rhIL-11对中子辐射IEC-6细胞IL-11Rα和gp130表达的影响

Effect of rhIL-11 on expressions of interleukin-11 receptor α-chain and gp130 in IEC-6 cells after neutron irradiation

目的探讨rhIL-11对中子照射后IEC-6细胞IL-11Rα和gp130表达的影响。方法培养的IEC-6细胞经4·0Gy中子照射并于照前12h与照后即刻给予100ng/ml的rhIL-11,采用流式细胞术、免疫组化、Westernblot及图像分析等技术检测IEC-6细胞的凋亡和坏死率、IL-11Rα和gp130的表达。结果中子照射后6h,IEC-6细胞凋亡率增加(P<0·01),应用rhIL-11可以降低细胞凋亡率(P<0·05)。IL-11Rα于正常IEC-6细胞膜和浆呈强阳性,中子照射后6h,IL-11Rα表达明显减少(P<0·01),24h恢复至正常水平(P<0·01),应用rhIL-11后IL-11Rα表达高于单纯照射组(P<0·05)。gp130于正常IEC-6细胞浆呈强阳性,中子照射后48h内呈进行性减少(P<0·05),应用rhIL-11后gp130表达于照射后24h恢复正常,48h降低但高于单纯照射组(P<0·05)。结论IL-11对IEC-6细胞中子辐射损伤起保护作用,其机制可能是通过上调其特异性结合受体亚基IL-11Rα和信号转导亚基gp130发挥作用。

Objective To observe the effect of recombinant human interleukin-11 （rhlL-11） on expressions of interleukin-11 receptor α-chain （IL-11Rα） and glycoprotein 130 （gp130）, an additional signal transducer, in IEC-6 ceils after neutron irradiation. Methods IEC-6 cells were exposed to 4.0 Gy neutron and administered with 100 ng/ml rhIL-11 12 h before or immediately after irradiation. The apoptosis and necrosis rates and expressions of IL-11Rα and gp130 were detected by flow cytometry, immunohistochemistry, Western blot, and image analysis. Results The apoptosis rate of IEC-6 cells was increased by irradiation at 6 h （ P 〈 0.01 ）, IL-11 stimulation resulted in a decreased apoptosis rate in irradiated IEC-6 cells （P 〈 0.05）. In normal control IEC-6 ceils, IL-11 Rα was strongly expressed in the cellular membrane and cytoplasm. The level of IL-11 Rα expression significantly decreased at 6 h after irradiation （ P 〈 0.01 ） and returned to the normal level at 24 h after the irradiation. In IEC-6 cells treated with both radiation and rhIL-11, the level of IL-1 IRa expression was higher than that of the irradiated cells （ P 〈 0.05）, while gp130 protein was strongly expressed in the cytoplasm of IEC-6 ceils. After irradiation, we found a progressive decrease in the expression of gp130 protein （ P 〈 0.05） in 48 h, while in rhIL-11-stimulated cells, it returned to the normal level at 24 h after irradiation and decreased at 48 h, but still higher than that of the irradiated-only cells （ P 〈 0.05 ）. Conclusions rhlL- 11 can protect IEC-6 cells from neutron irradiation damage. The protection effect of rhIL-11 might be connected with its ability of up-regulating expressions of specific ligand-binding subunit IL-1 IR Neutron; IEC-6; Interleukin-11; IL-11Rα; gpl30 and signal-transducing subunit gp130.