艾灸及艾烟对ApoE^-/-小鼠动脉粥样硬化胆固醇代谢和炎性反应的影响

【作者】 崔莹雪；

【导师】 赵百孝；

【作者基本信息】 北京中医药大学 ， 针灸推拿， 2014， 博士

【摘要】 目的：观察艾灸及艾烟干预对动脉粥样硬化的影响,并从调节脂代谢、抑制炎症等角度探索艾灸及艾烟的作用机制。方法：51只8周龄ApoE-/-小鼠随机分为3组：模型组,艾烟组,艾灸组,每组17只,20只同龄C57BL/6小鼠作为正常对照。正常组、模型组小鼠常规抓取、固定；艾烟组小鼠暴露于10mg/m3-15mg/m3艾烟环境；艾灸组小鼠艾灸关元穴。所有干预20min/日,6日／周,连续干预12周。血生化法检测血清’TC、TG、HDL-C、LDL-C、VLDL-C、ox-LDL、ApoA-I含量；elisa法测定血清TNFα、IL-1、IL-6、IL-10、hs-CRP、vWF、ICAM-1、VCAM-1、MCP-1的含量；HE染色、油红“0”染色观察主动脉根、胸主动脉病理改变并定量分析斑块大小；western blot法检测主动脉组织中ABCA1、ABCG1、LXRα、CD36、PPARγ、NFKB的表达情况；HE染色观察肝脏病理形态；免疫组化法检测肝脏ABCA1、LXRα、CD36、 PPARγ表达。结果：1.血脂水平模型组小鼠血清TG、LDL、VLDL含量显著高于正常组(p=0.003,0.001,0.004)ApoA-I、HDL含量显著低于正常组(p=0.001,0.007),TC、ox-LDL含量与正常组差异不显著(p>0.05)；艾灸组小鼠血清TG、LDL含量显著低于模型组(p=0.03,0.001),ApoA-Ⅰ含量显著高于模型组(p=0.001),VLDL、HDL含量与模型组无显著差异(p=0.11,0.11)；艾烟组小鼠血清TG、LDL含量显著低于模型组(p=0.01,0.008),ApoA-Ⅰ含量显著高于模型组(p=0.01),VLDL、HDL含量与模型组无显著差异(p=0.24,0.11)。艾灸组与艾烟组各项血脂指标均无显著性差异(p>0.05)。2.血清炎性因子和主动脉NFKB表达模型组小鼠血清IL-1、IL-6、TNFα、hs-CRP、vWF、ICAM-1、VCAM-1、MCP-1含量显著高于正常组(p<0.001),IL-10含量显著低于正常组(p<0.001)；艾灸组小鼠血清IL-1、IL-6、TNFα、hs-CRP、vWF、ICAM-1、VCAM-1、MCP-1含量显著低于模型组(p=0.005,0.001,<0.001,0.01,0.001,0.001,<0.001,0.002),IL-10含量显著高于模型组(p<0.001)；艾烟组小鼠血清IL-1、IL-6、TNFα、hs-CRP、vWF、ICAM-1、VCAM-1、MCP-1含量显著低于模型组(p=0.003,0.003,<0.001,0.01,0.02,0.03,<0.001,0.005),IL-10含量显著高于模型组(p=0.01)；艾灸组小鼠各项指标与艾烟组无显著差异(p>0.05)。各组小鼠胸主动脉NFKB表达差异不显著。3.主动脉粥样硬化斑块面积比各组ApoE-／-小鼠主动脉根部斑块面积比分别为：模型组15.76%,艾烟组8.55%,艾灸组7.34%；胸主动脉斑块面积比分别为：模型组9.05%,艾烟组4.37%,艾灸组2.47%。艾灸组显著低于模型组(主动脉根p=0.04,胸主动脉p=0.01)；艾烟组与模型组差异不显著(主动脉根p=0.08,胸主动脉p=0.08)；艾灸组与艾烟组差异不显著(p>0.05)。4.主动脉ABCA1、ABCG1、CD36、LXRα、PPARγ表达模型组小鼠胸主动脉ABCA1、ABCG1、LXR α表达显著低于正常组(p<0.001,0.04,0.004);CD36、PPAR γ表达与正常组无显著性差异(p>0.05)；艾灸组小鼠胸主动脉ABCA1、LXRα表达显著高于模型组(p=0.05,0.03),ABCG1表达与模型组无显著性差异(p=0.14)；艾烟组ABCA1、LXR α表达显著高于模型组(p<0.001,0.02),ABCG1表达与模型组无显著性差异(p=0.14)；艾灸组与艾烟组ABCA1、ABCG1、LXR α表达无显著差异(p>0.05)。5.肝脏ABCA1、CD36、LXRα、PPARγ表达模型组小鼠肝脏ABCA1、PPARγ表达显著低于正常组(P<0.001,<0.001),CD36、LXR a表达显著高于正常组(p=0.004,0.002)；艾灸组ABCA1、 PPARγ表达显著高于模型组(p=0.03,0.03),LXR α表达显著低于模型组(p=0.03),CD36表达与模型组无显著性差异(p=0.09)；艾烟组ABCA1、PPARγ表达显著高于模型组(p=0.002,0.01),CD36、LXRα表达显著低于模型组(p=0.02,0.02)；艾灸组与艾烟组ABCA1、CD36、LXR α、PPARγ表达均无显著差异(p>0.05)结论：1.艾灸可以有效缓解动脉粥样硬化病变,抑制动脉粥样硬化斑块生长；艾烟有抑制动脉粥样硬化斑块生长的作用趋势。2.艾灸和艾烟可以调节血脂代谢、促进病变处胆固醇外流、改善肝脏脂代谢功能,是防治动脉粥样硬化的机制之一。3.艾灸和艾烟可以抑制体内炎性反应,是防治动脉粥样硬化的机制之一。更多还原

【Abstract】 Objective:To observe the effect of moxibustion and moxa smoke on atherosclerosis and to explain their effect mechanism on lipid metablosim and inflammation.Methods:Sixty-eight ApoE-/-mice (8weeks old) were randomly divided into3groups (17in each group):model control group, moxa smoke group, moxibustion group, twenty same age C57BL/6mice were used as blank controls. Mice in the blank control and model control group were handled everyday. Mice in the moxa smoke group were exposed to moxa smoke with a concentration of10-15mg/m3while mice in the moxibustion group were treated with moxibustion on GuanYuan (RN4) point. All the manipulations were did20min each day,6days each week,12weeks totally.Blood biochemistry method was used to test the level of TC, TG, HDL-C, LDL-C, VLDL-C, ox-LDL, ApoA-Ⅰ in the serum. Elisa method was used to test the level of TNFa, IL-1, BL-6, IL-10, hs-CRP, vWF, ICAM-1, VCAM-1, MCP-1in the serum. HE and Red oil "O" stain were used to observe the plague area in the aorta root and thoracic aorta. Western blot method was used to detect the ABCA1, ABCG1, LXRa, CD36, PPARy, NFKB expression in the thoracic aorta. Immunohistochemical method was used to detect the ABCA1, LXRa, CD36, PPARy expression in the liver.Results:1. Blood lipid levelComparing to the normal control group, mice in the model control group had significant higher level of TG, LDL, VLDL (p=0.003,0.001,0.004), and significant lower level of ApoA-Ⅰ, HDL (p=0.001,0.007) in the serum; no significant differences were found between the two groups on the level of TC and ox-LDL; comparing to the model control group, mice in the moxibustion group had significant lower TG and LDL (p=0.03,0.001), significant higher ApoA-1(p=0.001), while no significant differences were found between the two groups on VLDL and HDL (p=0.11,0.11); comparing to the model control group, mice in the moxa smoke group had significant lower TG and LDL (p=0.01,0.008), significant higher ApoA-Ⅰ (p=0.01), no significant difference were found on VLDL, HDL (p=0.24,0.11); no significant differences were found between the moxibustion group and the moxa smoke group (p>0.05) 2. Inflammatory cytokines and NFKBComparing to the normal control group, mice in the model control group had significant higher level of IL-1, IL-6, TNFa, hs-CRP, vWF, ICAM-1, VCAM-1, MCP-1(p<0.001), significant lower level of IL-10(p<0.001) in the serum; comparing to the model control group, mice in the moxibustion group had significant lower IL-1, IL-6, TNFa, hs-CRP, vWF, ICAM-1, VCAM-1, MCP-1(p=0.005,0.001,0.001,0.01,0.001,0.001,0.001,0.002), significant higher level of IL-10(p<0.001); comparing to the model control group, mice in the moxa smoke group had significant lower IL-1, IL-6, TNFa, hs-CRP, vWF, ICAM-1, VCAM-1, MCP-1(p=0.003,0.003,0.001,0.01,0.02,0.03,0.001,0.005), significant higher level of IL-10(p=0.01); no significant differences were found between the moxibustion group and the moxa smoke group (p>0.05).There were no significant differences among all groups on NFKB expression in the aorta.3. Plaque area ratio in the aortaThe plague area ratio in the aorta root were:15.76%in the model control group,8.55%in the moxa smoke group,7.34%in the moxibustion group. The plague area ratio in the thoracic aorta were:9.05%in the model control group,4.37%in the moxa smoke group,2.47%in the moxibustion group.Comparing to the model control group, mice in the moxibustion group had significant lower plague area ratio both in the aorta root and the thoracic aorta (p=0.04,0.01); the difference between the moxa smoke group and model control group in the plague area ratio was not significant(p=0.08,0.08); No significant differences were found between the moxibustion group and the moxa smoke group (p>0.05)4. Expression of CD36, ABCA1, ABCG1, LXRa, PPARy in the aortaComparing to the normal control group, mice in the model control group had significant lower ABCA1、ABCG1. LXRa expression(p<0.001,0.04,0.004), no significant different CD36、PPARy expression in the thoracic aorta(p>0.05); comparing to the model control group, mice in the moxibustion group had significant higher ABCA1、LXRa expression (p=0.05,0.03), no significant different ABCG1experssion(p=0.14), comparing to the model control group, mice in the moxa smoke group had significant higher ABCA1, LXRa expression (p<0.01,0.02), no significant different ABCG1experssion(p=0.14); no significant differences were found in the ABCA1, ABCG1, LXRa expression between the moxibustion group and the moxa smoke group (p>0.05).5. Expression of CD36, ABCA1, LXRa, PPARy in the liverComparing to the normal control group, mice in the model control group had significant lower ABCA1, PPARγ expression(P<0.001,<0.001), significant higher CD36, LXRa expression (p=0.004,0.002); ABCA1, PPARy expression were significant higher (p=0.03,0.03) while the LXRa expression was significant lower (p=0.03) in the moxibusiton group than in the model conrol group, the CD36expression showed no significant difference between the two groups(p=0.09); ABCA1, PPARy expression were significant higher (p=0.002,0.01) while the CD36, LXRa expression was significant lower (p=0.02,0.02) in the moxibusiton group than in the model conrol group; no significant differences were found in the ABCA1, CD36, LXRa, PPARy expression between the moxibustion group and moxa smoke group (p>0.05).Conclusions:1. Moxibustion could alleviate atherosclerosis, inhibit the growth of plague, and moxa somke also have this tendency.2. Moxibustion and moxa smoke could regulate lipid metabolism and promote the cholesterol efflux at the lesion area and improve the metabolic function of liver, which may be one of the mechanisms of their anti-atherosclerosis effect.3. Moxibustion and moxa smoke could inhibite inflammation in the body, which may be one of the mechanisms of their anti-atherosclerosis effect.更多还原