藍色忍冬屬植物萃取物作用於 LPS(脂肪多醣類 )所導致的發炎反應

Effects of blue honeysuckle (Lonicera

caerulea L.) extract on lipopolysaccharide-

induced inflammation in vitro and in vivo

作者： Jin, X.H., Ohgami, K., Shiratori, K., Suzuki, Y.,

Koyama, Y.,
Yoshida, K., Ilieva, I., Tanaka, T., Onoe, K.and Ohno, S.
出處： Eye Research 82 (2006) 860–867

演講者：蔡一誠

日期：

書面摘要：

本研究利用葡萄膜炎的作用 (EIU,end oto xin-in duced uve itis ) 的老鼠

來研究忍冬屬植物萃取物 (BHE)的作用，我們發現 BHE影響了細胞的滲
透作用、蛋白質外滲、 TNF-α、PGE2與NO濃度。為了證實其抗發炎反應，
我們觀察經過 BHE處理的老鼠虹膜睫體中的 NF-κB。另外，在經過 BHE處
理的 RAW264.7細胞 (老鼠巨噬細胞細胞線 )觀察了 NO、腫瘤壞死因素
(TNF-α)、和前列腺素 (PGE2)、COX-2和iNOS的濃度。
在老鼠體內注射脂肪多醣類 (LPS, lipopolysaccharide)導致了
EIU。注射 LPS後，再將 1、10、100 mg的BHE進行靜脈注射。 在老鼠體液
組注射 LPS經過 24H之後，觀察滲入細胞的數量、蛋白質含量、氧化物
(NO)、腫瘤壞死因素 (TNF-α)、和前列腺素 (PGE2)。結果顯示，在體液組
中發現 BHE大量減少了發炎細胞滲透活動、 蛋白質含量、 NO濃度、TNF-α
與PGE2，且明顯地改善了眼睛組織狀態。
BHE有效地壓制 LPS對RAW264.7細胞所誘增的 NO、PGE2和TNF-
α，及大大降低了 iNOS和COX-2的濃度。藉著 BHE抑制 NF-κB和發炎因子 ，
使EIU在眼睛誘導的發炎程度而降低。 觀察 BHE影響 NF-κB的作用，以進
一步評估抗發炎效果。

The aim of the present study was to investigate the

effects of blue honeysuckle extract (BHE), which contains
high level of phenolic compounds, on endotoxin-induced
uveitis (EIU). Male Lewis rats were randomly divided into 5
groups with 14 rats in each (eight rats for collection of
aqueous humor, six rats for histologic examination). EIU was
induced by a footpad injection of lipopolysaccharide (LPS).
1, 10, or 100 mg of BHE was injected intravenously
immediately after LPS injection. The aqueous humor was
collected at 24 h after LPS injection, the number of
infiltrating cells, protein concentration, nitric oxide (NO),
tumor necrosis factor (TNF)-a, and prostaglandin (PG)-E2
levels in the aqueous humor were determined.
Some eyes were enucleated for histologic examination
and immunohistochemical analysis. Immunohistochemical
staining with a monoclonal antibody against activated
nuclear factor (NF)-kB was performed to evaluate the effect
of BHE on NF-kB activation. To further clarify the anti-
inflammatory effect, RAW264.7 cells (a mouse macrophage
cell line) were stimulated with LPS in the presence or
absence of BHE and its major phenolics, cyanidin 3-glucoside
(C3G), cyanidin 3-rutinoside (C3R), chlorogenic acid (CA).
Expression of inducible NO synthase (iNOS) and
cyclooxygenase-2 (COX-2) were analyzed by Western blot
method.
BHE treatment significantly reduced the inflammatory
cell infiltration, the protein concentration, the levels of NO,
TNF-a and PGE2 in the aqueous humor and improved
histologic status of the ocular tissue. The number of
activated NF-kB-positive cells was lower in the iris-ciliary
body treated with BHE at 3 h after LPS injection.
BHE significantly suppressed the production of NO, PGE2
and TNF-a in the culture medium as well as the expression
of iNOS and COX-2 by LPS-stimulated RAW264.7 cells in a
dose-dependent fashion. C3G, C3R and CA showed no or
weak inhibitory effects on the level of inflammatory
mediators and the expression of iNOS and COX-2.
These results suggest that BHE attenuates the degree of
inflammation in eyes with EIU by inhibiting the NF-kB
dependent signaling pathway and the subsequent
production of proinflammatory mediators.