	0.99±0.39^*	1.42±0.40^*
UUO 7 d	8	1.65±0.50^*	1.84±0.57^*
UUO 14 d	8	2.47±0.72^*△#	2.20±0.68^*△

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Effect of NLRP3 inflammatome in renal interstitial fibrosis induced by unilateral ureteral obstruction in rats and its mechanism

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