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[Effect of Jingfang Granules on carrageenan-induced tail thrombosis in mice based on ERK/p38 MAPK signaling pathway]
Ji-Dong Zhou1, Hong-Hua Li1, Xiang-Zi Li1
1New Drug Pharmacology Center of Lunan Pharmaceutical Group Co., Ltd. Linyi 276006, China State Key Laboratory of Generic Technology of Traditional Chinese Medicine Linyi 276006, China Linyi Key Laboratory of Immunopharmacology and Toxicology of Natural Drugs Linyi 276006, China.
Abstract:
The present study explored the anti-inflammatory and anti-thrombotic mechanism of Jingfang Granules on tail thrombosis induced by carrageenan in mice. Thirty-two male ICR mice were randomly divided into a control group, a model group, a Jingfang Granules group, and a positive drug(aspirin) group, with eight mice in each group. The thrombosis model was induced by intraperitoneal injection of carrageenan(45 mg·kg~(-1)) combined with low-temperature stimulation, and the mice were treated with drugs for 7 days before modeling. Twenty-four hours after modeling, blood was detected for four blood coagulation indices in each group. The enzyme-linked immunosorbent assay(ELISA) was used to detect the activity of plasma interleukin-6(IL-6), interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and other inflammatory factors. The tails of mice in each group were cut off to observe tail lesions and measure the length of the thrombus. The protein expression and phosphorylation level of extracellular signal-regulated kinase 1/2(ERK1/2) and p38 mitogen-activated protein kinase(p38 MAPK) in spleen tissues were detected by Western blot. The results showed that dark red thrombus appeared in the tails of mice in each group. The length of the black part accounted for about 40% of the total tail in the model group. Additionally, the model group showed prolonged prothrombin time(PT), increased fibrinogen(FIB) content, and shortened activated partial thromboplastin time(APTT). Compared with the model group, the groups with drug intervention displayed shortened black parts in the tail and improved four blood coagulation indices(P<0.05). As revealed by ELISA, the expression levels of TNF-α, IL-1β, and IL-6 in the mouse plasma were significantly up-regulated in the model group, and those in the groups with drug intervention were reduced as compared with the model group(P<0.05). As demonstrated by Western blot, the protein expression and phosphorylation levels of ERK1/2 and p38 MAPK in the spleen tissues were significantly elevated in the model group, while those in the Jingfang Granules group were down-regulated as compared with the model group with a significant difference. Jingfang Granules can inhibit tail thrombosis of mice caused by carrageenan presumedly by inhibiting the activation of ERK1/2 and p38 MAPK signaling pathways.
Insights
Jingfang Granules effectively reduced tail thrombosis in mice by improving blood coagulation and decreasing inflammatory markers. The treatment inhibited the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and p38 mitogen-activated protein kinase (p38 MAPK) signaling pathways.
Area of Science:
- Pharmacology
- Immunology
- Biochemistry
Background:
- Thrombosis is a significant health concern, often associated with inflammation.
- Traditional Chinese Medicine, like Jingfang Granules, is explored for its therapeutic potential.
- Understanding the molecular mechanisms behind anti-thrombotic agents is crucial for drug development.
Purpose of the Study:
- To investigate the anti-inflammatory and anti-thrombotic effects of Jingfang Granules.
- To elucidate the underlying molecular mechanisms, focusing on inflammatory markers and signaling pathways.
- To evaluate the efficacy of Jingfang Granules in a mouse model of carrageenan-induced tail thrombosis.
Main Methods:
- A carrageenan-induced tail thrombosis model was established in ICR mice.
- Mice were treated with Jingfang Granules or aspirin (positive control) prior to thrombosis induction.
- Blood coagulation indices, plasma inflammatory cytokines (IL-6, IL-1β, TNF-α) via ELISA, and spleen protein expression/phosphorylation of ERK1/2 and p38 MAPK via Western blot were analyzed.
Main Results:
- Jingfang Granules significantly reduced thrombus length and improved blood coagulation parameters (PT, FIB, APTT) compared to the model group.
- The treatment markedly decreased elevated levels of plasma inflammatory cytokines (TNF-α, IL-1β, IL-6).
- Jingfang Granules notably downregulated the protein expression and phosphorylation of ERK1/2 and p38 MAPK in spleen tissues.
Conclusions:
- Jingfang Granules exhibit significant anti-inflammatory and anti-thrombotic properties.
- The therapeutic effects are likely mediated by the inhibition of ERK1/2 and p38 MAPK signaling pathways.
- These findings support the potential of Jingfang Granules as a treatment for thrombotic and inflammatory conditions.
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