Korean J Pain.  2020 Oct;33(4):335-343. 10.3344/kjp.2020.33.4.335.

Effect of Zhongyi paste on inflammatory pain in mice by regulation of the extracellular regulated protein kinases 1/2– cyclooxygenase-2–prostaglandin E₂ pathway

• Xiao A
• Wu C
• Kuang L
• Lu W
• Zhao X
• Kuang Z
• Hao N

• KMID: 2507139
• DOI: http://doi.org/10.3344/kjp.2020.33.4.335

Abstract

Background
Zhongyi paste is a traditional Chinese medicine herbal paste that is externally applied to reduce inflammation and relieve pain.
Methods
An acute foot swelling inflammation model in C57BL/6J mice was established by carrageenan-induced pathogenesis. Zhongyi paste raised the pain threshold and also reduced the degree of swelling in mice with carrageenan-induced foot swelling.
Results
Analysis indicated that serum tumor necrosis factor-alpha, interleukin-1 beta, and prostaglandin E₂ (PGE₂) cytokine levels and PGE₂levels in the paw tissue of the mice were decreased by Zhongyi paste treatment. The quantitative polymerase chain reaction and western blot results showed that Zhongyi paste downregulated the mRNA and protein expression of extracellular signal-regulated kinase 1/2 (ERK1/2), and cyclooxygenase-2 (COX-2), and also downregulated the mRNA expression of PGE₂ . At the same time, the Zhongyi paste exerted a stronger effect as an external drug than that of indomethacin, which is an oral drug, and voltaren, which is an externally applied drug.
Conclusions
Our results indicated that Zhongyi paste is a very effective drug to reduce inflammatory swelling of the foot, and its mechanism of action is related to regulation of the ERK1/2–COX-2–PGE₂ pathway.

Keyword

Biochemistry; Cyclooxygenase 2; Herbal Medicine; Inflammation; Medicine; Chinese Traditional; Mitogen-Activated Protein Kinase 3; Pain; RNA; messenger

Figure

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  Fig. 1 The paw volume meter in this study. Paw volume meter: KW-7C, Nanjing Calvin Biotechnology Co., Ltd, Nanjing, Jiangsu, China.

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  Fig. 2 Effects of Zhongyi paste on pain threshold in normal mice. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste: paw swollen mice evenly applied with Zhongyi paste.

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  Fig. 3 Effects of Zhongyi paste on pain threshold in mice with carrageenan-induced foot swelling. Values presented are the mean ± standard deviation. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste: paw swollen mice evenly applied with Zhongyi paste. a-eMean values with different letters in the bar are significantly different (P < 0.05) according to Duncan’s new multiple range test. a < 0.05 vs. model group, b < 0.05 vs. model group, c < 0.05 vs. model group, d < 0.05 vs. model group, e < 0.05 vs. normal group.

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  Fig. 4 Effects of Zhongyi paste on swelling degree of foot in carrageenan-induced foot swelling mice. Values presented are the mean ± standard deviation. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste: paw swollen mice evenly applied with Zhongyi paste. a-dMean values with different letters in the bar are significantly different (P < 0.05) according to Duncan’s new multiple range test. a < 0.05 vs. normal group, b < 0.05 vs. model group, c < 0.05 vs. model group, d < 0.05 vs. model group.

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  Fig. 5 Effects of Zhongyi paste on paw tissue prostaglandin E2 (PGE2) levels in carrageenan-induced foot swelling mice. Values presented are the mean ± standard deviation. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste: paw swollen mice evenly applied with Zhongyi paste. a-eMean values with different letters in the bar are significantly different (P < 0.05) according to Duncan’s new multiple range test. a < 0.05 vs. normal group, b < 0.05 vs. model group, c < 0.05 vs. model group, d < 0.05 vs. model group, e < 0.05 vs. model group.

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  Fig. 6 Effects of Zhongyi paste on paw tissue extracellular signal-regulated kinase 1/2 (ERK1/2), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2) mRNA expression in carrageenan-induced foot swelling mice. Values presented are the mean ± standard deviation. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste group: paw swollen mice evenly applied with Zhongyi paste. a-eMean values with different letters in the bar are significantly different (P < 0.05) according to Duncan’s new multiple range test. a < 0.05 vs. normal group, b < 0.05 vs. model group, c < 0.05 vs. model group, d < 0.05 vs. model group, e < 0.05 vs. model group.

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  Fig. 7 Effects of Zhongyi paste on paw tissue extracellular signal-regulated kinase 1/2 (ERK1/2) and cyclooxygenase-2 (COX-2) protein expression in carrageenan-induced foot swelling mice. Values presented are the mean ± standard deviation. Indomethacin: paw swollen mice treated with 10 mg/kg body weight indomethacin by gavage, Voltaren: paw swollen mice evenly applied with voltaren, Zhongyi paste: paw swollen mice evenly applied with Zhongyi paste. a-eMean values with different letters in the bar are significantly different (P < 0.05) according to Duncan’s new multiple range test. a < 0.05 vs. normal group, b < 0.05 vs. model group, c < 0.05 vs. model group, d < 0.05 vs. model group, e < 0.05 vs. model group.

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