Mechanistic study of paeoniflorin in ameliorating lacrimal gland inflammatory injury associated with Sjögren's syndrome

doi:10.12092/j.issn.1009-2501.2026.09.007

Abstract

Abstract:

AIM: To preliminarily predict the potential action direction of paeoniflorin in alleviating lacrimal gland inflammatory injury associated with Sj?gren's syndrome (SS) based on target screening and pathway analysis, and to provide preliminary validation through molecular docking and animal experiments. METHODS: Candidate targets of paeoniflorin and SS were screened from public databases and subjected to pathway enrichment analysis to identify inflammation-related signaling directions. Molecular docking was then performed to evaluate the binding characteristics between paeoniflorin and key inflammatory proteins. An SS-related dry eye model was established in male NOD/ShiLtJ mice, which were treated with different doses of paeoniflorin or hydroxychloroquine for four weeks. Tear secretion was measured to assess lacrimal gland function. Histopathological changes were evaluated by hematoxylin–eosin (HE) staining, and the expression levels of inflammatory mediators and signaling proteins in lacrimal glands were determined using RT-qPCR and western blot. RESULTS: Pathway analysis indicated that inflammasome-related signaling, particularly the NOD-like receptor pathway, may be involved in the action of paeoniflorin. Molecular docking showed that paeoniflorin exhibited favorable binding affinity to NLRP3 and Caspase-1. Animal experiments indicated that paeoniflorin treatment improved tear secretion, alleviated lacrimal gland inflammatory infiltration and tissue injury in SS mice, and was accompanied by reduced expression levels of IL-6, IL-18, IL-1β, TNF-α, NLRP3, and Caspase-1. CONCLUSION: This study provides preliminary evidence that paeoniflorin may ameliorate SS-related lacrimal gland inflammatory injury, possibly through modulation of inflammatory responses and NLRP3/Caspase-1–related processes.

Key words: paeoniflorin, Sj?gren's syndrome–related dry eye, lacrimal gland inflammatory injury, NLRP3/Caspase-1 signaling pathway

CLC Number:

Xuemei HAN, Hua ZHOU, Chunzi LIU. Mechanistic study of paeoniflorin in ameliorating lacrimal gland inflammatory injury associated with Sjögren's syndrome[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(9): 1214-1220.

Figures/Tables 7

Table 1 Primer sequences

Gene	Forward Primer	Reverse Primer
Mouse IL-6	CACTTCACAAGTCGGAGGCT	GCCACTCCTTCTGTGACTCC
Mouse IL-1β	TTGAAGTTGACGGACCCCAA	TGTCCTGACCACTGTTGTTTC
Mouse IL-18	TCAGACAACTTTGGCCGACT	TCAGTCTGGTCTGGGGTTCA
Mouse TNF-α	AGGCACTCCCCCAAAAGATG	CCACTTGGTGGTTTGTGAGTG
Mouse GAPDH	GGGTCCCAGCTTAGGTTCAT	CTCGTGGTTCACACCCATCA

Fig.1 Candidate target screening and key inflammatory pathway analysis of paeoniflorin in Sj?gren's syndrome A: venn diagram of overlapping targets between paeoniflorin and Sjögren's syndrome; B: protein-protein interaction network of the intersecting targets; C: KEGG enrichment bubble plot showing inflammation-related pathways.

Fig.2 Molecular docking of paeoniflorin with core target proteins

Fig.3 Effects of paeoniflorin on tear secretion in SS mice (x?±s, n=3) Tear secretion was measured at weeks 0, 2, and 4 in each group. cP<0.01, compared with control group; fP<0.01, compared with SS model group.

Fig.4 Effects of paeoniflorin on lacrimal gland histopathology in SS mice (x?±s, n=3) Representative HE-stained images of lacrimal gland tissues and quantitative analysis of inflammatory infiltration scores were shown. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with SS model group.

Fig.5 Effects of paeoniflorin on inflammatory cytokine mRNA expression in lacrimal glands of SS mice (x?±s, n=3) Relative mRNA expression levels of IL-6, IL-1β, IL-18, and TNF-α in lacrimal gland tissues were shown. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with SS model group.

Fig.6 Effects of paeoniflorin on inflammation-related protein expression in lacrimal glands of SS mice (x?±s, n=3) Western blot results and quantitative analysis of NLRP3, Caspase-1, and GSDMD protein expression in lacrimal gland tissues are shown. cP<0.01, compared with control group; eP<0.05, fP<0.01, compared with SS model group.

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