苏子油对大鼠体内细胞因子及肾草酸钙结晶形成影响的研究

Effect of perilla frutescens(L·) Britt·oil on cytokines and renal calcium oxalate crystallization formation in rats

目的了解炎性细胞因子白细胞介素-1α(IL-1α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)在尿结石形成中的作用以及α-亚麻酸(-αL inolen ic ac id,α-LNA)对这些因子及肾草酸钙结晶形成的影响。方法45只雄性W istar成年大鼠随机分成空白对照组(Ⅰ)、成石组(Ⅱ)和苏子油组(Ⅲ),分别接受普通饲料加自来水饮水,普通饲料加诱石剂饮水和普通饲料加诱石剂饮水并每日苏子油灌胃(2 g.只-1.d-1),实验8周后检测各组大鼠肾功能,24 h血尿生化指标,肾草酸钙结晶情况及IL-1α、IL-1β、IL-6和TNF-α水平。利用体外培养的大鼠肾系膜细胞(RMC),设立空白对照组、10μg/L IL-6处理组、EPA 10μmol/L+IL-6 10μg/L处理组、EPA 50μmol/L+IL-6 10μg/L处理组以及无水乙醇处理组,采用放免法检测培养上清中PGE2含量。结果苏子油组肾组织水肿较轻,肾内草酸钙结晶数及肾钙含量明显低于成石组(P<0.01),24 h尿钙排泄,血尿素氮,肌酐浓度显著低于成石组(P<0.05),苏子油组和对照组IL-1α、IL-6水平显著低于成石组(P<0.01),3组间IL-1β、TNF-α水平差异无显著性(P>0.05)。细胞培养上清PGE2含量,空白对照组、EPA 10μmol/L+IL-6 10μg/L处理组、EPA 50μmol/L+IL-6 10μg/L处理组显著低于10μg/L IL-6处理组(P<0.01),EPA 50μmol/L+IL-6 10μg/L处理组显著低于EPA 10μmol/L+IL-6 10μg/L处理组(P<0.01),空白对照组、无水乙醇处理组二组间差异无显著性(P>0.05)。结论PGE2α-亚麻酸可能通过抑制炎性细胞炎子的产生而对尿石形成起保护作用,其在尿石症防治方面可能有一定的应用价值。

Objective To investigate the possible role of cytokines interleukin （IL）-1β, IL-1α, IL-6 and TNF-α in urinary stone formation and the inhibitory effect of α-Linolenic acid on these cytokines and renal calcium oxalate crystallization formation in rats. Methods Forty-five male adult Wistar rats were divided receive normal forage and water as negative control （ Ⅰ ）, normal forage and water containing 1% ethylene glycol and 1% muriat to induce renal stone as group Ⅱ , or normal forage and water containing 1% ethylene glycol and 1% muriat and 10 g · kg^-1 · d^-1 perilla frutescens （L · ） Britt · oil as group Ⅲ for 8 weeks. Then all rats were sacrificed and the renal tissue calcium, the urinary calcium, the calcium oxalate crystallization and urinary oxalate and serum IL-1β, IL-α, IL-6 and TNF-α were detected. RMCs were pretreated with 10 μg/L IL-6, 10 μmol/L EPA ＋ 10 μg/L IL-6, 50 μmol/L EPA ＋ 10 μg/L IL-6, or absolute alcohol for 24 h. RMCs without pretreatment served as normal control. The PGE2 concentration in RMC supernatant was measured by radioimmunoassay. Results As compared with group Ⅱ , the hydronephrosis and parenchymal swelling were obviously slight, and calcium oxalate crystalization, renal tissue calcium, urinary calcium excretion, blood urea nitrogen and creatinine were significantly lower in group Ⅲ （P 〈 0.05 ）. The serum level of IL-α, IL-6 in groupsⅠ and Ⅲ was significantly lower than group Ⅱ （ P 〈 0. 01 ） , while the level of IL-1β and TNF-α showed no significant difference among the 3 groups. The PGE2 concentration in RMC supernatant were significantly lower in groups A, C and D than that of group B （P〈0. 01 ） , and lower in group D than group C （P〈0. 01 ） , but of no difference between group A and E. Conclusion α-Linolenic acid may prevent urinary stone formation by the inhibitory role on these cytokines.