Study on the effect of ZiYin RunZao (ZYRZ) pill on the inflammation and function of submandibular gland in mice with Sjögren's syndrome

doi:10.12092/j.issn.1009-2501.2026.02.007

Abstract

Abstract:

AIM: To investigate the effect of ZiYin RunZao (ZYRZ) pill on the inflammation and function of submandibular gland in mice modeled with Sj?gren's syndrome. METHODS: Forty 8-week-old NOD/Ltj mice were randomly divided into five groups: model group, western medicine group, and the high, middle and low dosage groups of ZYRZ, with eight mice in each group. Another 8 ICR mice were blank group. The intervention was started after 1 week of acclimatization feeding. In the high, medium and low dose groups, 1.78, 0.89 and 0.45 mg/kg of ZYRZ pills were administered by gavage, respectively; in the western medicine group, 60 mg/kg of hydroxychloroquine was administered; and in the blank group and the model group, equal amounts of sterile distilled water were administered by gavage once a day for eight consecutive weeks. The body weight and water intake of the mice were monitored at weeks 0, 2, 4, 6 and 8. At the end of the intervention, mice submandibular gland and serum were taken, and the histopathology of submandibular gland was observed by HE staining; the levels of inflammatory cytokines IFN-γ, IL-10, and TNF-α were detected in serum by ELISA; the levels of AQP5 and M3R mRNA were detected in submandibular gland by RT-qPCR; the levels of AQP5 and M3R mRNA in submandibular gland were detected in submandibular gland by Western blot; and the levels of AQP5 and M3R mRNA in submandibular gland were detected in submandibular gland by RT-qPCR. AQP5, M3R protein expression; immunohistochemistry to detect the level of AQP5 protein expression in the submandibular gland tissue of mice. RESULTS: Compared with the normal group, mice in the model group showed no significant changes in body weight, significantly increased water intake (P<0.05), significantly higher expression of IFN-γ and TNF-α in serum of mice (P<0.05), and significantly lower expression of IL-10 (P<0.05); significantly lower expression of mRNA and protein of AQP5 and M3R (P<0.05); compared with the model group, the western blot and the ZYRZ Pill in high school and the western blot were detected in the submaxillary gland tissues of mice; immunohistochemical method was used to detect the expression level of AQP5 in the submaxillary gland tissues. Compared with the model group, mice in the western medicine group and the high school dose group of ZYRZ consumed significantly less water (P<0.05), the expression of IFN-γ and TNF-α in serum was significantly reduced (P<0.05), the expression of IL-10 was significantly elevated (P<0.05), and the expression of AQP5, M3R mRNA, and protein was significantly elevated (P<0.05). CONCLUSION: ZYRZ pill can reduce the inflammatory reaction of submandibular gland in mice with Sj?gren's syndrome model and improve the secretion function of submandibular gland.

Key words: ZiYin RunZao pill, Sjgren's syndrome, submaxillary gland, inflammation, secretory function

CLC Number:

Wenxia QI, Gang WANG, Yanfeng YAN, Jiexiang TIAN, Hailong LIU, Yong WEI, Jia WANG. Study on the effect of ZiYin RunZao (ZYRZ) pill on the inflammation and function of submandibular gland in mice with Sjögren's syndrome[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(2): 196-203.

Figures/Tables 8

Table 1 qPCR primers

Gene	Primer type	Primer sequence
AQP5	Forward	CATGAACCCAGCCCGATCTT
AQP5	Reverse	CTCAGCGAGGAGGGGAAAAG
M3R	Forward	ACGAACAACAAAACGAGCCG
M3R	Reverse	AGATGACCGTTTCGTGCCTT

Table 2 Comparison of water intake among different groups of mice ($ \overline{x} $±s, n=8)

Group	Dose (g/kg)	Daily water intake (g)
Group	Dose (g/kg)	0 week	2 weeks	4 weeks	6 weeks	8 weeks
Normal	-	3.82±0.08	3.98±0.11	4.16±0.09	4.27±0.12	4.44±0.07
Model	-	4.11±0.08^b	4.65±0.20^b	5.18±0.15^b	5.41±0.34^b	6.20±0.18^b
Hydroxychloroquinb sulfatb	60 mg/kg	4.25±0.09^b	4.60±0.13^b	4.77±0.15^be	4.75±0.15^be	4.69±0.13^be
Ziyinrunzao pill low	0.45	4.20±0.11^b	4.50±0.11^b	4.81±0.09^be	4.86±0.13^be	4.97±0.28^be
Ziyinrunzao pill medium	0.89	4.28±0.09^b	4.43±0.09^b	4.70±0.24^be	4.68±0.19^be	4.64±0.17^be
Ziyinrunzao pill high	1.78	4.26±0.09^b	4.50±0.15^b	4.77±0.08^be	4.73±0.07^be	4.68±0.07^be

Fig.1 Pathological changes of SMG in different groups of mice (HE staining, 200x) NC: Normal group; MC: Model group; HCQ: Hydroxychloroquinb sulfatb group; ZL: Ziyinrunzao pill low group; ZM: Ziyinrunzao pill medium group; ZH: Ziyinrunzao pill high group.

Table 3 Comparison of IFN-γ, IL-10, and TNF-α levels in serum of different groups of mice ($ \overline{x} $±s, n=8)

Group	Dose (g/kg)	IFN-γ (ng/L)	IL-10 (pg/mL)	TNF-α (ng/L)
Normal	-	169.75±26.52	326.27±31.78	175.01±20.04
Model	-	365.43±34.30^b	205.98±30.51^b	381.01±37.32^b
Hydroxychloroquinb sulfatb	60 mg/kg	289.5±24.32^be	243.43±36.44^be	287.68±29.90^be
Ziyinrunzao pill low	0.45	293.83±32.59^be	235.09±26.00^be	298.74±27.61^be
Ziyinrunzao pill medium	0.89	260.80±35.17^be	285.39±45.16^be	281.61±18.16^be
Ziyinrunzao pill high	1.78	217.92±20.53^be	297.84±26.99^be	249.25±27.98^be

Table 4 Comparison of AQP5 and M3R mRNA in SMG tissues of different groups of mice ($ \overline{x} $±s, n=8)

Group	Dose (g/kg)	AQP5	M3R
Normal	-	1.31±0.09	1.54±0.11
Model	-	0.62±0.06^b	0.54±0.07^b
Hydroxychloroquinb sulfatb	60 mg/kg	0.89±0.02^be	1.00±0.11^be
Ziyinrunzao pill low	0.45	1.02±0.04^be	0.73±0.09^be
Ziyinrunzao pill medium	0.89	0.91±0.06^be	1.08±0.04^be
Ziyinrunzao pill high	1.78	0.95±0.03^be	1.18±0.06^be

Fig.2 Comparison of AQP5 and M3R mRNA in SMG tissues of different groups of mice ($ \overline{x} $±s, n=8) NC: Normal group; MC: Model group; HCQ: Hydroxychloroquinb sulfatb group; ZL: Ziyinrunzao pill low group; ZM: Ziyinrunzao pill medium group; ZH: Ziyinrunzao pill high group. bP<0.05, compared with normal group; eP<0.05, compared with model group.

Fig.3 Comparison of AQP5 and M3R protein levels in SMG tissues of different groups of mice ($ \overline{x} $±s, n=8) NC: Normal group; MC: Model group; HCQ: Hydroxychloroquinb sulfatb group; ZL: Ziyinrunzao pill low group; ZM: Ziyinrunzao pill medium group; ZH: Ziyinrunzao pill high group. bP<0.05, compared with normal group; eP<0.05, compared with model group.

Fig.4 Comparison of AQP5 protein levels in SMG tissues of different groups of mice (IHC, $ \overline{x} $±s, n=8) NC: Normal group; MC: Model group; HCQ: Hydroxychloroquinb sulfatb group; ZL: Ziyinrunzao pill low group; ZM: Ziyinrunzao pill medium group; ZH: Ziyinrunzao pill high group. bP<0.05, compared with normal group; eP<0.05, compared with model group.

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