| 朱博,陈攀,王辉,丁科.理气化痰祛瘀方对高脂血症金黄地鼠炎症的干预作用及机制研究[J].浙江中西医结合杂志,2020,30(9): |
| 理气化痰祛瘀方对高脂血症金黄地鼠炎症的干预作用及机制研究 |
| Study on the effect and mechanism of Liqi Huatan Quyu Decoction on the inflammation of golden hamster with hyperlipidemia |
| 投稿时间：2020-04-14 修订日期：2020-07-31 |
| DOI： |
| 中文关键词: 理气化痰祛瘀方 高脂血症 炎症 TLR4/TRAF-6 |
| 英文关键词:Liqi Huatan Quyu decoction hyperlipidemia TLR4/TRAF-6 |
| 基金项目:浙江省中医药科技计划项目(2017ZB042) |
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| 中文摘要: |
| 目的 观察理气化痰祛瘀方对高脂血症金黄地鼠炎症的表达水平，初步研究TLR4/ TRAF-6信号通路在其中所起的作用。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对照组（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6.3，12.6，25.2 g·kg-1·d-1）和非诺贝特组（150mg·kg-1·d-1）进行治疗，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）含量、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, qRT-PCR检测金黄地鼠肝脏组织中TLR4、TRAF-6 mRNA水平表达情况。 结果 理气化痰祛瘀方能显著降低金黄地鼠血清TC、TG、LDL-c水平，与模型组比较，对照组中理气化痰祛瘀方中、高剂量组金黄地鼠血清IL-6显著下调[（128.65±16.73）pg/mL比（145.96±11.62）pg/mL；（124.48±20.48）pg/mL比（145.96±11.62)pg/mL P均<0.05]；理气化痰祛瘀方中、高剂量组金黄地鼠血清TNF-α含量显著下调[（447.73±65.28）ng/L比（508.92±20.13）ng/L;（442.01±53.19）ng/L比（508.92±20.13)ng/L ，P均<0.05]；理气化痰祛瘀方中、高剂量组肝脏 CYP7A1含量则显著上升[（32.43±6.08）ng/mL比（25.85±2.49）ng/mL，P<0.01；（33.70±5.74）ng/mL比（25.85±2.49)ng/mL, 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蛋白表达水平[（1.651±0.188）比（2.158±0.124），P<0.05；（1.411±0.134）比（2.158±0.124)，P<0.01；（1.346±0.188）比（2.158±0.124)，P<0.01]和TRAF-6蛋白表达水平[（1.785±0.048）比（2.283±0.216），P<0.05；（1.605±0.138）比（2.283±0.216)，P<0.01； （1.442±0.209）比（2.283±0.216)，P<0.01]均显著下调，且呈现剂量依赖性；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中TLR4 mRNA水平[（56.32±15.56）g/kg比（122.90±27.94）g/kg，P<0.05；（20.83±4.34）g/kg比（122.90±27.94）g/kg，P<0.01；（9.88±2.50）g/kg比（122.90±27.94）g/kg ，P<0.01]均显著下调，TRAF-6 mRNA水平[（23.62±3.85）g/kg比（55.79±4.56）g/kg；（11.23±2.46）g/kg比（55.79±4.56）g/kg；（6.29±2.15）g/kg比（55.79±4.56）g/kg ，P均<0.01]均显著下调，且呈现剂量依赖性。结论 理气化痰祛瘀方参与高脂饲料诱导的金黄地鼠高脂血症中炎症的发生，显著抑制IL-6、 TNF-α、 CYP7A1在高脂血症金黄地鼠血清及肝脏中的表达，其机制可能与TLR4/ TRAF-6 信号通路传导途径相关。 |
| 英文摘要: |
| Objective To observe the expression level of liqihuatu quyu decoction on hyperlipidemia golden hamster inflammation and the role of TLR4 / TRAF-6 signaling pathway in it. Methods The golden hamster model of hyperlipidemia was induced by high-fat feed. Forty-eight seven-week-old male LVG Syrian golden hamsters were randomly divided into the normal control group (0.9% normal saline) by using a high-fat diet-induced hyperlipidemia golden hamsters model, low, middle and high dose of Liqi Huatan quyu Fang (6.3，12.6，25.2 g·kg?1·d-1) and non-Norbert group (150mg·kg-1·d-1) were treated respectively. The blood biochemical indexes were measured 6 weeks after administvsn (8 weeks after feeding) .The serum IL-6、TNF-α levels and liver tissue CYP7A1 levels were detected by ELISA.The expression of TLR4 and TRAF-6 protein in the liver tissue of golden hamster was detected by Western blot, and the expression of TLR4 and TRAF-6 mRNA in the liver tissue of golden hamster was detected by qRT-PCR. Result The intervention of Liqi Huatan Quyu decoction on hyperlipidemia golden hamster can significantly reduce the TC、TG、LDL-c level and serum IL - 6, TNF-α content. Compared with the model group, the serum IL-6 of Golden Hamsters in the middle and high dose groups decreased significantly[（128.65±16.73）pg/mL vs（145.96±11.62）pg/mL；（124.48±20.48）pg/mL vs（145.96±11.62)pg/mL P<0.05].The expression of TNF-αprotein in the liver of golden hamster was significantly decreased in the middle and high dose groups of Liqi Huatan Quyu recipe[（447.73±65.28）ng/L vs（508.92±20.13）ng/L;（442.01±53.19）ng/L vs（508.92±20.13)ng/L ，P<0.05] ,and the expression of CYP7A1 was significantly reduce in the two groups[（32.43±6.08）ng/mLvs（25.85±2.49）ng/mL，P<0.01；（33.70±5.74）ng/mL vs（25.85±2.49)ng/mL, P<0.05] .Compared with model group, TLR4 [（1.651±0.188）vs（2.158±0.124），P<0.05；（1.411±0.134）vs（2.158±0.124)，P<0.01；（1.346±0.188）vs（2.158±0.124)，P<0.01]and TRAF-6 [（1.785±0.048）vs（2.283±0.216），P<0.05；（1.605±0.138）比（2.283±0.216)，P<0.01； （1.442±0.209）vs（2.283±0.216)，P<0.01] protein expression in liver tissue of Golden Hamsters in low, middle and high doses of Liqi Huatan quyu prescription were significantly decreased, and showed dose-dependent the expression of TLR4[（56.32±15.56）g/kg vs（122.90±27.94）g/kg，P<0.05；（20.83±4.34）g/kg vs（122.90±27.94）g/kg，P<0.01；（9.88±2.50）g/kg vs（122.90±27.94）g/kg ，P<0.01] and TRAF-6 mRNA [（23.62±3.85）g/kg vs（55.79±4.56）g/kg,（11.23±2.46）g/kg vs（55.79±4.56）g/kg，（6.29±2.15）g/kg vs（55.79±4.56）g/kg ，P<0.01] gene was significantly decreased in the two groups . Conclusion Liqi Huatan Quyu decoction is involved in the occurrence of inflammation in hyperlipidemia golden hamster induced by high-fat diet. It can significantly inhibit the expression of IL-6, TNF - α, CYP7A1 in serum and liver of golden hamster with hyperlipidemia. Its mechanism may be related to TLR4 / TRAF-6 signaling pathway. |
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