Effects of <italic style="font-style: italic">Lizhong Tongmai </italic>acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice

SUN Rui; CHENG Zedong; LI Di; YIN Jingyao

doi:10.13703/j.0255-2930.20230606-0001

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Effects of Lizhong Tongmai acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice

Study on Mechanism | 更新时间：2024-06-03

- Effects of Lizhong Tongmai acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice
- Chinese Acupuncture & Moxibustion Vol. 44, Issue 2, Pages: 169-174(2024)
- 作者机构：
  
  辽宁中医药大学针灸推拿学院，沈阳 110847
- 作者简介：
- 基金信息：
- DOI：10.13703/j.0255-2930.20230606-0001
  CLC：
- Received：06 June 2023，
  
  Online First：15 November 2023，
  
  Published：12 February 2024
- 稿件说明：
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孙瑞,成泽东,李迪等.“理中通脉”法针刺对动脉粥样硬化小鼠TMAO、CD36表达及胆固醇沉积的影响[J].中国针灸,2024,44(02):169-174.

SUN Rui,CHENG Zedong,LI Di,et al.Effects of Lizhong Tongmai acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice[J].Chinese Acupuncture & Moxibustion,2024,44(02):169-174.
孙瑞,成泽东,李迪等.“理中通脉”法针刺对动脉粥样硬化小鼠TMAO、CD36表达及胆固醇沉积的影响[J].中国针灸,2024,44(02):169-174. DOI： 10.13703/j.0255-2930.20230606-0001.

SUN Rui,CHENG Zedong,LI Di,et al.Effects of Lizhong Tongmai acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice[J].Chinese Acupuncture & Moxibustion,2024,44(02):169-174. DOI： 10.13703/j.0255-2930.20230606-0001.

摘要

目的

观察“理中通脉”法针刺对动脉粥样硬化（AS）小鼠氧化三甲胺（TMAO）、CD36表达及胆固醇沉积的影响，探讨电针治疗AS的可能机制。

方法

SPF级C57BL/6J 雄性ApoE-/-小鼠31只以高脂饲料喂养8周制备AS模型，造模成功后将剩余30只小鼠随机分为模型组、西药组和电针组，每组10只，另有同品系普通小鼠10只作为空白组。空白组和模型组不予干预；西药组采用阿托伐他汀钙片稀释液灌胃给药；电针组取“内关”“天枢”“足三里”进行电针干预，同侧“内关”与“足三里”、“天枢”与小鼠尾巴连接电针仪，选择疏密波，频率2 Hz /15 Hz，电流强度0.3 mA，留针10 min，每次针单侧，左右交替，隔日１次。均干预11周。干预结束后，HE 染色法观察小鼠主动脉病理形态；微板法检测小鼠三酰甘油（TG）、总胆固醇（TC）、低密度脂蛋白胆固醇（LDL-C）、高密度脂蛋白胆固醇（HDL-C）含量；超高效液相色谱-质谱联用技术法检测小鼠血浆TMAO含量；Western blot法检测小鼠主动脉CD36蛋白表达水平；微量法检测小鼠主动脉胆固醇脂（CE）含量，计算CE/TC比值。

结果

与空白组比较，模型组小鼠出现明显的动脉粥样硬化病理性改变，血清TG、TC、LDL-C含量升高（

0.01），HDL-C含量降低（

0.01），血浆TMAO含量、主动脉CE含量及CE/TC比值升高（

0.01），主动脉CD36蛋白表达水平升高（

0.01）；与模型组比较，西药组和电针组小鼠动脉病理改变有所改善，血清TG、TC、LDL-C含量降低（

0.05），HDL-C含量升高（

0.05），血浆TMAO含量、主动脉CE含量及CE/TC比值降低（

0.01），CD36蛋白表达水平降低（

0.05）；电针组血清TG、TC含量高于西药组（

0.05）。

结论

“理中通脉”法针刺可改善AS小鼠动脉病理性改变及血脂水平，降低血浆TMAO含量，抑制主动脉CD36蛋白表达，减少胆固醇沉积，这可能是电针治疗AS的作用机制之一。

Abstract

Objective

To observe the effects of

Lizhong Tongmai

acupuncture （acupuncture for regulating middle

jiao

and promoting meridians） on trimethylamine-N-oxide (TMAO)

CD36 expression

and cholesterol deposition in atherosclerotic (AS) mice

exploring potential mechanism of electroacupuncture (EA) in treating AS.

Methods

A total of 31 male SPF-grade C57BL/6J ApoE-/- mice were fed with high-fat diet for 8 weeks to establish AS model. After successful modeling

the remaining 30 mice were randomly divided into a model group

a medication group

and an EA group

with 10 mice in each group. An additional 10 normal mice of the same strain were selected as a blank group. The mice in the blank group and the model group received no intervention. The mice in the medication group were treated with intragastric administration of atorvastatin calcium. The mice in the EA group were treated with EA at "Neiguan" (PC 6)

"Tianshu" (ST 25) and "Zusanli" (ST 36). The same-side "Neiguan" (PC 6) and "Zusanli" (ST 36)

"Tianshu" (ST 25) and the tail of the mice were connected to the EA apparatus

with disperse-dense wave

a frequency of 2 Hz/15 Hz

and a current intensity of 0.3 mA for 10 min per session. Acupuncture was performed unilaterally per session

alternating between the left and right sides

with a frequency of once every other day. After intervention

HE staining was used to observe the pathological morphology of the aorta. Microplate assays were conducted to measure triglyceride (TG)

total cholesterol (TC)

low-density lipoprotein cholesterol (LDL-C)

and high-density lipoprotein cholesterol (HDL-C) levels in serum. Ultra high performance liquid chromatography-mass spectrometry technique （UPLC-MS） was employed to detect TMAO level in plasma. Western blot was performed to assess CD36 protein expression level in the aorta. Microanalysis was used to measure cholesterol ester (CE) level in the aorta and the CE/TC ratio was calculated.

Results

Compared with the blank group

the mice in the model group exhibited significant pathological changes of atherosclerosis

serum TG

LDL-C levels were increased (

0.01)

and HDL-C level was decreased (

0.01); the plasma TMAO level

aortic CE level

and the CE/TC ratio were increased (

0.01)

along with elevated CD36 protein expression level in the aorta (

0.01). Compared with the model group

the mice in the medication group and the EA group showed improvements in aortic pathology

serum TG

LDL-C levels were reduced

HDL-C levels were increased (

0.05); plasma TMAO levels

aortic CE levels

and the CE/TC ratio were decreased (

0.01)

and CD36 protein expression levels were lowered (

0.05). The serum TG and TC levels in the EA group were higher than those in the medication group (

0.05).

Conclusion

The

Lizhong Tongmai

acupuncture can ameliorate aortic pathological changes

regulate blood lipid levels

reduce plasma TMAO level

inhibit CD36 protein expression in the aorta

and decrease cholesterol deposition. These effects may contribute to the therapeutic mechanism of EA in treating AS.

关键词

Keywords

references

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⁰

Effects of Lizhong Tongmai acupuncture on TMAO, CD36 expression, and cholesterol deposition in atherosclerotic mice

DOI：10.13703/j.0255-2930.20230606-0001

摘要

Abstract

关键词

Keywords

references