不同频次温和灸对动脉粥样硬化兔血脂及TNF-α、MCP-1、ICAM-1含量的影响

Effects of Different Frequency Mild Moxibustion on Blood Lipid and Contents of TNF-α, MCP-1 and ICAM-1 in Rabbits with Atherosclerosis

目的 观察不同频次温和灸对动脉粥样硬化(AS)家兔血脂及肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)、细胞间黏附分子-1(ICAM-1)含量的影响,探讨不同频次温和灸治疗AS的可能机制。方法 将50只家兔适应性喂养1周后,随机分为空白组13只和造模组37只。空白组给予标准饲料饲养,造模组给予高脂饲料饲养8周并结合免疫损伤的方法制备AS模型。造模结束后每组随机选取1只家兔进行血脂检测和主动脉弓部组织病理形态结构观察,检验造模是否成功。将造模成功后的家兔随机分为模型组、低频次艾灸组、高频次艾灸组,每组12只。从第9周开始,低频次艾灸组和高频次艾灸组选取“膻中”“神阙”“内关(双)”“血海(双)”穴位进行艾灸干预,低频次艾灸组治疗隔日1次,高频次艾灸组治疗每日1次,每次治疗30 min,共干预4周。空白组、模型组仅进行抓取、固定,不予干预。干预结束后,观察家兔一般情况;酶法测定血清甘油三酯(TG)、总胆固醇(TC)含量;比色法测定血清低密度脂蛋白胆固醇(LDL-C)含量;苏丹Ⅳ染色观察各组家兔主动脉弓部组织病理形态结构,电镜观察主动脉超微结构;酶联免疫法检测血清TNF-α、MCP-1、ICAM-1含量。结果 与空白组比较,模型组家兔活动、食欲、皮毛、大便形态积分均增高(P<0.01);血清TC、TG、LDL-C含量均增高(P<0.01),主动脉弓部组织结构界限不清,脂质浸润明显,平滑肌形态不完整;血清TNF-α、MCP-1、ICAM-1含量均升高(P<0.001)。与模型组比较,低频次艾灸组家兔活动、食欲、皮毛积分均降低(P<0.05),大便形态积分显著降低(P<0.01),血清TC、TG、LDL-C含量均降低(P<0.01),主动弓部组织结构较清晰,有少量炎性细胞浸润,内膜较光滑,平滑肌细胞排列欠整齐;血清TNF-α、MCP-1、ICAM-1含量均降低(P<0.01)。高频次艾灸组家兔活动、食欲、皮毛、大便形态积分均降低(P<0.05);血清TC、TG、LDL-C含量均降低(P<0.01),主动脉弓部组织结构界限清晰,未见明显炎性细胞浸润,内膜较光滑,平滑肌细胞排列稍欠整齐;血清TNF-α、MCP-1含量均降低(P<0.01),ICAM-1含量显著降低(P<0.001)。与低频次艾灸组比较,高频次艾灸组活动、皮毛积分均降低(P<0.05),食欲积分低于低频次艾灸组(P<0.01);血清LDL-C降低(P<0.01),TG、TC含量均降低,但差异无统计学意义(P>0.05);血清TNF-α、ICAM-1含量均降低(P<0.05),MCP-1含量显著降低(P<0.01)。结论 高频次温和灸更能有效调节AS家兔血脂水平,改善主动脉形态和超微结构,可能与艾灸能降低相关炎性因子TNF-α、MCP-1和ICAM-1的含量有关。

Objective To observe the effects of different frequency mild moxibustion on blood lipid and contents of TNF-α,monocyte chemoattractant protein 1(MCP-1) and intercellular adhesion molecule-1(ICAM-1) in rabbits with atherosclerosis(AS)andexplore the possible mechanism of different frequency mild moxibustion in the treatment of AS.Methods After adaptive feeding for one week, 50 rabbits were randomly divided into blank group(n=13) and model group(n=37).The blank group was fed with standard diet, and the model group was fed with high-fat diet for 8 weeks, and the AS model was prepared with the method of immune injury.At the end of modeling, one rabbit in each group was randomly selected for blood lipid detection and histopathological morphology and structure observation of aortic arch to test whether the modeling was successful.The rabbits were randomly divided into model group, low frequency moxibustion group and high frequency moxibustion group, with 12 rabbits in each group.Starting from the 9th week, low-frequency moxibustion group and high-frequency moxibustion group selected “Danzhong(RN17)”,“Shenque(RN8)”,“Neigan(PC6)(double)” and “Xuehai(SP10)(double)” for moxibustion intervention, low-frequency moxibustion group was treated once every other day, high-frequency moxibustion group was treated once a day, each treatment was 30min, a total of 4 weeks of intervention.The blank group and model group were only grabbed and fixed without intervention.After the intervention, the general situation of rabbits was observed.The contents of serum triglyceride(TG) and total cholesterol(TC) were determined by enzymatic method.Serum low density lipoprotein cholesterol(LDL-C) was determined by colorimetry.The pathological morphology of aortic arch was observed by Sudan Ⅳ staining, and the ultrastructure of aorta was observed by electron microscopy.Serum levels of TNF-α,MCP-1 and ICAM-1 were detected by ELISA.Results Compared with blank group, the activity, appetite, fur and stool morphology scores in the model group were increased(P<0.01).The serum levels of TC,TG and LDL-C were increased(P<0.01),aortic arch tissue structure boundary was not clear, lipid invasion was obvious, smooth muscle morphology was incomplete, and there were migration cells.The levels of TNF-α,MCP-1 and ICAM-1 in serum were increased(P<0.001).Compared with those of the model group, the activity, appetite and fur score in low-frequency moxibustion group were decreased(P<0.05),stool shape score was significantly decreased(P<0.01),serum levels of TC,TG and LDL-C were decreased(P<0.01),active arch tissue structure was clearer, a small number of inflammatory cells infiltrated, and the intima was smooth.The contents of TNF-α,MCP-1 and ICAM-1 in serum decreased when smooth muscle cells were not arranged neatly(P<0.01).In the high-frequency moxibustion group, the activity, appetite, fur and stool morphology scores were decreased(P<0.05),serum levels of TC,TG and LDL-C were decreased(P<0.01),the active arch tissue structure boundary was clear, no obvious inflammatory cell infiltration was observed, the intima was smooth, smooth muscle cells arranged slightly less orderly.The levels of TNF-α and MCP-1 in serum were decreased(P<0.01),and the content of ICAM-1 was significantly decreased(P<0.001).Compared with those of the low-frequency moxibustion group, the activity and fur score in the high-frequency moxibustion group were decreased(P<0.05),the appetite score was lower than that in the low-frequency moxibustion group(P<0.01),serum LDL-C was decreased(P<0.01),the levels of TG and TC were decreased, but the differences were not statistically significant(P>0.05).The contents of TNF-α and ICAM-1 in serum were decreased(P<0.05),and the content of MCP-1 was significantly decreased(P<0.01).Conclusion High frequency mild moxibustion can more effectively regulate the level of blood lipid and improve the morphology and ultrastructure of aorta in AS rabbits, which may be related to that moxibustion can reduce the contents of related inflammatory factors TNF-α,MCP-1 and ICAM-1.