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隔药饼灸抑制TLR4延迟动脉粥样硬化形成机制的研究

The Research on Mechanism of Inhabitting Toll-like Receptor4 to Retard the Formation of Atherosclerosis Through Herbal-cake-separated-moxibustion

【作者】 何新群

【导师】 岳增辉;

【作者基本信息】 湖南中医药大学 , 针灸推拿学, 2010, 硕士

【摘要】 目的:观察隔药饼灸对动脉粥样硬化兔(atherosclerosis, AS)主动脉TLR4表达的影响,初步探讨隔药饼灸抑制TLR4延迟AS形成的作用机理。方法:将75只兔随机分为5组,每组15只,即:空白组、模型组、直接灸组、隔药饼灸组、药物组,采用高脂饲养加免疫损伤方法,建立AS模型,同时分别给予不同干预措施。实验进行16周后,取动脉血测定TC、TG、HDL-C、LDL-C、apo-A、apo-B含量;透射电镜观察兔主动脉内皮细胞超微结构;免疫组化检测TLR4的灰度值,观察其抗原免疫活性;荧光定量PCR测定TLR4mRNA表达。结果:①直接灸组、隔药饼灸组与药物组均能显著降低TC、TG含量(P<0.05或P<0.01),隔药饼灸组疗效优于直接灸组,而与药物组疗效相当;直接灸组、隔药饼灸组与药物组均可调整LDL、H/L值(P<0.05或P<0.01),隔药饼灸组与药物组作用相当(P>0.05);隔药饼灸组对apo-A、apo-B、A/B值调整作用与药物组相似(P>0.05)。②透射电镜观察主动脉内皮细胞分析显示:空白组内皮结构正常。模型组内皮层破坏严重,内含大量的脂滴和脂质,平滑肌细胞增生,排列紊乱,胞浆内充满脂肪空泡,主动脉呈典型AS改变。隔药饼灸组、药物组内皮细胞、内弹力膜结构基本正常,平滑肌细胞少量泡沫样变。直接灸组内皮细胞线粒体水肿、空泡样变,内皮层有少量泡沫细胞。③直接灸组、隔药饼灸组和药物组均使动脉粥样硬化兔主动脉内皮细胞TLR4及TLR4mRNA表达显著下调(P<0.05)结论:①隔药饼灸能有效降低AS兔血中TC、TG含量,且能降低血中LDL和apo-B含量,升高HDL和apo-A含量,提示隔药饼灸对脂质及脂蛋白的代谢具有良好的调整作用。②隔药饼灸对AS兔主动脉内皮细胞有一定的保护作用。③隔药饼灸能抑制主动脉内皮细胞TLR4活性,抑制主动脉TLR4mRNA表达,提示隔药饼灸是通过抑制TLR4表达从而延缓AS的形成。

【Abstract】 Objective:To observe the effect on Toll-like receptor 4(TLR4)mRNA of atherosclerosis rabbits treated by herbal-cake-separated moxibustion and to analyze the possible mechanism.Method:75 rabbits were divided into 5 groups, grouping follows: blank group、model group of AS、direct-moxibustion group、herbal-cake-separated moxibustion group and drug treatment group,15rabbits each group.While fed with high fat food everyday and immune injury except blank group.All these rabbits were feeding for 16weeks. Testing indicators of lipid,the change of ultrastructrre of endothelial cells of the aorta of the rabbits is observed with electron microscope,the expression of TLR4 mRNA.Results:①Compared with model group, the TC and TG content in herbal-cake-separated-moxibustion group、direct-moxibustion group and drug treatment group is obviously lower (P<0.05orP<0.01), Compared with direct-moxibustion group, TC content in herbal-cake-separated-moxibustion group is obviously lower(P<0.01). direct-moxibustion group. herbal-cake-separated moxibustion group and drug treatment group. The LDL content and the ratio of HDL/LDL in three treatment groups is lower than model group(P<0.05 or P<0.01); Herbal-cake-separated-moxibustion group and drug treatment group have the same efficacy (P>0.05); Herbal-cake-separated-moxibustion and drug treatment group have similar regulating effect to the content of apo-A. apo-B, the ratio of A/B (P>0.05).②Under transmission electron microscope, observation of aortic endothelial cells shows:endothelial cells in direct-moxibustion group and herbal-cake-separated-moxibustion group are intacter, most of them are normal in forms, internal elastic membrane is normal, the cells of muscles of medial membrane are relatively normal and the structure of aortic endothelial cells of those groups are similar to those of blank group.The structure of aortic endothelial cells in model group is AS typical lesions, endothelial cells thanatosis, in cytoplasm the sedimentation of a great quantity of lipide can be seen, internal elastic membrane is disruption severely, there is focal dissolution, the cells of smooth muscle in the medial membrane disarragement, and has seriously frothy change.③The expression of TLR4 and TLR4mRNA.in direct-moxibustion group. herbal-cake-separated moxibustion group and drug treatment group is obviously decreased(P<0.05).Conclusion:①Satisfied efficacy on metabolism of lipid、lipoprotein of AS rabbits after treated by direct-moxibustion、herbal-cake-separated moxibustion and drug treatment, herbal-cake-separated moxibustion and drug treatment have the similar effects,both of these method are better than direct-moxibustion.②herbal-cake-separated moxibustion、direct-moxibustion and drug treatment have protective effects on AS rabbit aortic endothelial cells on some contents.③The herbal-cake-separated moxibustion have a certain extent inhibition to the expression of the aortic endothelial cells TLR4.These shows the herbal-cake-separated moxibustion through inhabitting Toll-like receptor4 to retard the formation of Atherosclerosis.

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