Anti-inflammatory activities of Guang-Pheretima extract in lipopolysaccharide-stimulated RAW 264.7 murine macrophages

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.

Antioxidant and Anti-Inflammatory Activities of the Crude Extracts of Moringa oleifera from Kenya and Their Correlations with Flavonoids

“…The murine macrophage RAW264.7 cells were obtained from the American Type Culture Collection (ATCC), and cultured at 37 °C in the DMEM medium supplemented with the 10% FBS and 1% penicillin-streptomycin in a humidified incubator containing 5% CO 2 . The nitrite production was measured according to the Griess reagent protocol [33]. Briefly, the cells were transferred into the 96-well plate (4–6 × 10 4 cells in each well) and incubated for 24 h. Then, those cells were cultured with serial dilution (11.11, 33.33, 100 μg/mL) of leaves and seeds extracts of M. oleifera for 2 h. Next, those cells were added with the 10 ng/mL of LPS for another 24 h. Then, 100 μL of the culture medium were mixed with the equal volume of the Griess reagent, and after 15 min of reaction, the mixture absorbance was read at 540 nm with a microplate reader.…”

Antioxidant, Anti-inflammatory Activities and Polyphenol Profile of Rhamnus prinoides

“…The cells were maintained at 37 °C in DMEM medium supplemented with the 10% FBS, 1% penicillin–streptomycin with 5% CO 2 in a humidified incubator. The nitrite concentration, an indicator of the NO synthesis, was measured based on the previously described Griess reaction method [ 42 ]. Briefly, RAW 264.7 cells were transferred into the 96-well plate with a density of 5 × 10 4 cells/well and kept for 24 h. These cells were incubated with various concentrations of the SPRE solution for 2 h and then followed by the incubation with 10 ng/mL of LPS for another 24 h. Meanwhile, 10.0 μg/mL of aspirin was used as the positive control.…”

An Integrated Analysis of Network Pharmacology and Experimental Validation to Reveal the Mechanism of Chinese Medicine Formula Naotaifang in Treating Cerebral Ischemia-Reperfusion Injury

“…49,50 Previous reports have shown that Pheretima could exert anti-inflammatory effects by reducing the production of inflammatory mediators such as NO, PGE2, TNF-α and various inflammatory cytokines. 51 There is a research revealed that Pheretima may have anti-coagulant and anti-thrombotic functions through regulating DPf3, which is a group of antithrombotic functional proteins purified from Pheretima. 52 Moreover, Bombyx Batryticatus has been reported to exhibit pharmacological activity on the nervous system, such as anticonvulsant and antiepileptic effects, hypnotic effects, and neurotrophic effects and so on.…”