Department of Ophthalmology, Beijing Chaoyang Hospital, Capital Medical University, No. 8 Baijiazhuang Road Chaoyang District, Beijing 100020, China.
Biomed Res Int. 2013;2013:431232. doi: 10.1155/2013/431232. Epub 2013 Sep 16.
To evaluate the effect of a traditional Chinese medicine, Rheum Polysaccharide (RP), on the in vitro production of tumor necrosis factor alpha (TNF- α ) and interleukin-10 (IL-10) by lipopolysaccharide- (LPS-)stimulated human monocytes from HLA-B27 associated acute anterior uveitis patients of convalescence stage.
PBMC samples were isolated from 10 HLA-B27 associated acute anterior uveitis, incubated, respectively, and divided into 4 groups as follows: (1) controls, PBS was added in final concentration of 1 mg·L⁻¹, (2) stimulated by LPS, LPS was added in final concentration of 1 mg·L⁻¹, (3) stimulated by LPS + HTA125, 30 minutes before the adding of LPS in final concentration of 1 mg·L⁻¹, the final concentration of 5 mg·L⁻¹ of the HTA125 was added, and (4) stimulated by LPS + RP, 30 minutes before the adding of LPS in final concentration 1 mg·L⁻¹, the final concentration 100 mg·L⁻¹ of the RP was added. Supernatants were used to quantify the amounts of TNF- α and IL-10 released in time course using enzyme-linked immunosorbent assay (ELISA).
After stimulated by lps, the concentrations of TNF- α and IL-10 in culture supernatants of patients are significantly higher than control group at all time points (P < 0.01). Blockage of TLR-4 by HTA125 can decrease the production of TNF- α and IL-10 compared with lps group (P < 0.01, except at 4 h group of IL-10). Concentration of TNF- α and IL-10 also decreases in the LPS + RP group (P < 0.01) but not so significantly as in the LPS + HTA125 group.
As anti-TLR4 monoclonal antibodies, rheum Polysaccharide can also inhibit the secretion of cytokines produced by monocytes from HLA-B27 positive AAU patients of convalescence stage.
评估中药白芍总苷(RP)对恢复期 HLA-B27 相关性急性前葡萄膜炎患者脂多糖(LPS)刺激的人单核细胞肿瘤坏死因子-α(TNF-α)和白细胞介素-10(IL-10)体外产生的影响。
从 10 例 HLA-B27 相关性急性前葡萄膜炎患者中分离 PBMC 样本,孵育并分为 4 组:(1)对照组,终浓度为 1mg·L⁻¹加入 PBS;(2)LPS 刺激组,终浓度为 1mg·L⁻¹加入 LPS;(3)LPS+HTA125 刺激组,在终浓度为 1mg·L⁻¹加入 LPS 前 30 分钟,终浓度为 5mg·L⁻¹加入 HTA125;(4)LPS+RP 刺激组,在终浓度为 1mg·L⁻¹加入 LPS 前 30 分钟,终浓度为 100mg·L⁻¹加入 RP。采用酶联免疫吸附试验(ELISA)测定时间过程中上清液中 TNF-α和 IL-10 的释放量。
LPS 刺激后,患者培养上清液中 TNF-α和 IL-10 的浓度在所有时间点均明显高于对照组(P<0.01)。与 LPS 组相比,TLR-4 阻断剂 HTA125 可降低 TNF-α和 IL-10 的产生(P<0.01,除 IL-10 的 4h 组外)。LPS+RP 组 TNF-α和 IL-10 浓度也降低(P<0.01),但不如 LPS+HTA125 组显著。
作为抗 TLR4 单克隆抗体,白芍总苷还可以抑制恢复期 HLA-B27 阳性 AAU 患者单核细胞产生细胞因子。