Introduction: Chronic inflammation is implicated in a multitude of diseases, including arthritis, neurodegeneration, autoimmune myositis, type 2 diabetes, rheumatic disorders, spondylitis, and cancer. Therefore, strategies to explore potent anti-inflammatory regimens are pivotal from a human-health perspective. Medicinal plants represent a vast unexplored treasure trove of therapeutically active constituents with diverse pharmacological activities, including anti-inflammatory properties. Herein, we evaluated Cousinia thomsonii, an edible medicinal herb, for its anti-inflammatory/immunomodulatory properties.
Methods: Soxhlet extraction was used to obtain different solvent extracts (hexane, ethyl acetate, ethanol, methanol, and aqueous extract) in increasing order of polarity. In vitro anti-inflammatory assays were performed to investigate the effects of extracts on protein denaturation, proteinase activity, nitric oxide surge, and erythrocyte-membrane stabilization. The most effective extracts, ie, ethyl acetate (CTEA) and ethanol (CTE) extracts (150– 200 g) were selected for further in vivo analysis using albino Wistar rats. Wistar rats received varying concentrations of CTEA and CTE (25, 50, and 100 mg/kg) for 3 weeks, followed by a single subplantar injection of lipopolysaccharide. Dexamethasone served as positive control. Blood was obtained from the retro-orbital plexus and serum separated for estimation of proinflammatory cytokines (IL6, IL1β, IFNγ and TNFα). Western blotting was performed to study expression patterns of crucial proteins implicated in the NFκB pathway, ie, NFκB p65, NFκB1 p50, and NFκB2 p52. Histopathological examination was done and gas chromatography–mass spectrometry (GC-MS) carried out to reveal the identity of compounds responsible for ameliorating effects of C. thomsonii.
Results: Among five tested extracts, CTEA and CTE showed marked inhibition of protein denaturation, proteinase activity, nitric oxide surge and erythrocyte-membrane hemolysis at 600 μg/mL (P< 0.001). Both these extracts showed no toxic effects up to a dose of 2,500 mg/kg. Extracts exhibited concentration-dependent reductions in expression of IL6, IL1β, IFNγ, TNFα, NFκB-p65, NFκB1, and NFκB2 (P< 0.05). Healing effects of extracts were evident from histopathological investigation. GC-MS analysis revealed the presence of important anti-inflammatory compounds, notably stigmast-5-en-3-ol, oleate, dotriacontane, ascorbic acid, n-hexadecanoic acid, and α-tocopherol, in C. thomsonii.
Conclusion: C. thomsonii possesses significant anti-inflammatory/immunomodulatory potential by virtue of modifying levels of proinflammatory cytokines/markers and NFκB proteins.

Keywords: chronic inflammation, interleukin, cytokine, immunoresponse, nitric oxide, compounds


中文翻译：

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活性 Cousinia thomsonii 提取物调节脂多糖诱导的白化 Wistar 大鼠模型中关键促炎介质/细胞因子和 NFκB 级联的表达


简介：慢性炎症与多种疾病有关，包括关节炎、神经退行性疾病、自身免疫性肌炎、2 型糖尿病、风湿性疾病、脊柱炎和癌症。因此，从人类健康的角度来看，探索有效的抗炎方案的策略至关重要。药用植物代表着一个巨大的未开发的治疗活性成分宝库，具有多种药理活性，包括抗炎特性。在此，我们评估了Cousinia thomsonii （一种食用药草）的抗炎/免疫调节特性。

方法：采用索氏提取，按极性递增顺序获得不同溶剂提取物（己烷、乙酸乙酯、乙醇、甲醇和水提取物）。进行体外抗炎测定，以研究提取物对蛋白质变性、蛋白酶活性、一氧化氮激增和红细胞膜稳定的影响。选择最有效的提取物，即乙酸乙酯 (CTEA) 和乙醇 (CTE) 提取物（150-200 g），使用白化 Wistar 大鼠进行进一步的体内分析。 Wistar 大鼠接受不同浓度的 CTEA 和 CTE（25、50 和 100 mg/kg）3 周，然后单次足底注射脂多糖。地塞米松作为阳性对照。从眼眶后丛获取血液并分离血清以评估促炎细胞因子（IL6、IL1β、IFNγ和TNFα）。进行蛋白质印迹来研究 NFκB 通路中涉及的关键蛋白（即 NFκB p65、NFκB1 p50 和 NFκB2 p52）的表达模式。我们进行了组织病理学检查和气相色谱-质谱分析 (GC-MS)，以揭示负责C. thomsonii 改善作用的化合物的身份。

结果：在五种测试提取物中，CTEA 和 CTE 在 600 μg/mL 浓度下对蛋白质变性、蛋白酶活性、一氧化氮激增和红细胞膜溶血具有显着抑制作用 ( P < 0.001)。这两种提取物在剂量高达 2,500 毫克/公斤时均未显示出毒性作用。提取物表现出 IL6、IL1β、IFNγ、TNFα、NFκB-p65、NFκB1 和 NFκB2 表达浓度依赖性降低 ( P < 0.05)。组织病理学研究表明提取物的治疗作用是显而易见的。 GC-MS 分析表明，C. thomsonii中存在重要的抗炎化合物，特别是 stigmast-5-en-3-ol、油酸盐、三十烷、抗坏血酸、正十六烷酸和 α-生育酚。

结论：托氏梭菌通过改变促炎细胞因子/标记物和 NFκB 蛋白的水平而具有显着的抗炎/免疫调节潜力。


关键词：慢性炎症、白细胞介素、细胞因子、免疫反应、一氧化氮、化合物