目的研究阿托伐他汀在体外对人CD4+T淋巴细胞张力蛋白同源第10染色体丢失的磷酸酶基因(胛EN)表达的影响。方法取25例健康志愿者的新鲜外周血,免疫磁珠分选出CD4+T淋巴细胞,随机分为空白组、植物血凝素(PHA)刺激组、PHA+1μmol/L阿托伐他汀组、PHA+5μmol/L阿托伐他汀组,PHA+10μmol/L阿托伐他汀组,体外培养48h后收集各组细胞及培养基上清液,荧光定量PCR检测PTENmRNA表达水平,Westernblot检测PTEN蛋白表达,ELISA检测培养基上清液肿瘤坏死因子d(TNF—a)、白细胞介素6(IL-6)及白细胞介素10(IL-10)浓度。结果与空白组比较,PHA刺激后,CD4+T淋巴细胞PTENmRNA、蛋白的表达及上清液TNF—d、IL-6浓度均升高(P〈0.05),而IL-10浓度升高无统计学差异(P〉0.05)。与PHA刺激组比较,PHA+5μmol/L阿托伐他汀组、PHA+10μmol/L阿托伐他汀组CD4+T淋巴细胞PTENmRNA、蛋白的表达和上清液IL-10浓度增加(P〈0.05),而PHA4-1μmol/L阿托伐他汀组具有增高趋势(P〉0.05),并随着阿托伐他汀药物浓度的增加而增加;各组上清液TNF.a、IL-6浓度降低,PHA+5μmol/L阿托伐他汀组、PHA-4-10μmol-/L阿托伐他汀组具有统计学差异(P〈0.05)。直线相关性分析显示,TNF-仪、IL-6的分泌水平与PTEN的表达量呈明显的负相关关系(r=-0.837和r=-0.816,P〈0.01),IL-10的分泌水平与PTEN的表达量呈明显的正相关关系(r=0.753,P〈0.05)。结论阿托伐他汀能够通过调控人cD4+T淋巴细胞PTEN表达发挥抗炎作用。
Aim To investigate the influence of atorvastatin on the expression of phosphatase and tensin homolog deleted on chromosome (PTEN) involved in human CIM + T lymphocytes in vitro. Methods Human CD4 + T cells obtained from healthy individuals were activated with phytohemagglutinin (PHA) and treated with atorvastatin. The levels of the inflammatory cytokines TNF-a, IL-6 and IL-10 were tested by enzyme-linked immunosorbent assay (ELISA). Re- al-time fluorescent quantitative polymerase chain reaction (RFQ-PCR) was used to measure the expression of PTEN mRNA in CIM + T cells, which were assessed at the protein level by Western blot as well. Results Compared to the blank group, the stimulation of PHA obviously increased the mRNA and protein expression of PTEN and those serum TNF-a and IL-6 secretion (P 〈 0.05 ), while the level of IL-10 was different between two groups ( P 〉 0.05 ). Compared with those of PHA stimulation group, different concentration atorvastatin significantly increased the protein level of PTEN expression and serum IL-10 secretion (P 〈 0. 05), while serum TNF-a and IL-6 production were reduced (P 〈 0.05 ). Serum TNF- a and IL-6 secretion were negatively related to the expression of PTEN through the linear related analysis ( r = - 0. 837 andr= -0.816, P〈0.01) , and IL-10 positive relation with the expression of PTEN (r =0.753, P〈0.05). Conclu- sion The anti-inflammatory effects of atorvastatin are mediated by regulating the expression of PTEN in CD4 + T cells.