Ethyl acetate fraction from Angelica sinensis inhibits IL-1ß-induced rheumatoid synovial fibroblast proliferation and COX-2, PGE2, and MMPs production

BACKGROUND: The root of Angelica sinensis (AS), also known as "Dang-gui," was a popular herbal medicine widely used in the treatment of gynecological diseases in China, Korea, and Japan for a long time. This study aimed to determine the effects of ethyl acetate fraction from Angelica sinensis (EAAS) on the interleukin-1ß (IL-1ß)-induced proliferation of rheumatoid arthritis synovial fibroblasts (RASFs), and production of matrix metalloproteinases (MMPs), cyclooxygenase (COX) 2, and prostaglandin E2 (PGE2), involved in articular bone and cartilage destruction, by RASFs. RESULTS: RASF proliferation was evaluated with cholecystokinin octapeptide (CCK-8) reagent in the presence of IL-1ß with/without EAAS. Expression of MMPs, tissue inhibitor of metalloproteinases-1 (TIMP-1), COXs, PGE2, and intracellular mitogen-activated protein kinase (MAPK) signaling molecules, including p-ERK, p-p38, p-JNK, and NF-κB, were examined using immunoblotting or semi-quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. EAAS inhibited IL-1ß-induced RASF proliferation; MMP-1, MMP-3, and COX-2 mRNA and protein expressions; and PGE2 production. EAAS also inhibits the phosphorylation of ERK-1/2, p38, and JNK, and activation of NF-κB by IL-1ß. CONCLUSION: EAAS might be a new therapeutic modality for rheumatoid arthritis management.

Immunohistochemical Characterization of Human Synovial Bursa Cells by Light and Transmission Electron Microscopy: Where do These Cells Come From? / Caracterización Inmunohistoquímica de las Células de la Bolsa Sinovial Humana Através de Microscopías de Luz y Transmisión: ¿De dónde Provienen esas Células?

In the present study human synovial bursa specimens were examined by light and transmission electron microscopy. For light microscopical investigation the bursa tissue was stained with azan, haematoxylin-eosin and monoclonal antibodies (CD14, CD33, CD36, CD68, laminin). For electron microscopical investigation the bursa specimens were fixated with Karnovsky's solution and 1,5% osmium tetroxide (Os0(4)) in water distilled and contrasted with 5% uranylacetate and embedded in Epon®. For the first time the antigenic phenotype was characterized and conclusions were drawn about the origin of the synovial bursa cells. Histologically the bursa was divided in two distinct layers; the intima, which is formed by a lining layer and a lamina propria, and a subintimal layer. The intima consisted of macrophage like (type I) and fibroblast like cells (type II). According to the immunohistochemical staining and the electron microscopy the type I cell seemed to be a bone marrow derived monocyte and the more frequently seen type II cell was derived from subintimal fibroblasts. The intimal bursa cell frequently interdigitated and usually communicated by their filopodia (indirect cell-cell-communication). Neither tight or gap junctions nor desmosomes could be documented. Although there was no evidence for the existence of a basal lamina, a concentration of extracellular matrix components beyond the bursa cells was observed. In our study there was no accumulation of laminin around the bursal cells, but striking was a vascular bundle of the intima subintima border zone, which was positive for laminin and CD68 and separated the intima from the subintima. In our opinion this histological structure plays an important role in the regeneration of the lining cells and acts like a barrier between bursa and blood.

En el presente estudio se examinaron bolsas sinoviales humanas a través de microscopía de luz y electrónica de transmisión. Para la microscopía de luz, el tejido de las bolsas se tiñó con Azan, H-E y anticuerpos monoclonales (CD14, CD33, CD36, CD68, laminina). Para la microscopía electrónica las bolsas fueron fijadas con solución de Karnovsky y tetróxido de osmio al 1,5% (Os04) en agua destilada y contrastada con acetato de uranilo al 5% y embebido en Epon®. En primera instada, el fenotipo antigénico fue caracterizado, concluyéndose acerca del origen de las células que componen la bolsa sinovial. Histológicamente la bolsa fue dividida en dos capas distintas - la íntima - la cual es formada por una capa lineal y una lámina propia, y, una subintima. La íntima consistió en células parecidas a macrófagos (Tipo I) y células semejantes a fibroblastos (Tipo II). De acuerdo a la tinción inmunohistoquímica y a la microscopía electrónica, las células tipo I parecen provenir de la médula ósea derivada de monocitos y el más frecuente tipo celular II fue derivadado de los fibroblastos de la subintima. Frecuentemente las células de la íntima de la bolsa se interdigitaban y usualmente se comunicaban a través de sus prolongaciones (comunicación célula indirecta-célula). No se observaron ni uniones abiertas, ni cerradas, ni desmosomas. Aunque no hubo evidencia de la existencia de una lámina basal, se observó una concentración de componentes de matriz extracelular más allá de las células de la bolsa. No hubo acumulación de laminina alrededor de estas células, pero destacada era una banda vascular de la zona límite entre íntima y subintima, la cual fue positiva para laminina y CD68 la cual separaba la íntima de la subintima. En nuestra opinión esta estructura histológica juega un importante rol en la regeneración de las células lineales y actúa como una barrera entre la bolsa y la sangre.