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目的探讨高脂饮食喂养对肥胖大鼠脂肪组织中TLR4炎症信号通路的影响。方法将20只SPF级雄性4周龄SD大鼠随机分为正常饮食组(NC,n=10)和高脂饮食组(HFD,n=10)。检测每组大鼠空腹血糖及胰岛素水平,并计算HOMA-IR;应用Real-time PCR及Western blot方法检测脂肪组织中炎症信号通路TLR4、TRAF6、IKKβ、TNF-α、IL-6的表达。结果 (1)与NC组相比,HFD组空腹胰岛素水平显著升高[分别为(14.64±0.66)mIU/L vs.(31.71±3.61)mIU/L,P=0.005];HOMA-IR值升高(分别为3.40±0.15 vs.7.59±0.99,P=0.008);两组大鼠空腹血糖水平无明显统计学差异。(2)Real time PCR结果显示,HFD组脂肪组织中TLR4、TRAF6、TNF-α及IL-6 mRNA水平均明显升高,分别为NC组的1.85倍(P=0.002)、1.72倍(P=0.031)、3.82倍(P=0.045)及3.17倍(P=0.003)。(3)HFD组大鼠炎症因子TLR4、IKKβ及TNF-α蛋白水平亦明显升高,分别为NC组的1.62倍(P=0.000)、1.77倍(P=0.003)及3.02倍(P=0.002)。结论高脂饮食可通过介导TLR4炎症信号通路活化,使脂肪组织中炎症因子TNF-α、IL-6表达升高,进而导致胰岛素抵抗的发生。
Objective To investigate the effect of high-fat diet on TLR4 signaling pathway in adipose tissue of obese rats. Methods Twenty male SPF rats of 4 weeks old were randomly divided into normal diet group (NC, n = 10) and high fat diet group (HFD, n = 10). Fasting plasma glucose and insulin levels were measured in each group, and HOMA-IR was calculated. Real-time PCR and Western blot were used to detect the expression of TLR4, TRAF6, IKKβ, TNF-α and IL-6 in adipose tissue. Results (1) Compared with NC group, the fasting insulin level in HFD group was significantly higher than that in NC group [(14.64 ± 0.66) mIU / L vs. (31.71 ± 3.61) mIU / L, P = 0.005] (3.40 ± 0.15 vs.7.59 ± 0.99, P = 0.008, respectively). There was no significant difference in fasting plasma glucose between the two groups. (2) The results of Real time PCR showed that the levels of TLR4, TRAF6, TNF-α and IL-6 mRNA in adipose tissue of HFD group were significantly higher than that of NC group (1.85-fold and 1.72- 0.031), 3.82-fold (P = 0.045) and 3.17-fold (P = 0.003). (3) The protein levels of TLR4, IKKβ and TNF-αin HFD group were also significantly higher than those in NC group (1.62-fold, 1.77-fold and 1.30- ). Conclusion High-fat diet can activate TLR4 signaling pathways and increase the expression of inflammatory cytokines TNF-α and IL-6 in adipose tissue, leading to the development of insulin resistance.