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Journal of Peking University (Health Sciences) ›› 2025, Vol. 57 ›› Issue (5): 875-883. doi: 10.19723/j.issn.1671-167X.2025.05.011

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Effect of aquaporin 5 on TLR4/MyD88/NF-κB signaling pathway in Sjögren syndrome rats

Lixiu ZHU1, Renli CHEN1,*(), Sujuan ZHOU2, Ye LIN1, Yirong TANG1, Zhen YE1   

  1. 1. Department of Rheumatology and Immunology, Ningde Hospital Affiliated to Ningde Normal University/Ningde Hospital Affiliated to Fujian Medical University, Ningde 352100, Fujian, China
    2. Department of Pathology, Ningde Hospital Affiliated to Ningde Normal University/Ningde Hospital Affiliated to Fujian Medical University, Ningde 352100, Fujian, China
  • Received:2023-05-05 Online:2025-10-18 Published:2025-07-25
  • Contact: Renli CHEN
  • Supported by:
    the Natural Science Foundation of Ningde(2022J09); the Ningde Normal University Joint University-Level Scientific Research Projects in the First Half of 2023(2023ZX702)

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Abstract: Objective: To investigate the effect of aquaporin 5 (AQP5) on Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor κB (NF-κB) signaling pathway in Sjögren syndrome (SS) rats. Methods: The SS gene expression data sets GSE406611 and GSE84844 were extracted from the Gene Expression Omnibus (GEO), and the AQP5 mRNA expression was analyzed by R software. The rat SS model was constructed. The successfully modeled rats were divided into SS group, SS+NC group, and SS+pc group, 10 rats in each group; and 10 rats were set as Normal group. The rats in the SS+NC group were injected with 10 μg of rno-pcDNA3.1-AQP5-NC at the submandibular gland, subcutaneously every day for 28 days. The rats in the SS+pc group were injected with 10 μg of rno-pcDNA3.1-AQP5 at the submandibular gland, subcutaneously every day for 28 days. The enzyme-linked immunosorbent assay (ELISA) kit was used to detect the content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the serum. High-throughput sequencing was used to identify the target genes. Quantitative real-time PCR (qPCR) and Western blot were used to detect the mRNA and protein expressions of AQP5, TLR4, MyD88, and NF-κB in the rat submandibular gland tissue. Results: In the SS dataset GSE406611 and GSE84844, the mRNA expression of AQP5 in SS was significantly reduced. Compared with the Normal group, the content of TNF-α and IL-1β in the serum, the mRNA and protein expressions of TLR4, MyD88, and NF-κB in the SS group were significantly increased, the mRNA and protein expressions of AQP5 were significantly decreased. After overexpression of AQP5, the content of TNF-α and IL-1β in the serum, the mRNA and protein expressions of TLR4, MyD88, and NF-κB in the SS+pc group were significantly decreased, the mRNA and protein expressions of AQP5 were significantly increased. The differences were statistically significant (all P < 0.05). Conclusion: The expression of AQP5 is involved in the progression of SS. Increasing the expression of AQP5 can significantly inhibit inflammatory stress and reduce the pathological damage of submandibular gland tissue. This may be related to the inhibition of TLR4/MyD88/NF-κB conduction.

Key words: Aquaporin 5, Sjögren syndrome, Toll-like receptor 4, Myeloid differentiation factor 88, NF-kappa B, Signaling pathway

CLC Number: 

  • R593.2

Table 1

Primer sequence of each gene"

Gene Sequence
rno-AQP5 Forward GGACCTGACCTGCCGTCTAG
Reverse TAGCCCAGGATGCCCTTGAG
rno-TLR4 Forward CCATGAGGCACATTGTTACG
Reverse AAGTGCTTCACCACCTGCTT
rno-MyD88 Forward CTAGCGACAAGCCATACACG
Reverse GTAGCCGAATCGTAGCCAGA
rno-NF-κB Forward GTGGGTTCAGATGAGGAGGA
Reverse TCTGGTCCAAATAGGCTTGG
rno-GAPDH Forward TGTAATAATTGTAGCCAAGTAAATCTCC
Reverse AAGTAACCATTTTTCAAAACATTCAAG

Figure 1

Online analysis of the expression of AQP5 gene in SS patients A, the volcanic diagram of differentially expressed genes and expression of AQP5 gene in dataset GSE406611; B, the volcanic diagram of differentially expressed gene and expression of AQP5 gene in dataset GSE84844. * P < 0.05. SS, Sjögren syndrome; AQP5, aquaporin 5."

Figure 2

Comparison of saliva secretion of rats in each group * P < 0.05, compared with the Normal group; # P < 0.05, compared with the SS group. SS, Sjögren syndrome; NC, negative control; pc, rno-pcDNA3.1-AQP5."

Figure 3

Comparison of submandibular gland tissue damage in each group of rats (HE staining ×400) SS, Sjögren syndrome; NC, negative control; pc, rno-pcDNA3.1-AQP5."

Figure 4

The comparison of the levels of TNF-α and IL-1β in serum of rats in each group * P < 0.05, compared with the Normal group; # P < 0.05, compared with the SS group. SS, Sjögren syndrome; NC, negative control; pc, rno-pcDNA3.1-AQP5; TNF-α, tumor necrosis factor-α; IL-1β, interleukin-1β."

Figure 5

Scatter plot of differentially expressed genes SS, Sjögren syndrome; NC, negative control; pc, rno-pcDNA3.1-AQP5."

Table 2

Top 10 genes of up-regulated and down-regulated expression"

Normal vs. SS SS+NC vs. SS+pc
Up-regulated Score Down-regulated Score Up-regulated Score Down-regulated Score
rno-TLR4 0.993 4 rno-AQP5 0.998 2 rno-AQP5 0.999 3 rno-TLR4 0.998 7
rno-MyD88 0.990 3 rno-ALOX15B 0.976 1 rno-AGTR1 0.976 4 rno-MyD88 0.973 4
rno-NF-κB 0.985 1 rno-ADCY7 0.962 5 rno-AGTR2 0.951 7 rno-NF-κB 0.950 5
rno-ATIC 0.963 7 rno-CACNA1S 0.948 9 rno-CALCA 0.934 8 rno-SPP1 0.893 9
rno-BDKRB1 0.952 9 rno-CAMK2B 0.930 3 rno-CAT 0.930 5 rno-SIRT1 0.891 6
rno-CCL2 0.941 8 rno-NCAM1 0.922 9 rno-NOS1 0.915 3 rno-TLR2 0.887 2
rno-CCR5 0.930 7 rno-NADK 0.917 7 rno-NOS2 0.909 1 rno-HTRIA 0.885 7
rno-IRF4 0.922 4 rno-MED1 0.901 4 rno-NR3C1 0.893 3 rno-GRM5 0.876 1
rno-JUN 0.913 8 rno-MC4R 0.899 0 rno-MTOR 0.877 2 rno-GNAT3 0.863 9
rno-LCK 0.892 9 rno-NCOA1 0.887 3 rno-KIT 0.861 7 rno-ESR1 0.852 8

Figure 6

The mRNA expression of rno-AQP5, rno-TLR4, rno-MyD88, and rno-NF-κB in the submandibular gland tissue of each group of rats * P < 0.05, compared with the Normal group; # P < 0.05, compared with the SS group. SS, Sjögren syndrome; NC, negative control; pc, rno-pcDNA3.1-AQP5; rno, Rattus norvegicus; AQP5, aquaporin 5; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation factor 88; NF-κB, nuclear factor κB."

Figure 7

Interaction network among AQP5, TLR4, MyD88, and NF-κB AQP5, aquaporin 5; TLR4, Toll-like receptor 4; MyD88, myeloid differentiation factor 88; NF-κB, nuclear factor κB; IRAK2, interleukin-1 receptor associated kinases 2; CD289, Toll-like receptor 9; TLR3, Toll-like receptor 3; IL1R1, interleukin 1 receptor type Ⅰ; TAB2, TAK1 binding protein 2; IL1RN, interleukin-1 receptor antagonist; PELI1, pellino-1; UBE2N, ubiquitin-conjugating enzyme E2 N; UBC, ubiquitin-conjugating enzyme; LY96, lymphocyte antigen 96; TICAM1, Toll interleukin-1 receptor domain containing adaptor molecule 1; TRAF3, tumor necrosis factor-associated factor 3; TIRAP, Toll interleukin-1 receptor domain containing adaptor protein; CD40, cluster of differentiation 40; TRAF6, tumor necrosis factor-associated factor 6; IRAK4, interleukin-1 receptor associated kinases 4; UBE2V1, ubiquitin conjugating enzyme E2 V1; MAP3K7, mitogen-activated protein kinase kinase kinase 7."

Figure 8

Expression of AQP5, TLR4, MyD88, and NF-κB in the submandibular gland tissue of each group of rats * P < 0.05, compared with the Normal group; # P < 0.05, compared with the SS group. Abbreviations as in Figure 6."

1
Pego-Reigosa JM, Restrepo Vélez J, Baldini C, et al. Comorbidities (excluding lymphoma) in Sjögren's syndrome[J]. Rheumatology (Oxford), 2021, 60(5): 2075- 2084.

doi: 10.1093/rheumatology/key329
2
Imgenberg-Kreuz J, Rasmussen A, Sivils K, et al. Genetics and epigenetics in primary Sjögren's syndrome[J]. Rheumatology (Oxford), 2021, 60(5): 2085- 2098.

doi: 10.1093/rheumatology/key330
3
李永明, 薛鸾. AQP5在干燥综合征发病机制中的作用[J]. 中华中医药学刊, 2019, 37(10): 2369- 2372.
4
Chivasso C, Hagströmer CJ, Rose KL, et al. Ezrin is a novel protein partner of aquaporin-5 in human salivary glands and shows altered expression and cellular localization in Sjögren's syndrome[J]. Int J Mol Sci, 2021, 22(17): 9213.

doi: 10.3390/ijms22179213
5
Soyfoo MS, Nicaise C. Pathophysiologic role of interleukin-33/ST2 in Sjögren's syndrome[J]. Autoimmun Rev, 2021, 20(3): 102756.

doi: 10.1016/j.autrev.2021.102756
6
Verstappen GM, Pringle S, Bootsma H, et al. Epithelial-immune cell interplay in primary Sjögren syndrome salivary gland pathogenesis[J]. Nat Rev Rheumatol, 2021, 17(6): 333- 348.

doi: 10.1038/s41584-021-00605-2
7
梅寒颖, 刘炬, 汤曾耀. 基于TLR4/MyD88/NF-κB信号通路探讨白芍总苷抑制干燥综合征模型小鼠炎症的作用机制[J]. 中药新药与临床药理, 2021, 32(9): 1293- 1299.
8
宋维海, 李琴, 茅建春. 一贯煎对干燥综合征模型大鼠及颌下腺PI3K/Akt/eNOS通路的影响[J]. 中药材, 2020, 43(7): 1721- 1725.
9
王信, 王健, 郭文静, 等. 原发性干燥综合征患者肠道菌群特点分析[J]. 南方医科大学学报, 2020, 40(7): 949- 957.
10
黄诗怡, 周广文, 朱伟, 等. 补肾化痰方对OVX诱导的骨质疏松大鼠血清LPS及TLR4/MyD88/NF-κB信号通路的影响[J]. 中国实验方剂学杂志, 2021, 27(9): 70- 76.
11
Juarez M, Diaz N, Johnston GI, et al. A phase 2 randomized, double-blind, placebo-controlled, proof-of-concept study of oral seletalisib in primary Sjögren's syndrome[J]. Rheumatology (Oxford), 2021, 60(3): 1364- 1375.

doi: 10.1093/rheumatology/keaa410
12
Hajiasgharzadeh K, Khabbazi A, Mokhtarzadeh A, et al. Choli-nergic anti-inflammatory pathway and connective tissue diseases[J]. Inflammopharmacology, 2021, 29(4): 975- 986.

doi: 10.1007/s10787-021-00812-z
13
Lee SM, Lee SW, Kang M, et al. FoxO1 as a regulator of aquaporin 5 expression in the salivary gland[J]. J Dent Res, 2021, 100(11): 1281- 1288.

doi: 10.1177/00220345211003490
14
Wang D, Zhao H, Li B, et al. Mechanism of cAMP-PKA signaling pathway mediated by Shaoyao Gancao decoction (芍药甘草汤) on regulation of aquaporin 5 and muscarinic receptor 3 levels in Sjögren's syndrome[J]. Chin J Integr Med, 2020, 26(7): 502- 509.

doi: 10.1007/s11655-020-3205-5
15
Chivasso C, Nesverova V, Järvå M, et al. Unraveling human AQP5-PIP molecular interaction and effect on AQP5 salivary glands localization in SS patients[J]. Cells, 2021, 10(8): 2108.

doi: 10.3390/cells10082108
16
Barrera MJ, Aguilera S, Castro I, et al. Dysfunctional mitochondria as critical players in the inflammation of autoimmune diseases: Potential role in Sjögren's syndrome[J]. Autoimmun Rev, 2021, 20(8): 102867.

doi: 10.1016/j.autrev.2021.102867
17
Yao Y, Ma JF, Chang C, et al. Immunobiology of T cells in Sjögren's syndrome[J]. Clin Rev Allergy Immunol, 2021, 60(1): 111- 131.

doi: 10.1007/s12016-020-08793-7
18
Wang D, Zhou M, Wang Y, et al. Suppression of high-mobility group box 1 ameliorates xerostomia in a Sjögren syndrome-triggered mouse model[J]. Can J Physiol Pharmacol, 2020, 98(6): 351- 356.

doi: 10.1139/cjpp-2019-0337
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