干扰素γ与甲强龙联合应用对人胚肺成纤维细胞增殖的影响★
李雪梅1，翟丽东2
相关链接：最近研究认为特发性间质性肺疾病是以纤维化为始发而非炎症诱导的，因此人们将关注的焦点由传统的抗炎治疗转向抗纤维化治疗。干扰素γ是一种抗纤维化的细胞因子，近年来在间质性肺疾病治疗方面的应用前景成为关注的热点。
应用要点：实验首次从细胞水平观察了干扰素γ与甲强龙联合应用对成纤维细胞增殖的影响，为进一步的动物实验及临床研究提供了理论基础。干扰素γ很可能成为治疗肺纤维化最有前景的药物，与甲强龙联合应用可能成为治疗肺纤维化最佳方案。�
术语解析：干扰素γ是由抗原特异性细胞和自然杀伤细胞产生，具有高度生物活性的一种细胞因子，是重要的免疫调节因子，具有对抗白细胞介素1、肿瘤坏死因子α，调节白细胞介素6合成等多种生物功能。具有生物活性的干扰素γ是非共价结合的同种二聚体，通过与特异性受体的相互作用，可在体内发挥广泛的生物学效应。
摘要
目的：目前间质性肺疾病的发病机制尚不完全清楚。实验拟进一步验证干扰素γ与甲强龙联合应用对人胚肺成纤维细胞增殖的影响，探究可否成为间质性肺疾病的有效治疗方法之一。
方法：实验于2005-07/2006-05在河北医科大学细胞研究室完成。①实验分组：选用第8代人胚肺成纤维细胞进行体外培养。参考两药人体常用药物剂量范围，选取细胞单独用药浓度将实验分为对照组、甲强龙组（15 mg/L）、干扰素γ（250，500，750，1 000，1 250 U/mL）组及甲强龙与γ干扰素联合应用组。②实验方法：不同浓度的干扰素γ与甲强龙单独或联合应用于人胚肺成纤维细胞 48 h。③实验评估：应用四甲基偶氮唑盐法观察人胚肺成纤维细胞的抑制率；应用流式细胞技术观察人胚肺成纤维细胞的细胞周期变化并计算增殖指数；应用免疫细胞化学方法检测增殖细胞核抗原的表达。
结果：①甲强龙和干扰素γ对人胚肺成纤维细胞均有抑制作用（P < 0.05），两药联合应用具有相加乃至协同作用(Q > 0.85)。②干扰素γ主要作用于G1和G2 M 期，甲强龙作用于S期，两药联合应用后细胞的增殖指数较两药单独应用降低（P < 0.05）。③甲强龙与不同浓度干扰素γ单独或联合应用均具有抑制人胚肺成纤维细胞增殖细胞核抗原表达的作用（P < 0.05），两药联合应用抑制增殖细胞核抗原表达作用增强（P < 0.05）。
结论：甲强龙及干扰素γ均可抑制人胚肺成纤维细胞增殖，两药联合应用具有协同作用。干扰素γ与甲强龙联合应用的抗纤维化效果可能优于两药单独应用。
关键词：人胚肺成纤维细胞；干扰素γ；甲强龙；增殖细胞核抗原；组织构建
李雪梅，翟丽东. 干扰素γ与甲强龙联合应用对人胚肺成纤维细胞增殖的影响[J].中国组织工程研究与临床康复，2008，12(15):2829-2833 [www.zglckf.com/zglckf/ejournal/upfiles/08-15/15k-2829(ps).pdf]
天津医科大学， 1代谢病医院足病科， 2解剖学教研室，天津市
300070
李雪梅★，女，1976年生，天津市人，汉族，2006年河北医科大学毕业，硕士，医师，主要从事间质性肺疾病的研究。
中图分类号:R392.114
文献标识码:A
文章编号:1673-8225
(2008)15-02829-05
收稿日期：2007-09-13 修回日期：2007-12-15
(07-50-9-5024/WL·A)
Influence of interferon-gamma in combination with methylprednisolone on the proliferation of human embryonic lung fibroblast
Abstract
AIM: The pathogenesis of interstitial lung disease is still uncertain. This study observed the effect of interferon-gamma (IFN-γ) in combination with methylprednisolone (M-pred) on the proliferation of human embryonic lung fibroblast (HELF) to identify whether it can be used as an effective treatment of interstitial lung disease in clinic.
METHODS: The experiment was performed at the Research Laboratory of Cytobiology, Hebei Medical University from July 2005 to May 2006. ①The eighth passage HELFs were cultured in vitro. According to the common dose range for human of IFN-γ and M-pred, cells were divided into control group, M-pred (15 mg/L) group, IFN-γ (250, 500, 750, 1 000, 1 250 U/mL) groups and combination group (IFN-γ and M-pred). ②HELFs were incubated with IFN-γ alone or combined with M-pred for 48 hours before they were harvested. ③MTT assay was used to measure the inhibition rate of M-pred combined with different concentrations of IFN-γ on HELF. Flow cytometry was used to observe cell cycle distribution and the proliferation index of HELF. The expression of proliferating cell nuclear antigen (PCNA) was detected by immunocytochemistry.
RESULTS: ①Both alone and combination usage of IFN-γ and M-pred could inhibit HELF proliferation (P < 0.05), and there was promotion, even synergistic effect in combination group (Q > 0.85). ② IFN-γ mainly acted on G1 and G2M periods, while M-pred on S period. The proliferation index decreased in combination group (P < 0.05). ③Both alone and combination usage of IFN-γ and M-pred could inhibit PCNA expression (P < 0.05), and the inhibitory effect was stronger in IFN-γ and M-pred combination group (P < 0.05).
CONCLUSION: Both IFN-γ and M-pred can inhibit the proliferation of HELF. There is synergism in the combination of IFN-γ and M-pred. The anti-fibrous degeneration effect of combination usage may be better than alone usage of IFN-γ or M-pred.
Li XM, Zhai LD. Influence of interferon-gamma in combination with methylprednisolone on the proliferation of human embryonic lung fibroblast.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(15):2829-2833(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-15/15k-2829(ps).pdf]
1Department of Diabetic Foot, Metabolic Disease Hospital, 2Department of Anatomy, Tianjin Medical University, Tianjin 300070, China
Li Xue-mei★, Master, Physician, Department of Diabetic Foot, Metabolic Disease Hospital, Tianjin Medical University, Tianjin 300070, China
Received:2007-09-31 Accepted: 2007-12-15
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