百合乌药汤对1型糖尿病并发肝损伤的保护作用及其机制分析

Protective Effect and Its Mechanism Analysis of Baihe Wuyao Decoction on Treatment of Type 1 Diabetes Mellitus and Associated Hepatic Injury

目的:考察百合乌药汤(Baihe Wuyao decoction,BWD)对小鼠1型糖尿病(Type 1 diabetes mellitus,T1DM)并发肝损伤的保护作用,并揭示其潜在机制。方法:小鼠70只,随机选取10只作为空白组,其余采用腹腔注射链脲佐菌素(Streptozotocin,STZ)诱导T1DM,成模后随机分为6组:模型组、阳性对照组、BWD各剂量组(15、 5、 2.5、 1.25 g·kg^(-1)·d^(-1))。各组小鼠连续给药6周。检测小鼠血清中谷丙转氨酶(Alanine aminotransferase,ALT)、谷草转氨酶(Aspartate aminotransferase,AST)的含量,及肝组织匀浆超氧化物歧化酶(Superoxidedismutase,SOD)和丙二醛(Malonaldehyde,MDA)的含量;观察小鼠肝脏病理学变化;检测小鼠肝脏中蛋白激酶B(Protein kinase B,AKT)在蛋白水平的表达,检测锰超氧化物歧化酶(Manganese superoxide dismutase,Mn-SOD)和一氧化氮合酶(Inducible nitric oxide synthase,iNOS)、炎症因子包括肿瘤坏死因子(Tumor necrosis factor,TNF-α)、核因子-κB(Nuclear factor-κB,NF-κB)、白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)和白细胞介素-8(Interleukin-8,IL-8)、凋亡因子包括(B-cell lymphoma-2,Bcl2)、(Bcl2-associated X protein,Bax)和(Caspase 3,CASP3)在m RNA水平的表达。结果:BWD可改善T1DM小鼠肝组织病理损伤;与模型组相比,各剂量组ALT和AST水平均显著降低(P<0.01或P<0.05),低1剂量组效果更明显;BWD低1剂量可显著上调p-AKT/AKT(P<0.05);低1剂量显著下调MDA、iNOS,上调SOD、Mn-SOD含量(P<0.01或P<0.05);与模型组相比,低1剂量的Bcl2/Bax比值明显增加,Bax、CASP3被下调;低1剂量显著降低TNF-α、NF-κB、IL-1β、IL-6和IL-8在基因水平的表达(P<0.01或P<0.05)。结论:BWD可通过抗炎、抗氧化、抗凋亡、促进细胞增殖以及改善胰岛素信号通路发挥对T1DM并发肝损伤的保护作用。

Objective: To investigate the protective effect of Baihe Wuyao decoction(BWD) on mice with type 1 diabetes mellitus(T1 DM) and liver damage, and to reveal its underlying mechanism. Methods: Ten of 70 mice were randomly selected as the blank group, and the rest were induced with intra-peritoneally(ip) streptozotocin(STZ) to induce T1 DM.After successful modeling, the mice were randomly divided into 6 groups: Model group, positive control group and BWD groups(15, 5, 2.5, 1.25 g·kg^(-1)·d^(-1)). The mice in each group were given continuous administration for 6 weeks. The contents of alanine aminotransferase(ALT) and aspartate aminotransferase(AST) in the serum of mice were measured, and the contents of superoxidedismutase(SOD) and malonaldehyde(MDA) in liver tissue homogenate were detected. The pathological changes of mouse liver were observed. The expression of Protein kinase B(AKT) in mouse liver was detected,the mRNA level of manganese superoxide dismutase(Mn-SOD) and nitric oxide synthase(iNOS), inflammatory factors including tumor necrosis factor(TNF-α), nuclear factor-κB(NF-κB), interleukin-1β(IL-1β), interleukin-6(IL-6),interleukin-8(IL-8), apoptosis factors including B-cell lymphoma-2(Bcl2), Bcl2-associated X protein(Bax), Caspase 3(CASP3) were measured. Results: BWD exhibited a good hepatic protection with ameliorating hepatic pathological morphology in T1 DM mice. Compared with the model group, the levels of ALT and AST in each dose group were significantly reduced(P<0.01, P<0.05), and the effect of the low 1 dose group was more obvious. The low 1 dose of BWD could significantly up-regulate p-AKT/AKT(P<0.05), low dose 1 could significantly down-regulate MDA, iNOS, and upregulate SOD and Mn-SOD content(P<0.01, P<0.05). Comparing with the model group, the ratio of Bcl2/Bax at the low dose increased significantly, Bax and CASP3 were down-regulated, the low dose significantly reduced the expression of TNF-α, NF-κB, IL-1β, IL-6 and IL-8 at the gene level(P<0.01, P<0.05). Conclusion: The BWD would have a good hepatic protective effect during T1 DM, which underlying mechanisms through improving insulin resistance and suppresses oxidative stress, apoptosis, inflammation and promotion of cell proliferation.