理气化痰祛瘀方对高脂血症模型金黄地鼠炎症的干预作用及机制研究

Intervention Effect and Mechanism of Liqihuatanquyu Recipe on Inflammation in Hyperlipidemia Golden Hamster

目的探讨理气化痰祛瘀方对高脂血症模型金黄地鼠炎症的干预作用及其作用机制。方法采用高脂饲料喂养诱导高脂血症金黄地鼠模型,按数字表随机分配法将48只7周龄雄性LVG叙利亚金黄地鼠随机分为正常组和模型组(等量0.9%生理盐水),理气化痰祛瘀方低、中、高剂量组(剂量依次为6.3、12.6、25.2g·kg-1·d-1)和非诺贝特组(150mg·kg^(-1)·d^(-1)),每组各8只。给药后6周(饲喂第8周)测定血生化指标。采用酶联免疫吸附(ELISA)法分别检测血清白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)及肝脏组织胆固醇7α羟化酶(CYP7A1)含量。Western blot检测给药处理后金黄地鼠肝脏组织Toll样受体4(TLR4)和肿瘤坏死因子受体相关蛋白6(TRAF-6)水平表达情况,q RT-PCR检测金黄地鼠肝脏组织TLR4、TRAF-6 m RNA水平表达情况。结果与模型组比较,理气化痰祛瘀方中、高剂量组金黄地鼠血清TC[(20.74±7.24)mmol/L、(22.38±7.03)mmol/L比(38.61±10.67)mmol/L,P<0.05]、TG[(5.14±1.68)mmol/L、(5.79±2.80)mmol/L比(11.80±4.89)mmol/L,P<0.05]、LDL-c[(8.56±3.54)mmol/L、(8.76±3.07)mmol/L比(15.84±5.98)mmol/L,P<0.05]水平下降,且理气化痰祛瘀方中剂量组下降更明显(P<0.05);与模型组比较,理气化痰祛瘀方中、高剂量组金黄地鼠血清IL-6[(128.65±16.73)pg/m L、(124.48±20.48)pg/m L比(145.96±11.62)pg/m L,P均<0.05]、TNF-α[(447.73±65.28)ng/L、(442.01±53.19)ng/L比(508.92±20.13)ng/L,P均<0.05]含量显著下调,肝脏组织CYP7A1含量则显著上升[(32.43±6.08)ng/m L、(33.70±5.74)ng/m L比(25.85±2.49)ng/m L,P均<0.05],且呈剂量依赖性(P<0.05);与模型组比较,理气化痰祛瘀方低、中、高剂量组金黄地鼠肝脏组织TLR4[(1.651±0.188)、(1.411±0.134)、(1.346±0.188)比(2.158±0.124),P<0.05]和TRAF-6[(1.785±0.048)、(1.605±0.138)、(1.442±0.209)比(2.283±0.216),P均<0.05]蛋白表达水平均显著下调,且呈现剂量依赖性(P<0.05);与模型组比较,理气化痰祛瘀方低、中、高剂量组金黄地鼠肝脏组织TLR4 m RNA[(56.32±15.56)、(20.83±4.34)、(9.88±2.50)比(122.90±27.94),P均<0.05]及TRAF-6m RNA水平[(23.62±3.85)、(11.23±2.46)、(6.29±2.15)比(55.79±4.56),P均<0.05]均显著下调,且呈现剂量依赖性(P<0.05)。结论理气化痰祛瘀方对高脂饲料诱导的高脂血症模型金黄地鼠炎症因子有干预作用,显著抑制IL-6、TNF-α、CYP7A1在高脂血症模型金黄地鼠血清及肝脏表达,其机制可能与TLR4/TRAF-6信号通路传导途径相关。

Objective To explore the intervention effect and mechanism of Liqihuatanquyu Recipe on inflammation of golden hamster with hyperlipidemia model.Methods The hyperlipidemia golden hamster model was induced by feeding high-fat diet.Forty eight 7-week-old male LVG Syrian golden hamsters were randomly divided into normal group and model group(equivalent 0.9%saline),low-,medium-and high-dose Liqihuatanquyu Recipe groups(6.3,12.6 and 25.2g·kg-1·d-1,respectively),and fenofibrate group(150mg·kg-1·d-1),with 8 animals in each group.The blood biochemical indexes were measured 6 weeks after treatment(feeding week 8).Enzyme-linked immunosorbent assay(ELISA)was used to detect the contents of serum interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and cholesterol 7-αhydroxylase(CYP7A1)in liver tissue.The expressions of toll-like receptor 4(TLR4)and tumor necrosis factor receptor-related protein 6(TRAF-6)in the liver tissue were tested by Western blot and qRT-PCR,respectively.Results Compared to the model group,the serum levels of TC[(20.74±7.24)and(22.3±7.03)vs(38.61±10.67)mmol/L,P<0.05],TG[(5.14±1.68)and(5.79±2.80)vs(11.80±4.89)mmol/L,P<0.05]and LDL-c[(8.56±3.54)and(8.76±3.07)vs(15.84±5.98)mmol/L,P<0.05]in medium-and high-dose Liqihuatanquyu Recipe groups decreased significantly,and medium-dose group reduced more significantly(P<0.05);The contents of IL-6[(128.65±16.73)and(124.48±20.48)vs(145.96±11.62)pg/mL,P<0.05]and TNF-α[(447.73±65.28)and(442.01±53.19)vs(508.92±20.13)ng/L,P<0.05]in the medium-and high-dose groups were significantly decreased,while the content of CYP7A1[(32.43±6.08)and(33.70±5.74)vs(25.85±2.49)ng/mL,P<0.05]was significantly increased,and in a dose-dependent manner(P<0.05);The protein expression levels of TLR4[(1.651±0.188),(1.411±0.134)and(1.346±0.188)vs(2.158±0.124),P<0.05]and TRAF-6[(1.785±0.048),(1.605±0.138)and(1.442±0.209)vs(2.283±0.216),P<0.05]in the liver tissues of golden hamsters in the low-,medium-and high-dose groups were significantly decreased,and in a dose-dependent manner(P<0.05);The mRNA levels of TLR4[(56.32±15.56),(20.83±4.34)and(9.88±2.50)vs(122.90±27.94)]and TRAF-6[(23.62±3.85),(11.23±2.46)and(6.29±2.15)vs(55.79±4.56),P<0.05]in the liver tissues of golden hamsters in the low-,medium-and high-dose groups were significantly decreased,and in a dose-dependent manner(P<0.05).Conclusion Liqihuatanquyu Recipe can significantly inhibit the expression of IL-6,TNF-αand CYP7A1 in serum and liver of hyperlipidemia golden hamster induced by high-fat diet.The mechanism may be related to TLR4/TRAF-6 signaling pathway.