Comparison of anti-inflammatory, antibacterial and analgesic activities of formulated granules versus traditional decoction of Yinqiao Powder

doi:10.12122/j.issn.1673-4254.2025.05.13

Abstract

Abstract:

Objective To compare the anti-inflammatory, antibacterial and analgesic effects of Yinqiao Powder (YQS) formulated granules and decoction. Methods We first evaluated the anti-inflammatory effects of the two dosage forms of YQS in a LPS-induced RAW 264.7 cell model using RT-qPCR and Western blotting. We further constructed zebrafish models of inflammation by copper sulfate exposure, caudal fin transection, or LPS and Poly (I:C) microinjection, and evaluated anti-inflammatory effects of YQS granules and decoction by examining neutrophil aggregation and HE staining findings. In a mouse model of acute lung injury (ALI) induced by intratracheal LPS instillation, the effects of YQS gavage at 10, 15, and 20 g/kg on lung pathologies were evaluated by calculating lung wet-dry weight ratio and using HE staining, ELISA and Western blotting. The microbroth dilution method was used to evaluate the antibacterial effect of YQS. Mouse pain models established by hot plate and intraperitoneal injection of glacial acetic acid were used to evaluate the analgesic effects of YQS at 10, 15, and 20 g/kg. Results Both YQS granules and decoction significantly reduced TNF-α, IL-6, and IL-1β expressions and p-STAT3 (Tyr 705) phosphorylation level in LPS-induced RAW 264.7 cells, and obviously inhibited neutrophil aggregation in the zebrafish models. In ALI mice, YQS granules and decoction effectively ameliorated lung injury, lowered lung wet-dry weight ratio, and reduced p-STAT3 (Tyr 705) expression and TNF-α and IL-6 levels. YQS produced obvious antibacterial effect at the doses of 15.63 and 31.25 mg/mL, and significantly reduced body torsion and increased pain threshold in the mouse pain models. Conclusion The two dosage forms of TQS have similar anti-inflammatory, antibacterial and analgesic effects with only differences in their inhibitory effect on TNF-α, IL-6 and IL-1β mRNA expressions in LPS-induced RAW 264.7 cells.

Key words: Yinqiao Powder, formulated granules of traditional Chinese medicine, pharmacological equivalence

Zhuolin GUO, Zhiheng ZHANG, Xindeng GUO, Weiwei YANG, Zhiqing LIANG, Jinying OU, Huihui CAO, Zibin LU, Linzhong YU, Junshan LIU. Comparison of anti-inflammatory, antibacterial and analgesic activities of formulated granules versus traditional decoction of Yinqiao Powder[J]. Journal of Southern Medical University, 2025, 45(5): 1003-1012.

Figures/Tables 9

Tab.1 Primer sequences for RT-qPCR

Primer	Primer sequence (5'-3')	Primer length (nt)
Mouse-β-actin F	ATGGATGAGGAAATCGCTG	19
Mouse-β-actin R	ATGCCAACCATCACTCCCTG	20
Mouse-IL-6 F	ATGCCAACCATCACTCCCTG	20
Mouse-IL-6 R	TTTGATGTCGTTCACCAGGA	20
Mouse-TNF-α F	GCTGGATCTTCAAAGTCGGGTGTA	24
Mouse-TNF-α R	TGTGAGTCTCAGCACACTTCCATC	24
Mouse-IL-1β-F	TCGCAGCAGCACATCAACAAGAG	23
Mouse-IL-1β-R	TGCTCATGTCCTCATCCTGGAAGG	24

Fig.1 Effect of Yinqiao Powder granules (YQS KL) and decoction (YQS YP) on RAW 264.7 cells with LPS-induced inflammation. A, B: Cytotoxicity of YQS YP and KL in RAW 264.7 cells. C: Inhibitory effects of YQS YP and KL on expression of phosphorylated STAT3 in LPS-induced RAW 264.7 cells. D-F: Inhibitory effects of YQS YP and KL on mRNA expressions of TNF-α, IL-6 and IL-1β in LPS-induced RAW 264.7 cells (Mean±SD, n=3). ##P<0.01, ###P<0.001 vs control (C); **P<0.01,***P<0.001 vs Model (M) group.

Tab.2 Acute toxicity of Yinqiao Powder granules （YQS KL） and decoction （YQS YP） in zebrafish for 72 h

Group	Concentration (μg/mL)	Number of deaths	Mortality rate (%)
CON	-	0	0
YQS YP	100	0	0
	200	0	0
	400	0	0
	800	0	0
	1000	2	10
YQS KL	100	0	0
	200	0	0
	400	0	0
	800	0	0
	1000	1	5

Fig.2 Effects of YQS YP and KL on neutrophil aggregation in the yolk sac of a zebrafish model with LPS microinjection (Black arrow: Neutrophil) (HE staining, original magnification: ×200).

Fig.3 Effects of YQS YP and YQS KL on neutrophil aggregation in 4 zebrafish inflammation models. A, B: Zebrafish model of CuSO4-induced inflammation. C, D: Zebrafish model of inflammation with tail transection. E, F: Zebrafish model of inflammation induced by LPS microinjection. G, H: Zebrafish model of inflammation induced by poly (I: C) microinjection. ##P<0.01, ###P<0.001 vs control (C) group; *P<0.05,**P<0.01, ***P<0.001 vs Model (M) group.

Fig.4 Effects of YQS YP and YQS KL on lung tissue damage in mice with acute lung injury (HE staining, ×200).

Fig.5 Anti-inflammatory effects of YQS YP and YQS KL on acute lung injury mice. A: YQS YP and KL reduce wet-dry lung weight ratio of the mice. B-E: YQS YP and KL reduce TNF-α and IL-6 levels in mouse BALF and lung tissue. F: YQS YP and KL inhibit protein expression of p-STAT3 (Tyr 705) in mouse lung tissue. *P<0.05, **P<0.01,***P<0.001 vs LPS group; ###P<0.001 vs CON group.

Tab.3 Effects of YQS YP and YQS KL on pain threshold and number of twisted lifts in mice

Group	Dose (g/kg)	Pain threshold improvement rate(%)	Number of twists
CON	-	2.63±8.94	34.79±6.20
SLFSN	0.05	48.68±11.21***	16.15±3.71***
YQS YP	20	38.79±8.94***	17.67±6.44***
	15	21.75±8.24***	18.25±5.06***
	10	12.79±7.27	21.89±3.98***
YQS KL	20	45.53±10.71***	16.43±3.73***
	15	33.67±11.32***	18.80±4.31***
	10	8.90±9.34	23.50±2.61***

Tab.4 Minimum inhibitory concentration (MIC) of YQS YP and YQS KL against 3 bacteria (mg/mL)

Bacterial species	YQS YP (MIC)	YQS KL (MIC)
Escherichia coli	31.25	31.25
Staphylococcus aureus	15.63	15.63
Streptococcus pneumoniae	31.25	31.25

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