至宝三鞭丸基于Nrf2/HO-1通路的抗疲劳作用机制研究

Study on the Anti-fatigue Mechanism of Zhibao Sanbian Pills Based on Nrf2/HO-1 Pathway

目的通过观察至宝三鞭丸(Zhibao Sanbian Pills)对Nrf2/HO-1信号通路的影响,研究其对小鼠的抗疲劳作用及机制。方法将ICR小鼠随机分为正常对照组、至宝三鞭丸低剂量(0.958 g·kg^(-1))组、至宝三鞭丸中剂量(4.79 g·kg^(-1))组、至宝三鞭丸高剂量(9.58 g·kg^(-1))组,每组25只,连续给药35 d,记录体质量。采用负重游泳实验评价至宝三鞭丸的抗疲劳作用;比色法检测各组小鼠血清中的尿素氮(Urea nitrogen,UN)和乳酸(Lactic acid)水平;比色法检测各组小鼠肝组织中的糖原(Liver glycogen)含量;Western Blot检测各组小鼠肝组织中核因子红细胞2相关因子2(Nuclear factor erythroid 2-related factor 2,Nrf2)、核孔糖蛋白62(Sequestosome 1,P62/SQSTM1)、血红素加氧酶1(Heme oxygenase-1,HO-1)和醌氧化还原酶1(Quinone oxidoreductase 1,NQO1)的蛋白表达水平。结果与正常对照组比,至宝三鞭丸各剂量组可不同程度延长小鼠负重游泳时间,提高肝脏组织中糖原含量,下调血清中尿素氮水平,其中至宝三鞭丸高剂量(9.58 g·kg^(-1))组的差异具有统计学意义(均P<0.05);至宝三鞭丸虽具有降低乳酸堆积的作用趋势,但差异无统计学意义(P>0.05);此外,至宝三鞭丸各剂量组均可提高肝组织中Nrf2、P62、HO-1和NQO1的蛋白表达水平(P<0.05,P<0.01)。结论至宝三鞭丸具有明显的抗疲劳作用,其机制可能与激活Nrf2/HO-1信号通路,从而提高机体的抗氧化能力有关。

Objective To study the anti-fatigue effect and mechanism of Zhibao Sanbian Pills(ZB) on mice by observing its effect on Nrf2/HO-1 signaling pathway.Methods The ICR mice were randomly divided into four groups,including the control group,ZB low-dose(0.958 g·kg^(-1))group,ZB medium-dose(4.79 g·kg^(-1))group and ZB high-dose(9.58 g · kg^(-1)) group, 25 mice in each group, then they were intragastrically administered for35 days, and the body mass were recorded. Weight-bearing swimming test was used to evaluate the antifatigue effect of ZB. Colorimetry was used to detect urea nitrogen(UN)and lactic acid(LA)level in the serum of each group, as well as the content of liver glycogen(LG) in liver tissue. The protein expressions of nuclear factor erythroid 2-related factor 2(Nrf2), sequestosome 1(P62/SQSTM1), heme oxygenase-1(HO-1)and quinone oxidoreductase 1(NQO-1) in liver tissue were detected by Western Blot.Results Compared with the control group,each dose group of ZB can prolong the exhaustive swimming time of mice in different degree,increase the content of LG in liver tissue, and decrease the content of serum UN. ZB high-dose group(9.58 g · kg^(-1)) had significant statistical difference(P<0.05). ZB can reduce lactic acid accumulation, but there is no statistical difference(P>0.05). In addition,the protein expression levels of Nrf2,P62,HO-1 and NQO-1 in liver tissue were increased in all ZB administration groups(P<0.05,P<0.01).Conclusion ZB plays an obvious anti-fatigue role by regulating Nrf2/HO-1 signaling pathway to improve antioxidant capacity of the body.