脂多糖LPS对糖尿病大鼠肺泡巨噬细胞RBD-2表达的影响

目的以肺泡巨噬细胞为研究对象,观察脂多糖（lipopolysaccharide,LPS）刺激下RBD-2（大鼠β防御素-2,Rat-βdefensins-2）在正常大鼠及糖尿病大鼠肺泡巨噬细胞表达的变化。方法以健康雄性SD大鼠制备糖尿病模型。32只大鼠随机分为四组：正常对照组（A组）、糖尿病组（B组）、LPS组（C组）和糖尿病＋LPS组（D组）,每组均为8只。分离培养大鼠肺泡巨噬细胞,通过RT-PCR以及Western blotting分别检测RBD-2的RNA及蛋白质表达水平,同时通过Real Time PCR检测大鼠肺泡巨噬细胞的TLR-2以及TLR-4的mRNA的表达。结果大鼠糖尿病模型构建成功。RT-PCR以及Western blotting结果显示,与正常组相比,糖尿病组、正常组＋LPS组、糖尿病＋脂多糖组的RBD-2 mRNA以及蛋白质表达水平依次增加,差异显著（P〈0.05）。Real Time PCR结果显示,与正常组相比,糖尿病组、正常组＋LPS组、糖尿病＋脂多糖组的TLR-4 mRNA的表达水平依次增加,差异显著（P〈0.05）,而TLR-2差异则不明显。结论 LPS刺激后糖尿病大鼠肺泡巨噬细胞RBD-2表达增加较糖尿病组更为显著,说明RBD-2变化对于增强糖尿病机体的非特异性免疫能力具有明显的帮助,同时糖尿病大鼠肺泡巨噬细胞RBD-2表达较正常组增高,说明处于糖尿病时期的大鼠机体处于炎症状态,并且这一通路的表达受体主要是TLR-4受体。

Expression of RBD-2 in alveolar macrophages of diabetic rats regulated by LPS

Objective To observe the expression of rat β-defensin 2（RBD-2） in alveolar macrophages of diabetic rats after lipopolysaccharide（LPS） stimulation.Methods Healthy SD rats were used to construct the diabetic models.Then they were randomly divided into four groups.Group A： the control group;Group B： the diabetic group;Group C： the LPS stimulated group;Group D： the diabetic group with LPS infection.Rat alveolar macrophages were captured and cultured.The RT-PCR and Western blotting method were utilized to detect the mRNA and protein level of RBD-2.The Real Time PCR method were used to determine the mRNA level of TLR-2 and TLR-4.Results The results of RT-PCR and Western blotting indicated that the expression level of RBD-2 mRNA and protein in group B,group C and group D followed by increased significantly（P〈0.05）.Subsequently,the results of Real Time PCR indicated that the expression level of TLR-4 mRNA in group B,group C and group D followed by increased significantly（P〈0.05） but that of the TLR-2 was different.Conclusion The expression of RBD-2 is obvious after LPS stimulation,which suggests that RBD-2 help to enhance the nonspecific immunity in diabetic rat.Subsequently,the expression of RBD-2 of diabetic rats also indicates that diabetic bodies are in the pro-inflammatory state and the main receptor was TLR-4.