博来霉素刺激外周血单核细胞因子引起的成纤维细胞增殖。

Fibroblast proliferation by bleomycin stimulated peripheral blood mononuclear cell factors.

作者信息

Yamamoto T, Katayama I, Nishioka K

机构信息

Department of Dermatology, Tokyo Medical and Dental University, School of Medicine, Japan.

出版信息

J Rheumatol. 1999 Mar;26(3):609-15.

PMID:10090171

Abstract

OBJECTIVE

To evaluate the role of bleomycin for cutaneous fibrosis, we investigated the effects of bleomycin stimulated peripheral blood mononuclear cell (PBMC) factors on in vitro proliferation of mouse 3T3 fibroblasts.

METHODS

PBMC were incubated with 10(-4) to 10 microg/ml of bleomycin, and the supernatants (conditioned media) were collected after 48 h. 3H-TdR incorporation was assayed to examine the effects of conditioned medium on 3T3 fibroblast proliferation, which was also compared between 8 patients with systemic sclerosis (SSc) and 8 control subjects.

RESULTS

Conditioned medium stimulated 3T3 fibroblasts to proliferate in a dose dependent manner, while culture supernatant without stimulation of bleomycin showed no growth stimulatory activity. Conditioned medium derived from PBMC of patients with SSc exhibited a significantly higher growth activity than that from controls (p < 0.05). Among PBMC subpopulations, macrophages showed the most increased growth effect on 3T3 fibroblasts, although T lymphocyte populations also showed growth activity. The growth activity was inhibited up to 35% by anti-platelet derived growth factor (PDGF) antibody. In addition, growth activity was also partially blocked by antibodies against basic fibroblast growth factor (bFGF) (20%), interleukin 1beta (IL-1beta) (15%), tumor necrosis factor-alpha(TNF-alpha) (7.9%), and transforming growth factor-beta (TGF-beta)(9.5%). The cellular release of IL-1beta, TNF-alpha, and IL-6 was elevated in response to bleomycin exposure in both controls and patients with SSc. Reverse transcriptase-polymerase chain reaction revealed that conditioned medium derived from both patients with SSc and controls induced mRNA expression of several fibrogenic cytokines such as IL-6, TNF-alpha, TGF-beta, and PDGF. IL-1alpha was induced only by conditioned medium derived from patients with SSc.

CONCLUSION

These results suggest that bleomycin induced cutaneous fibrosis as well as pulmonary fibrosis may be induced by inflammatory cells through several fibrogenic cytokines.

摘要

目的

为评估博来霉素在皮肤纤维化中的作用，我们研究了博来霉素刺激的外周血单个核细胞（PBMC）因子对小鼠3T3成纤维细胞体外增殖的影响。

方法

将PBMC与10⁻⁴至10微克/毫升的博来霉素孵育，48小时后收集上清液（条件培养基）。采用³H-TdR掺入法检测条件培养基对3T3成纤维细胞增殖的影响，并在8例系统性硬化症（SSc）患者和8例对照受试者之间进行比较。

结果

条件培养基以剂量依赖性方式刺激3T3成纤维细胞增殖，而未用博来霉素刺激的培养上清液无生长刺激活性。SSc患者PBMC来源的条件培养基显示出比对照组显著更高的生长活性（p < 0.05）。在PBMC亚群中，巨噬细胞对3T3成纤维细胞的生长促进作用增加最为明显，尽管T淋巴细胞群体也显示出生长活性。抗血小板衍生生长因子（PDGF）抗体可将生长活性抑制高达35%。此外，抗碱性成纤维细胞生长因子（bFGF）（20%）、白细胞介素1β（IL-1β）（15%）、肿瘤坏死因子-α（TNF-α）（7.9%）和转化生长因子-β（TGF-β）（9.5%）的抗体也部分阻断了生长活性。在对照组和SSc患者中，博来霉素暴露后IL-1β、TNF-α和IL-6的细胞释放均升高。逆转录聚合酶链反应显示，SSc患者和对照组来源的条件培养基均诱导了几种促纤维化细胞因子如IL-6、TNF-α、TGF-β和PDGF的mRNA表达。IL-1α仅由SSc患者来源的条件培养基诱导。