L—精氨酸和尼莫地平对脂多糖诱导的内皮细胞表达单核细胞趋化蛋 …

目的 观察Ｌ－精氨酸和尼莫地平对脂多糖（ＬＰＳ）诱导的内皮细胞（ＥＣ）的单核细胞趋化蛋白－１（ＭＣＰ－１）ｍＲＮＡ和蛋白表达的影响。方法 在ＥＣ培养基中分别为ＬＰＳ，ＬＰＳ＋Ｌ－精氨酸和ＬＰＳ＋尼莫地平，培养８小时，用异硫氢酸胍法提了细胞的总ＲＮＡ，用γ＾２－３２Ｐ末端标记的ＭＣＰ－１寡核苷酸探针培养基（ＣＭ）中的ＭＣＰ－１蛋白含量，用免疫细胞化学方法检测ＥＣ的ＭＣＰ－１的蛋白的表达。结果 斑点杂

The effects of L-arginine and nimodipine on the expression of monocyte chemoattractant protein-1 in endothelial cells induced by lipopolysaccharide]

OBJECTIVE: To examine the effects of L-arginine and nimodipine on the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein induced by lipopolysaccharide (LPS) in endothelial cells (ECs). METHODS: After 8-hour exposure to LPS, LPS + L-arginine and LPS + nimodipine respectively, the total RNA in ECs was extracted by the guanidinium isothiocyanate method. The MCP-1 mRNA expression in ECs was examined by Dot blot analysis using a gamma-32P-end-labeled 35 mer oligonucleotide probe of MCP-1. MCP-1 protein in the EC-conditioned media (EC-CM) was determined by sandwich ELISA for each group. The expression of MCP-1 protein in ECs was examined immunocytochemically. RESULTS: Dot blotting showed that after exposure to LPS, the integral absorbance (A) value of the dots of MCP-1 mRNA on the nitrocellulose membrane were 3.14, which was 2.5-fold as much as that in the control group (A = 1.24). The A values of the LPS + L-arginine group and the LPS + nimodipine group were 1.43 and 1.65, respectively. ELISA showed that the MCP-1 protein content in the EC-CM of the LPS group was 6.29 +/- 0.53 ng/ml, which was markedly higher than that of the control group (2.32 +/- 0.16) ng/ml (P < 0.01). The MCP-1 protein content in the EC-CM of the LPS + L-arginine group and the LPS + nimodipine group was significantly lower than that of the LPS group (P < 0.01). Furthermore, the ECs in LPS group showed a strong immunoreactivity for the polyclonal MCP-1 antibody, whereas the ECs in control, L-arginine and nimodipine groups revealed a weak postive immunostaining. CONCLUSION: L-arginine and nimodipine can markedly inhibit the expression of MCP-1 in ECs induced by LPS.