[乌梢蛇II型胶原蛋白对佐剂性关节炎大鼠滑膜细胞细胞因子产生的影响]

[The effect of the type II collagen protein from Zaocys on cytokines production by synoviocytes in rats with adjuvant arthritis].

作者信息

Pang Jie, Li Juan, Wu Xiang-Hui, Wu He-Yong

机构信息

Department of Rheumatic, Southern Medical University, Guangzhou 510515, China.

出版信息

Zhong Yao Cai. 2009 Apr;32(4):556-60.

PMID:19645241

Abstract

OBJECTIVE

To investigate the effect of the type II collagen (C II) protein from Zaocys on cytokines production by synoviocytes in rats with adjuvant arthritis (AA).

METHODS

Type II collagen protein was abstracted and purificated from Zaocys. Adjuvant arthritis (AA) was induced by a single intradermal injection of 0.1 mL of complete Freund's adjuvant into the left hind paw. Synoviocytes' supernatants were harvested and synoviocytes-Peyer's Patches (PP) cells coculture system were developed. Tumor necrosis factor-alpha (TNF-alpha) activity was measured by L929 cytotoxicity bioassay and Interleukin (IL)-1beta activity was measured by MTT dye reduction. The synoviocytes' supernatants cytokines' levels were detected by ELISA.

RESULTS

Each concentration of C II from Zaocys had no effect on IL-1beta and TNF-alpha production by synoviocytes in vitro. Middle concentration of C II suppressed the activity of IL-1beta and TNF-alpha production by synoviocytes-PP cells coculture system (P < 0.05). Treating with low and high dose of C II suppressed the activity of TNF-alpha and IL-1beta producing by synoviocyte (P < 0.05), significantly suppressed in the group of AA rats treated with middle dose of C II (P < 0.01). Treating with middle and high dose of C II decreased the level of synoviocytes' supernatants TNF-alpha (P < 0.05), the level of synoviocytes' supernatants IL-1beta decreased in all treating groups (P < 0.05). Treating with middle dose of C II increased the level of serum TGF-beta (P < 0.05). Middle concentration of C II suppressed the activity of IL-1 and TNF production by synoviocytes-PP cells coculture system (P < 0.05).

CONCLUSIONS

C II from Zaocys has no direct effect on the activity of IL-1beta and TNF production by synoviocytes in vitro. Oral administration of type II collagen protein from Zaocys can effectively suppressed the activity and level of the cytokines production by synoviocytes in rats with adjuvant arthritis (AA).

摘要

目的

研究乌梢蛇中Ⅱ型胶原蛋白（CⅡ）对佐剂性关节炎（AA）大鼠滑膜细胞产生细胞因子的影响。

方法

从乌梢蛇中提取并纯化Ⅱ型胶原蛋白。通过在左后爪皮内单次注射0.1 mL完全弗氏佐剂诱导佐剂性关节炎（AA）。收集滑膜细胞的上清液并建立滑膜细胞-派伊尔结（PP）细胞共培养体系。采用L929细胞毒性生物测定法测定肿瘤坏死因子-α（TNF-α）活性，采用MTT比色法测定白细胞介素（IL）-1β活性。通过酶联免疫吸附测定法检测滑膜细胞上清液中细胞因子的水平。

结果

各浓度的乌梢蛇CⅡ对体外培养的滑膜细胞产生IL-1β和TNF-α均无影响。中浓度的CⅡ抑制滑膜细胞-PP细胞共培养体系中IL-1β和TNF-α的产生活性（P<0.05）。低剂量和高剂量的CⅡ处理可抑制滑膜细胞产生TNF-α和IL-1β的活性（P<0.05），在中剂量CⅡ处理的AA大鼠组中抑制作用显著（P<0.01）。中剂量和高剂量的CⅡ处理可降低滑膜细胞上清液中TNF-α的水平（P<0.05），所有处理组中滑膜细胞上清液中IL-1β的水平均降低（P<0.05）。中剂量的CⅡ处理可提高血清转化生长因子-β（TGF-β）的水平（P<0.05）。中浓度的CⅡ抑制滑膜细胞-PP细胞共培养体系中IL-1和TNF的产生活性（P<0.05）。