ABSTRACT
Objective:To explore the effect of Ermiaosan（EMS） on the polarization of M1 by lipopolysaccharide（LPS）+interferon(IFN)-γ and M2 induced by interleukin(IL)-4+IL-13 in rat bone marrow-derived macrophages. Method:Macrophages from rat bone marrow were extracted in vitro, stimulated by macrophage colony stimulating factor(M-CSF), induced to macrophages (marked by F4/80), stimulated by LPS+IFN-γ and induced to polarize to M1，while stimulated by IL-4+IL-13 and induced to polarize to M2. After adding different concentrations of EMS (0.2，0.4，0.8 g·L-1), the phenotypes of M1 and M2 were detected by immunofluorescence, and the effect of EMS on M1(marked by CD68 and iNOS)/M2(marked by CD206 and Arginase) polarization of macrophages from rat bone marrow was detected. Result:Compared with control group, LPS + IFN-γ could increase the polarization of M1 (P<0.01)，while IL-4+IL-13 could increase the polarization of M2 (P<0.01); compared with LPS+IFN-γ/IL-4+IL-13 group, EMS (0.2，0.4，0.8 g·L-1) could inhibit the polarization of M1 induced by LPS+IFN -γ for 24 hours (P<0.05), but had no significant effect on polarization of M2 induced by IL-4+IL-13. Conclusion:EMS can inhibit M1 polarization induced by LPS+IFN - γ, but has no effect on M2 polarization induced by IL-4+IL-13.
ABSTRACT
Objective::To study the effects of Ermiaosan on migration, adhesion and invasion of human fibroblast-liked synovial cells(FLS) and explore its mechanism. Method::Using the human FLS as the research object, the nontoxic concentration of FLS.FLS was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay for the follow-up experiment. The transwell migration, adhesion and transwell invasion test were used to detect the migration and adhesion of the different concentration of Ermiaosan on FLS, respectively. The expression of interleukin (IL)-1 beta of FLS supernatant was detected by enzyme linked immunosorbent assay (ELISA). Protein in FLS was extracted and protein expression levels of phosphorylated Janus kinase 1 (p-JAK1), p-signal transducer and activator of transcription (STAT1) and p-STAT6 were detected by Western blot. Result::Compared with control group, tumor necrosis factor(TNF)-α (20 μg·L-1) increased the proliferation, migration, adhesion, invasion and the secretion of IL-1β of FLS (P<0.01). Ermiaosan(0.2, 0.4, 0.8 mg·L-1) had no significant effect on the proliferation of FLS induced by TNF-α for 24 h. Within 24 h, the migration, adhesion, invasion, invasion, and secretion of IL-1β of FLS cells induced by TNF-α were also decreased significantly(P<0.05, P<0.01). Compared with the blank group, TNF-α could induce abnormal elevation of p-JAK1, p-STAT1 and p-STAT6 in FLS (P<0.01), while Ermiaosan of 0.2, 0.4, 0.8 g·L-1 could significantly reduce the expression levels of p-JAK1, p-STAT1 and p-STAT6 (P<0.05, P<0.01). Conclusion::Ermiaosan can inhibit the migration, adhesion and invasion of FLS, and its mechanism may be related to the inhibition of the secretion of IL-1β, the mechanism may be related to JAK/STAT pathway.
ABSTRACT
OBJECTIVE:To study the effect of Xuchangqing-ermiaosan-santeng formula on the contents of tumor necrosis fac-tor α(TNF-α)and ossification-related factor DKK-1 in serum of model mice with arthritis,and reveal its mechanism in the treat-ment of arthritis. METHODS:60 BALB/c mice were randomly divided into normal group,model group,sulfasalazine group(posi-tive control,9 mg/kg) and Xuchangqing-ermiaosan-santeng formula low-dose,medium-dose,high-dose groups (calculated by crude drug as 11.25,22.5,45 g/kg),10 in each group. Except for normal group,other 50 mice were intraperitoneallly injected complete Freund's adjuvant + proteoglycans to induce model with arthritis. After modeling,mice in each administration group were intragastrically administrated relevant medicines,mice in normal group and model group were intragastrically administrated equal volume of normal saline,once a day,for 20 d. After administration,enzyme-linked immunosorbent assay was used to detect the contents of TNF-αand DKK-1 in serum of mice in each group,and ultrastructural changes of sacroiliac joint synovial cells were ob-served by transmission electron microscopy. RESULTS:Compared with normal group,TNF-α content in serum in model group was obviously increased,DKK-1 content was obviously decreased (P<0.05);sacroiliac joint synovial cells showed hyperplasia, organellar deformation,mitochondrial swelling and other pathologic damage. Compared with model group,TNF-α contents in se-rum in each administration group were obviously decreased(P<0.05 or P<0.01);except for sulfasalazine group,the DKK-1 con-tent of mice in other administration groups were obviously increased (P<0.05). Pathologic damages of sacroiliac joint synovial cells in each administration group were reduced to varying degrees,and improvement degree in Xuchangqing-ermiaosan-santeng for-mula groups was higher than sulfasalazine group. CONCLUSIONS:Xuchangqing-ermiaosan-santeng formula may inhibit the sacroil-iac arthritis and pathological ossification of model mice with arthritis by decreasing TNF-α content and increasing DKK-1 content in serum.
ABSTRACT
BACKGROUND: Various health benefits have been attributed to Er-Miao-San (EMS), a traditional Chinese herbal formulation that contains equal amounts of cortex phellodendri (Phellodendron amurense Ruprecht) and rhizoma atractylodis (Atractylodes lancea D.C). However, its effect on the anti-inflammatory activity in human dermal fibroblasts (HDFs) and the mechanism underlying this effect are unknown. RESULTS: This study investigated the effects of EMS on TNF-α-induced MMP-1 expression in HDFs. Our data show that EMS inhibited TNF-α-induced MMP-1 expression in a concentration-dependent manner. Interestingly, EMS maintained IkB content without inhibiting the phosphorylation of MAPKs, which are well-established upstream kinases of NF-kB. Moreover, EMS reduced the level of nuclear p65 protein in HDFs. Luciferase assay revealed that EMS inhibits the transcriptional activity of NF-kBbystabilizing IkB. Our results show that EMS exerts its anti-inflammatory effect by inhibiting NF-kB-regulated genes such as IL-1ß and IL-8. Moreover, EMS effectively inhibited TNF-α-induced expression of MMP-1 via the NF-kBpathway. CONCLUSIONS: Taken together, our data suggest that EMS could potentially be used as an anti-inflammatory and anti-aging treatment.