雷公藤甲素对IFN-γ刺激后人眼球后成纤维细胞的影响

Effects of triptolide on cultured human retroocular fibroblasts induced by interferon-γ

目的观察中药雷公藤提取物雷公藤甲素(triptolide,TL)对干扰素-γ(interferon-γ,IFN-γ)刺激后人眼球后成纤维细胞(retroocular fibroblasts,RF)表达CD40、合成透明质酸(hyaluronic acid,HA)和Ⅰ型胶原蛋白的变化以及对细胞增生的影响。方法体外培养的正常人RF分别与IFN-γ(100U.L-1)、TL孵育后,MTT比色法检测TL对RF增生的影响,放射免疫法检测HA和Ⅰ型胶原蛋白的合成情况,流式细胞仪检测CD40的表达情况。响,放射免疫法检测HA和Ⅰ型胶原蛋白的合成情况,流式细胞仪检测CD40的表达情况。结果TL能明显抑制RF的增生,当剂量达1.00μg·L-1时抑制率为63.38%;当剂量达10.00μg·L-1时,抑制率达79.74%;当剂量达100.00μg·L-1时,RF增生完全被抑制。IFN-γ能刺激HA和Ⅰ型胶原蛋白的合成(P均<0.05)以及CD40的表达(P<0.01),HA和Ⅰ型胶原蛋白合成量及CD40阳性细胞百分率分别是对照组的1.64、1.72和13.27倍,而TL能明显抑制IFN-γ对RF的刺激作用,且随着剂量的增加,抑制作用更加明显。结论细胞因子IFN-γ能够增加HA和Ⅰ型胶原蛋白的分泌,刺激免疫调控分子CD40的过度表达,TL能够抑制RF的增生,抑制IFN-γ刺激后HA和Ⅰ型胶原蛋白的合成以及CD40的过度表达。

Objective To evaluate the effects of triptolide （ TL）, extracted from traditional Chinese drug Tripterygium Wiffordii, on cell proliferation, the synthesis of hyaluronic acid and collagen type I, and the expression of CD40 induced by interferon （IFN-γ） on the surface of cultured normal human retroocular fibroblasts（RF） in vitro. Methods After incubated with IFN-γ（ 100 U · L^-1 ） or TL, the proliferation of cultured normal RF in vitro was assessed using MTT colorimetry, the synthesis of hyalutonic acid and collagen type I were measured with radioimmunoassay, and the cell-surface expression of CD40 was examined by flow cytometry. Results The proliferation of RF was inhibited obviously by TL in a dose-dependent manner. The inhibition ratio was 53.38% with the concentration of 1.00μg·L^-1,79.74% with the concentration of 10.00 μg·L^-1 and was inhibited entirely with the concentration of 100.00 μg·L^-1. IFN-γ could stimulate the synthesis of hyaluronic acid and collagen type I （P 〈 0.05 ）, and enhance the expression of CD40（P 〈0.01 ） ,which was 1.54,1.72 and 13.27 times to control group, respectively. TL could antagonize the inflammatory effects of IFN-γ on RF significantly in a dose-dependent manner. Conclusion IFN-γ can increase the synthesis of HA and collagen type I and regulate the overexpression of CD40 TL, which can be inhibited by TL. TL inhibits the activation of cultured normal human RF in vitro induced by IFN-γ.