清润方对糖尿病大鼠肝脏SIRT/NF-κB信号通路的影响

Effecf of Qingrun Formula on SIRT/NF-κB signal pathway in liver of diabetic rats

目的研究清润方对2型糖尿病胰岛素抵抗大鼠肝脏内沉默调节蛋白(SIRT)/核因子κB(NF-κB)信号通路的影响。方法选取90只12周龄的Wistar雄性大鼠作为研究对象,其中15只作为正常组,其余75只采用脂肪乳灌胃30 d和腹腔注射链脲佐菌素(STZ)的方法建立2型糖尿病胰岛素抵抗模型。将造模成功的60只糖尿病大鼠按照随机数字表法随机分成模型组、二甲双胍组[250 mg/(kg·d)]、清润方大剂量组[11.2 g/(kg·d)]、清润方中剂量组[5.6 g/(kg·d)]和清润方小剂量组[2.8 g/(kg·d)],每组12只。各组均灌胃给药,每天1次,连续8周。酶联免疫吸附测定(ELISA)法检测各组肝组织中肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的含量,q RT-PCR法和Western blot法检测肝组织中SIRT1、NF-κB和葡萄糖转运蛋白4(GLUT4)的m RNA和蛋白表达水平。结果与模型组比较,清润方大剂量组TNF-α水平降低(P<0.05),清润方大、中剂量组IL-6水平降低(P<0.05),清润方各剂量组SIRT1的m RNA和蛋白表达水平升高(P<0.05),清润方大、中剂量组GLUT4的m RNA和蛋白表达水平升高(P<0.05),清润方大、中剂量组NF-κB的m RNA和蛋白表达水平降低(P<0.05)。结论清润方可能通过促进糖尿病大鼠肝脏中SIRT1的表达,从而下调NF-κB的表达水平来改善胰岛素抵抗。

Objective To study the effect of Qingmn Formula on sirtuin （SIRT）/nuclear factor-κB （NF-κB） signal path- way in livers of rats with diabetes mellitus and insulin resistance. Methods Ninety 12-week old Wistar male rats were selected as objects, 15 rats were as normal group, the other 75 rats were induced by intraperitoneal injection of Strepto- zotocin （STZ） and gavage with fat milk for 30 days. After successful modeling, 60 rats with diabetes were randomly di- vided into model group, Metformin group [250 mg/（kg·d）], high dosage of Qingrun Formula group [11.2 g/（kg·d）], mid- dle dosage of Qingrun Formula group [5.6 g/（kg·d）] and low dosage of Qingrun Formula group [2.8 g/（kg·d）], with 12 rats in each group. Each group was given medicine by gavage, once a day, lasting for 8 weeks. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in liver tissues of each group were tested by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expression levels of SIRT1, NF-KB and glucose transporter 4 （GLUT4） in liver tissues of each group were tested by qRT-PCR and Western blot. Results Compared with model group, the level of TNF-α decreased in high dosage of Qingrun Formula group （P 〈 0.05）, the level of IL-6 decreased in high and middle dosage of Qingrun Formula groups （P 〈 0.05）, the mRNA and protein expression levels of SIRT1 increased in each dosage of Qingrun Formula groups （P 〈 0.05）, the mRNA and protein expression levels of GLUT4 increased in high and middle dosage of Qingrun Formula groups （P 〈 0.05）, the mRNA and protein expression levels of NF-κB decreased in the high and middle dosage of Qingrun Formula groups （P 〈 0.05）. Conclusion Qingrun Formula may alleviate the in- sulin resistance by increasing the expression of SIRT1 and thereby decreasing the expression of NF-κB in the liver of diabetic rats.