阿托伐他汀对草酸钙晶体诱导的细胞炎症反应的影响及其机制探讨

目的探讨阿托伐他汀(Atorvastatin,ATV)对草酸钙晶体诱导的人肾小管上皮细胞(HK-2细胞)炎症反应的影响及其作用机制。方法取HK-2细胞分为对照组(正常培养基培养)、ATV组(40μmol/LATV预处理3h后换成正常培养基)、草酸钙晶体刺激组(含4mmol/L草酸钙晶体的培养基)和ATV干预组(40μmol/LATV预处理3h后,换成含4mmol/L草酸钙晶体的培养基)。培养12h后收集细胞,采用免疫组化染色检查和蛋白质印迹法检测Nod样受体蛋白3(NLRP3)和Cleavedcaspase-1的表达水平;采用免疫荧光染色法和蛋白质印迹法检测NF-κB的表达水平;收集细胞培养上清液,采用酶联免疫吸附测定法(ELISA)检测白细胞介素(IL)-1β和IL-18的浓度。结果蛋白质印迹法检测结果显示,ATV组较对照组NLRP3(0.125±0.013与0.135±0.007)和Cleaved caspase-1(0.090±0.014与0.095±0.006)的相对表达量有所减少,但差异均无统计学意义(P>0.05);草酸钙晶体刺激组较对照组NLRP3(0.315±0.021与0.135±0.007,P<0.001)和Cleavedcaspase-1(0.235±0.008与0.095±0.006,P<0.001)的相对表达量明显增多;ATV干预组较草酸钙晶体刺激组NLRP3(0.245±0.007与0.315±0.021,P<0.05)和Cleavedcaspase-1(0.170±0.017与0.235±0.008,P<0.05)的相对表达量明显下降。免疫组化染色检查结果显示各组的NLRP3和Cleavedcaspase-1表达趋势与蛋白质印迹法检测结果一致。ELISA结果显示,ATV组较对照组细胞培养液中炎症因子IL-1β[(162.00±21.21)pg/ml与(183.50±7.78)pg/ml,P>0.05]和IL-18[(176.50±24.12)pg/ml与(182.50±20.51)pg/ml,P>0.05]浓度有所减低,但差异无统计学意义;草酸钙晶体刺激组较对照组细胞培养液中炎症因子IL-1β[(850.50±48.79)pg/ml与(183.50±7.78)pg/ml,P<0.001]和IL-18[(526.00±39.61)pg/ml与(182.50±20.51)pg/ml,P<0.001]浓度显著增高;ATV干预组较草酸钙晶体刺激组细胞培养液中炎症因子IL-1β[(452.50±36.06)pg/ml与(850.50±48.79)pg/ml,P<0.01]和IL-18[(403.50±23.33)pg/ml与(526.00±39.61)pg/ml,P<0.05]浓度显著减少。蛋白质印迹法检测结果显示,ATV组较对照组NF-κB的相对表达量有所减少(0.105±0.021与0.100±0.014),但差异无统计学意义(P>0.05);草酸钙晶体刺激组较对照组NF-κB的相对表达量明显增多(0.295±0.035与0.100±0.014,P<0.001);ATV干预组较草酸钙晶体刺激组NF-κB的相对表达量明显下降(0.160±0.012与0.295±0.035,P<0.05)。免疫荧光染色法检测NF-κB的表达趋势与蛋白质印迹法检测结果一致。结论草酸钙晶体能够诱导HK-2细胞的炎症反应,且ATV可抑制NLRP3炎症小体的活化,减少炎症因子IL-1β、IL-18的分泌和NF-κB的表达,发挥抗炎作用。

Investigation on high magnetoresistance Ni_(0.81)Fe_(0.19) films grown on (Ni_(0.81)Fe_(0.19))_(1-x)Cr_(x) underlayers

We have fabricated Ni0.81Fe0.19 films with (Ni0.81Fe0.19)1-xCrx films as underlayers by dc magnetron sputtering, the results show that larger anisotropic magnetoresistance (△R/R) values of Ni0.81Fe0.19 films are observed using the underlayers with Cr concentration of ~36 at.% at an optimum underlayer thickness of ~4.4 nm, the maximum AMR value is 3.35%. The results of atomic force microscope (AFM) and X-ray diffraction (XRD) show that the △R/R enhancement is attributed to the formation of large average grain size and the strong(111) texture in the Ni0.81Fe0.19 films.