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ORIGINAL ARTICLE
Year : 2018  |  Volume : 131  |  Issue : 14  |  Page : 1724-1731

Effects of Dandelion Extract on the Proliferation of Rat Skeletal Muscle Cells and the Inhibition of a Lipopolysaccharide-Induced Inflammatory Reaction


1 Department of Radiology, Shengjing Hospital, China Medical University, Shenyang, Liaoning 110004, China
2 Department of Radiology, Shengjing Hospital, China Medical University, Shenyang, Liaoning 110004; Department of Radiology, The First Affiliated Hospital of University of South China, Hengyang, Hunan 421001, China

Correspondence Address:
Dr. Shi-Nong Pan
Department of Radiology, Shengjing Hospital, China Medical University, Shenyang, Liaoning 110004
China
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0366-6999.235878

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Background: Dandelion is commonly used in traditional Chinese medicine with several active compounds found in extracts. It has a variety of pharmacological effects, such as a reduction in swelling and inflammation, and detoxification. The mechanism by which dandelion extract inhibits the inflammatory response in skeletal muscle cells remains unknown; therefore, the aim of this study was to investigate the effects of dandelion extract root on the proliferation of skeletal muscle cells and the alleviation of lipopolysaccharide (LPS)-induced inflammatory response in vitro. Methods: Rat skeletal muscle cells were isolated from Sprague-Dawley rat and cultured in vitro which were cultured in basal medium, or medium containing LPS or dandelion extract. Cell counting kit-8 (CCK-8) was employed to measure cell proliferation; meanwhile, the optimal concentration of dandelion extract and treatment time were selected. Crystal violet staining was used to detect the proliferation of muscle cells. Western blotting analysis was used to detect the levels of inflammatory factors, myogenic factor, and p-AKT protein expression. Results: The optimal concentration and treatment time of dandelion extract for the following study were 5 mg/ml and 4 days, respectively. Dandelion extract was found to increase proliferation of rat skeletal muscle cells (t = 3.145, P < 0.05), with the highest effect observed at 5 mg/ml. LPS was found to decrease proliferation of skeletal muscle cells (t = −131.959, P < 0.001), and dandelion extract could against this affection (t = 19.466, P < 0.01). LPS could induce expression of inflammatory factors, including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α (IL-1β: t = 9.118, P < 0.01; IL-6: t = 4.346, P < 0.05; TNF-α: t = 15.806, P < 0.05), and dandelion extract was shown to reduce LPS-induced expression of IL-1β, IL-6 and TNF-α (IL-1β: t = −2.823, P < 0.05; IL-6: t = −3.348, P < 0.01; and TNF-α: t = −3.710, P < 0.01). Furthermore, LPS was also shown to decrease expression of myogenic factor, including myod1 and myogenin (MyoD1: t = 4.039, P < 0.05 and myogenin: t = 3.300, P < 0.01), but dandelion extract was shown to against this effect of LPS (MyoD1: t = −3.160, P < 0.05 and myogenin: t = −3.207, P < 0.01). And then, LPS was found to increase expression of p-AKT protein (p-AKT/AKT: t = 4.432, P < 0.05). Moreover, expression of p-AKT protein was found to decrease, with 5 mg/ml of dandelion extract (p-AKT/AKT: t = −3.618, P < 0.05). Conclusions: The findings indicate that dandelion extract plays an important role in skeletal muscle cells viability regulation, promote cells proliferation by increasing level of p-AKT protein expression, and reduce LPS-induced expression of inflammatory factors, inhibiting the inflammatory response of rat skeletal muscle cells.

 

 Abstract in Chinese

蒲公英提取物对大鼠骨骼肌细胞增殖和LPS诱导的炎症反应的抑制作用

摘要

背景: 蒲公英通常作为传统中医药,其提取物中含有几种活性化合物。蒲公英具有多种药理作用,如缓解肿胀、炎症,以及抗毒性作用等。但蒲公英提取物抑制骨骼肌细胞炎症反应的机制仍不清楚。因此,本研究的目的是探讨蒲公英根部提取物对骨骼肌细胞增殖的影响以及体外抑制LPS诱导的炎症反应。

方法: 从SD大鼠中分离大鼠骨骼肌细胞,并在体外进行普通培养基、含有脂多糖(LPS)或蒲公英提取物的培养基中培养。采用细胞计数试剂盒-8(CCK-8)对细胞增殖情况进行检测,同时找出蒲公英提取物的最佳浓度和最佳处理时间。结晶紫染色用于检测肌细胞的增殖情况。 Western blot检测并分析骨骼肌细胞中炎症因子,生肌因子和p-AKT蛋白的表达水平。

结果: 研究发现蒲公英提取物促进骨骼肌细胞增殖的最佳浓度和最佳处理时间分别为5mg / ml和4d。我们发现蒲公英提取物能够促进大鼠骨骼肌细胞增殖(t = 3.145,p <0.05),在5mg / ml时效果最佳。并且LPS可以抑制骨骼肌细胞增殖(t = -131.959,p <0.001),蒲公英提取物可以抵抗LPS的这种作用(t = 19.466,p <0.01)。 LPS可以诱导炎症因子IL-1β,IL-6和TNF-α的表达(IL-1β:t = 9.118,p <0.01; IL-6:t = 4.346,p <0.05; TNF-α: t=15.806, p<0.05),蒲公英提取物减少LPS诱导的IL-1β,IL-6和TNF-α的表达(IL-1β:t = -2.823,p <0.05; IL-6:t = -3.348,p <0.01; TNF -α:t = -3.710,p <0.01)。此外,LPS还显示降低生肌因子的表达,包括myod1和肌细胞生成素(myod1:t = 4.039,p <0.05;肌细胞生成素:t = 3.3,p <0.01),但蒲公英提取物显示出对抗LPS的这种作用 (myod1:t = -3.16,p <0.05;肌细胞生成素:t = -3.207,p <0.01)。我们进一步发现,LPS能够增加骨骼肌细胞p-AKT蛋白的表达(p-AKT / AKT:t = 4.432,p <0.05)。 此外,在LPS处理细胞后,用5mg / ml的蒲公英提取物发现p-AKT蛋白的表达减少(p-AKT / AKT:t = -3.618,p <0.05)。

结论: 研究结果表明,蒲公英提取物在骨骼肌细胞增殖调节中起重要作用,通过提高p-AKT蛋白表达水平促进细胞增殖,并减少LPS诱导的炎性因子表达,抑制大鼠骨骼肌细胞的炎症反应。



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