Abstract
BackgroundGuang-Pheretima, which is originated from Pheretima aspergillum, has been documented in academic Chinese herbal studies for nearly 2000 years for its prominent treating effects of various inflammatory diseases such as asthma, cough and fever. However, the anti-inflammatory activity and mechanism of Guang-Pheretima has been rarely reported. Hence, we investigated the inhibitory effect and the underlying mechanism of Guang-Pheretima aqueous extracts on inflammatory response in RAW 264.7 cells.MethodRAW 264.7 macrophages were pretreated with various concentrations of Guang-Pheretima decoction (GPD) or protein-free Guang-Pheretima decoction (PF-GPD) and subsequently stimulated with lipopolysaccharide (LPS) to trigger the inflammatory response. Productions of nitric oxide (NO) were determined by Griess reaction, and prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6 were measured by enzyme-linked immunosorbent assays (ELISA). The protein expressions and messenger ribonucleic acid (mRNA) amounts of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were analyzed by Western Blot and Real-Time polymerase chain reaction (PCR), respectively. Finally, the translocation of nuclear factor (NF)-κB was observed by Western Blot.ResultsGPD of the experimental concentrations showed no anti-inflammatory activity. In contrast, PF-GPD at concentrations of 40–320 μg/mL significantly inhibited NF-κB activation and reduced the production of inflammatory mediators, such as NO, PGE2, TNF-α, as well as the related key synthases including iNOS and COX-2. Moreover, PF-GPD markedly suppressed the release of inflammatory cytokines, such as IL-1β and IL-6.ConclusionThese results demonstrate the excellent anti-inflammatory properties of PF-GPD, and suggest that Guang-Pheretima may be used to treat and prevent certain inflammatory diseases.
Highlights
Guang-Pheretima, which is originated from Pheretima aspergillum, has been documented in academic Chinese herbal studies for nearly 2000 years for its prominent treating effects of various inflammatory diseases such as asthma, cough and fever
Effect of Guang-Pheretima decoction (GPD) and protein-free Guang-Pheretima decoction (PF-GPD) on nitric oxide (NO) synthesis To evaluate the effect of GPD and PF-GPD on the synthesis of the inflammatory mediator NO, we performed a Griess test to measure the concentration of NaNO2 in the conditioned medium of LPS-stimulated RAW 264.7 cells
Effect of GPD and PF-GPD on prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and proinflammatory cytokines PGE2 and TNF-α are important mediators of inflammation, while IL-1β and IL-6 are pro-inflammatory cytokines that are secreted in the early stages of inflammation
Summary
Guang-Pheretima, which is originated from Pheretima aspergillum, has been documented in academic Chinese herbal studies for nearly 2000 years for its prominent treating effects of various inflammatory diseases such as asthma, cough and fever. We investigated the inhibitory effect and the underlying mechanism of Guang-Pheretima aqueous extracts on inflammatory response in RAW 264.7 cells. Inflammation, mainly manifesting as redness, swelling, fever and pain, is a bodily defense mechanism in response to injury, infection and other stimuli, and plays a vital role in defending against diseases and pathogen invasion. During inflammation, activated macrophages secrete excessive inflammatory mediators, such as nitric oxide (NO) and prostaglandins (PGs), as well as pro-inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) [1]. During the course of inflammation, NO free radicals that are produced by inducible nitric oxide synthase (iNOS) play an important role in the innate immune response to infection [4]. The stimulated macrophages produce large amounts of inflammatory cytokines, including TNF-α, IL-1β
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