银翘散配方颗粒与传统汤剂在抗炎、抗菌及镇痛药效的差异比较

doi:10.12122/j.issn.1673-4254.2025.05.13

摘要/Abstract

摘要：

目的比较银翘散颗粒和传统汤剂的抗炎、抗菌、镇痛的药效差异。方法采用脂多糖（LPS）诱导巨噬细胞RAW 264.7构建炎症模型，并给予银翘散（200、400、800 μg/mL），利用实时荧光PCR、Western blotting评价药物抗炎效果；采用3 dpf斑马鱼分别构建硫酸铜、尾鳍横切、LPS和Poly （I∶C）显微注射斑马鱼炎症模型，并给予银翘散（200、400、800 μg/mL），利用中性粒细胞观察和HE染色等评价药物抗炎效果；采用LPS气管滴注构建急性肺损伤小鼠模型，并给予银翘散（10、15、20 g/kg），通过计算肺湿干质量比、HE染色、酶联免疫吸附法、Western blotting等实验评价药物抗炎效果。采用微量肉汤稀释法评价银翘散颗粒及其汤剂的抗菌效果。采用热板和冰醋酸腹腔注射构建小鼠疼痛模型，并给予银翘散（10、15、20 g/kg），通过计算小鼠热板痛阈值和醋酸扭体次数评价药物镇痛效果。结果与模型组相比，银翘散能降低细胞内炎症因子TNF-α、IL-6、IL-1β的水平（P<0.01）和p-STAT3（Tyr 705）蛋白表达水平；银翘散能抑制斑马鱼中性粒细胞的聚集（P<0.001）；银翘散可改善肺组织损伤，下调肺湿干质量比值、p-STAT3（Tyr 705）蛋白的表达、炎症因子TNF-α、IL-6的含量（P<0.05）；银翘散在15.63、31.25 mg/mL起到抑菌作用；银翘散能减少小鼠的扭体次数，提高痛阈值（P<0.05）。结论银翘散配方颗粒和传统汤剂均具有抗炎、抗菌、镇痛的作用，两者除在细胞水平上抑制TNF-α、IL-6、IL-1β的mRNA表达有差异外，在其他体内外多个实验评价中药效无明显差异。

关键词: 银翘散, 中药配方颗粒, 药物等效性

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

郭卓琳, 张智恒, 郭新邓, 杨苇巍, 梁芷晴, 区锦莹, 曹惠慧, 卢子滨, 余林中, 刘俊珊. 银翘散配方颗粒与传统汤剂在抗炎、抗菌及镇痛药效的差异比较[J]. 南方医科大学学报, 2025, 45(5): 1003-1012.

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.

图/表 9

表1 引物序列

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

图1 银翘散对LPS诱导的RAW 264.7细胞炎症模型的影响

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.

表2 银翘散作用于斑马鱼72 h的急性毒性结果

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

图2 银翘散颗粒和汤剂对显微注射LPS斑马鱼模型中性粒细胞在卵黄囊聚集的影响

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).

图3 银翘散颗粒和汤剂对4种斑马鱼炎症模型中性粒细胞的影响

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.

图4 银翘散颗粒和汤剂对急性肺损伤小鼠肺组织损伤的影响

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

图5 银翘散颗粒和汤剂对急性肺损伤小鼠的影响

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.

表3 银翘散颗粒和汤剂对小鼠痛阈值和扭体次数的影响

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***

表4 银翘散颗粒和汤剂对3种细菌的最小抑菌浓度

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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