崔瑩雪,趙百孝,劉鈞天,黃玉海

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艾灸及艾煙對(duì)ApoE-/-小鼠血脂、肝臟病理形態(tài)及肝臟CD36、ABCA1表達(dá)的影響

崔瑩雪1,趙百孝2,劉鈞天2,黃玉海2

(1.首都醫(yī)科大學(xué)附屬北京中醫(yī)醫(yī)院,北京 100010;2.北京中醫(yī)藥大學(xué)針灸推拿學(xué)院,北京 100029)

目的 觀察艾灸及艾煙干預(yù)對(duì)動(dòng)脈粥樣硬化模型血脂、肝臟病理改變和肝細(xì)胞內(nèi)膽固醇代謝相關(guān)分子的影響,分析艾灸和艾煙對(duì)膽固醇代謝的調(diào)節(jié)作用,探討艾灸及艾煙的作用機(jī)制。方法 將51只8周齡ApoE-/-小鼠隨機(jī)分為模型組、艾煙組、艾灸組,每組17只,20只C57BL/6小鼠作為空白對(duì)照。正常組、模型組小鼠常規(guī)抓取和固定;艾煙組小鼠暴露于10 mg/m3～15 mg/m3艾煙環(huán)境;艾灸組小鼠艾灸關(guān)元穴。所有干預(yù)每日20 min,每星期6次,連續(xù)干預(yù)12星期。全自動(dòng)生化分析儀檢測(cè)總膽固醇);HE染色觀察肝臟病理形態(tài);免疫組化法檢測(cè)肝臟CD36、ABCA1的表達(dá)。結(jié)果 模型組小鼠血清TG、LDL-C含量顯著高于正常組(=0.003,=0.001),HDL-C含量顯著低于正常組(=0.007),TC含量與正常組無顯著差異(＞0.05);艾灸組小鼠血清TG、LDL-C含量顯著低于模型組(=0.03,=0.001),HDL-C、TC含量與模型組無顯著差異(=0.11,=0.11);艾煙組小鼠血清TG、LDL-C含量顯著低于模型組(=0.01,=0.008),HDL-C含量與模型組無顯著差異(＞0.05);艾灸組與艾煙組各項(xiàng)血脂指標(biāo)均無顯著性差異(＞0.05)。模型組小鼠肝細(xì)胞索、肝竇排列紊亂,肝細(xì)胞腫脹變性明顯。艾煙組、艾灸組小鼠肝細(xì)胞腫脹程度明顯減輕,炎細(xì)胞浸潤(rùn)程度較模型組減輕。模型組CD36表達(dá)顯著高于正常組(=0.004),ABCA1表達(dá)顯著低于正常組(＜0.001);艾灸組CD36表達(dá)與模型組無顯著性差異(=0.09),ABCA1表達(dá)顯著高于模型組(=0.03);艾煙組CD36表達(dá)顯著低于模型組(=0.02),ABCA1表達(dá)顯著高于模型組(=0.002);艾灸組與艾煙組CD36、ABCA1表達(dá)均無顯著差異(＞0.05)。結(jié)論 早期艾灸關(guān)元穴可以一定程度調(diào)整動(dòng)脈粥樣硬化模型ApoE-/-小鼠血脂代謝紊亂的狀態(tài),減緩肝臟病變發(fā)生,減少肝臟內(nèi)膽固醇的蓄積,這可能是艾灸療法預(yù)防動(dòng)脈粥樣硬化的作用機(jī)制之一。艾煙作為艾灸生成物,是艾灸發(fā)揮治療作用的有效因素。

艾條灸;艾煙;ApoE-/-小鼠;膽固醇

心腦血管疾病是人類目前發(fā)病率最高的疾病之一,具有極高的致死率和致殘率。動(dòng)脈粥樣硬化(atherosclerosis,AS)是心血管疾病的主要病理基礎(chǔ),其病變的發(fā)生發(fā)展是一個(gè)復(fù)雜的過程,膽固醇代謝紊亂是導(dǎo)致AS的主要發(fā)病機(jī)制。肝臟是脂質(zhì)代謝的重要器官,在脂類的消化、吸收、合成、分解與運(yùn)輸中都發(fā)揮重要作用。體內(nèi)脂質(zhì)過多會(huì)增加肝臟負(fù)荷,脂類物質(zhì)在肝細(xì)胞內(nèi)蓄積,引起肝臟代謝障礙,肝臟功能受損會(huì)進(jìn)一步導(dǎo)致脂質(zhì)代謝紊亂,加重形成動(dòng)脈粥樣硬化性病變。

中醫(yī)學(xué)認(rèn)為動(dòng)脈粥樣硬化由痰濁血瘀互結(jié),脈絡(luò)痹阻不通引起[1-3],屬于中醫(yī)艾灸療法的適應(yīng)證范圍[4-6]。根據(jù)中醫(yī)學(xué)理論,艾灸有溫通百脈、宣通氣血的功能。大量的臨床和實(shí)驗(yàn)研究發(fā)現(xiàn),艾灸療法具有降低血脂、改善動(dòng)脈粥樣硬化的作用[7-9]。艾煙是艾灸過程中的產(chǎn)物,其中含有多種活性成分,具有抗炎、抗氧化等作用。研究發(fā)現(xiàn),艾煙可以良性調(diào)整機(jī)體膽固醇代謝[10-11]。

本實(shí)驗(yàn)選用AS動(dòng)物模型ApoE-/-小鼠(載脂蛋白E基因敲除)作為觀察對(duì)象,并對(duì)其分別進(jìn)行艾灸關(guān)元穴和艾煙暴露兩種干預(yù)。通過觀察ApoE-/-小鼠血脂、肝臟的病理形態(tài),以及肝細(xì)胞內(nèi)CD36和ABCA1(肝細(xì)胞內(nèi)參與膽固醇代謝的重要分子)的表達(dá),分析艾灸和艾煙對(duì)膽固醇代謝的調(diào)節(jié)作用,探討艾灸及艾煙的作用機(jī)制。

1 材料與方法

1.1 實(shí)驗(yàn)動(dòng)物

本實(shí)驗(yàn)選用8周齡ApoE-/-小鼠作為AS模型,同齡相同遺傳背景非轉(zhuǎn)基因小鼠C57BL/6作為空白對(duì)照。小鼠購(gòu)自于北京大學(xué)醫(yī)學(xué)部實(shí)驗(yàn)動(dòng)物科學(xué)部。ApoE-/-小鼠采用高脂飼料(北京維通利華實(shí)驗(yàn)動(dòng)物有限公司提供,含15%豬油、2%膽固醇、0.05%膽酸)喂養(yǎng),C57BL/6采用常規(guī)飼料喂養(yǎng)。所有小鼠均可自由進(jìn)食飲水,飼養(yǎng)環(huán)境溫度為(22±2)℃,濕度為50%～60%。采用人工控制室內(nèi)照明,保持12 h光照(8:00- 20:00)和黑暗(20:00-次日8:00)交替循環(huán)。

1.2 主要儀器與試劑

細(xì)艾條(0.5 cm×20 cm),購(gòu)自河南南陽漢醫(yī)艾絨有限公司;小鼠固定器(150 mm×65 mm×50 mm),可固定小鼠頭、尾、四肢部位,且底板處設(shè)有小孔,可暴露腹部,購(gòu)自上海華巖儀器設(shè)備有限公司;帶孔玻璃缸,自制,80 cm×80 cm×60 cm,頂層玻璃正中設(shè)有直徑為5 mm圓孔;光散射式數(shù)字粉塵測(cè)試儀,購(gòu)自北京賓達(dá)綠創(chuàng)科技有限公司;羅氏MODULAR P800全自動(dòng)生化分析儀;總膽固醇(TC)、甘油三酯(TG)、高密度脂蛋白膽固醇(HDL-C)、低密度脂蛋白膽固醇(LDL-C)試劑盒購(gòu)自北京北化康泰臨床試劑有限公司;ABCA1(ab18180)及CD36(ab325031)一抗均購(gòu)自ABCAM公司。

1.3 動(dòng)物分組及處理

51只8周齡ApoE-/-小鼠隨機(jī)分為模型組、艾煙組、艾灸組,每組17只,20只C57BL/6小鼠作為空白對(duì)照組(正常組)。小鼠適應(yīng)性喂養(yǎng)1星期后,進(jìn)行實(shí)驗(yàn)干預(yù),干預(yù)方法如下。

正常組采用固定器固定小鼠頭、尾、四肢部位,不進(jìn)行其他干預(yù);模型組采用固定器固定小鼠頭、尾、四肢部位,不進(jìn)行其他干預(yù);艾煙組將小鼠放入固定器后放入玻璃缸,玻璃缸內(nèi)臵有調(diào)試好的粉塵測(cè)試儀。點(diǎn)燃艾條,從玻璃缸上方圓孔中將燃燒的一端插入,令艾煙充滿玻璃缸。用粉塵測(cè)試儀測(cè)試其濃度,根據(jù)課題組前期測(cè)定臨床艾灸診室的平均濃度[12],以及既往實(shí)驗(yàn)報(bào)道的艾煙有效濃度[13],本實(shí)驗(yàn)中艾煙濃度的設(shè)定在10～15 mg/m3,當(dāng)艾煙濃度達(dá)到預(yù)期濃度范圍時(shí)將艾條取出,迅速封閉圓孔。此后每隔3 min記錄一次濃度數(shù)值,使數(shù)值保持在10～15 mg/m3之間。若數(shù)值高于15 mg/m3,可將上方玻璃蓋稍微移動(dòng),令艾煙散去部分后迅速將蓋子移回;若數(shù)值低于10 mg/m3,再將點(diǎn)燃的艾條由圓孔插入以補(bǔ)充艾煙。

艾灸組采用固定器固定小鼠頭、尾、四肢部位后架起。固定器底板處設(shè)有小孔,可暴露腹部關(guān)元穴,點(diǎn)燃艾條放于小鼠腹部下方,使灸火透過固定器底部小孔傳至小鼠關(guān)元穴。同時(shí)在施灸過程中對(duì)實(shí)驗(yàn)操作間的艾煙濃度進(jìn)行監(jiān)測(cè),艾煙濃度保持在10～15 mg/m3之間。

所有干預(yù)均為每日20 min,每星期6次,連續(xù)干預(yù)12星期,在第12星期末處死小鼠。

1.4 取材及檢測(cè)

末次干預(yù)后,禁食不禁水12 h。小鼠麻醉后,用眼球摘除法從眼眶靜脈叢采集血液標(biāo)本約1.5～2 mL,置于離心管中,靜置1 h后,離心15 min(4℃,1500 r/min)。離心后將血清保存于﹣20℃冰箱中備用。采用羅氏(MODULAR P800)全自動(dòng)生化分析儀檢測(cè)總膽固醇(TC)、甘油三酯(TG)和高密度脂蛋白膽固醇(HDL-C)、低密度脂蛋白膽固醇(LDL-C)。

迅速解剖腹部,取下肝臟,生理鹽水沖洗,放入4%中性多聚甲醛固定24 h,制作石蠟切片,并進(jìn)行HE染色和免疫組化反應(yīng)。免疫組化二步法具體步驟,將切片脫蠟至水;采用枸櫞酸緩沖液放入高壓鍋水煮進(jìn)行抗原修復(fù);雙氧水封閉;滴加一抗(抗體稀釋度均為1:100),0.01 mol的PBS代替一抗做陰性對(duì)照,4℃冰箱過夜;滴加二抗(即用型)30 min;室溫DAB顯色,顯微鏡下觀察,觀察目標(biāo)出現(xiàn)陽性后,蒸餾水洗終止反應(yīng);蘇木素復(fù)染,鹽酸乙醇分化,氨水返藍(lán);脫水透明及中性樹膠封片。

染色完畢后,在Olympus BX51光學(xué)顯微鏡下觀察,采用Anymicro DSSTM圖像采集系統(tǒng)讀片并拍照。每組隨機(jī)選取10只小鼠,將每只小鼠切片高倍鏡下拍攝三個(gè)視野,將采集的圖像應(yīng)用Image-pro Plus5.1圖像分析系統(tǒng)進(jìn)行分析。所得數(shù)據(jù)作免疫組織化學(xué)染色陽性信號(hào)累積光密度(IOD),以IOD值作為組織表達(dá)的定量標(biāo)準(zhǔn)。累積光密度(IOD)=平均光密度(AOD)×陽性面積(A)。

1.5 統(tǒng)計(jì)學(xué)方法

采用SPSS13.0統(tǒng)計(jì)軟件對(duì)實(shí)驗(yàn)結(jié)果進(jìn)行統(tǒng)計(jì)分析。各組數(shù)據(jù)均符合方差分析條件,采用單因素方差分析(-),繼以檢驗(yàn)法進(jìn)行組間比較,結(jié)果以均數(shù)±標(biāo)準(zhǔn)差表示。所有檢驗(yàn)均為雙側(cè)檢驗(yàn),若＜0.05認(rèn)為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié)果

2.1 艾灸及艾煙對(duì)ApoE-/-小鼠血脂的影響

各組小鼠均有個(gè)別死亡現(xiàn)象,對(duì)各組存活小鼠血脂水平比較結(jié)果顯示(見表1),模型組小鼠血清TG、LDL-C含量顯著高于正常組(=0.003,=0.001), HDL-C含量顯著低于正常組(=0.007),TC含量與正常組無顯著差異(＞0.05);艾灸組小鼠血清TG、LDL-C含量顯著低于模型組(=0.03,=0.001),HDL-C、TC含量與模型組無顯著差異(=0.11,=0.11);艾煙組小鼠血清TG、LDL含量顯著低于模型組(=0.01,=0.008), HDL含量與模型組無顯著差異(＞0.05);艾灸組與艾煙組各項(xiàng)血脂指標(biāo)均無顯著性差異(＞0.05)。

表1 各組小鼠血清TC、TG、LDL-C、HDL-C含量比較 (±,mmol/L)

組別nTCTGLDL-CHDL-C 正常組191.38±0.310.23±0.070.35±0.120.61±0.12 模型組121.63±0.240.50±0.251)0.57±0.162)0.45±0.121) 艾煙組151.44±0.230.26±0.143)0.39±0.113)0.55±0.12 艾灸組161.47±0.210.29±0.163)0.33±0.074)0.55±0.11

注:與正常組比較1)＜0.05,2)＜0.001;與模型組比較3)＜0.05,4)＜0.001

2.2 艾灸及艾煙對(duì)ApoE-/-小鼠肝臟病理形態(tài)的影響

正常組小鼠肝細(xì)胞索、肝竇、中央靜脈、匯管區(qū)均細(xì)胞形態(tài)如常,排列有序;未見細(xì)胞變性、腫脹,匯管區(qū)未見膽管增生、炎細(xì)胞浸潤(rùn)。模型組小鼠肝細(xì)胞索、肝竇排列紊亂,肝細(xì)胞腫脹變性明顯,肝竇受壓變窄,中央靜脈迂曲擴(kuò)張充血,匯管區(qū)血管擴(kuò)張明顯,炎細(xì)胞浸潤(rùn)。艾煙組小鼠肝細(xì)胞索排列較規(guī)則,肝細(xì)胞輕度腫脹,匯管區(qū)血管充血,炎細(xì)胞浸潤(rùn)。艾灸組小鼠肝細(xì)胞索排列較規(guī)則,肝細(xì)胞腫脹程度明顯減輕,匯管區(qū)血管充血,少量炎細(xì)胞浸潤(rùn)(見圖1)。

圖1 各組小鼠肝臟病理形態(tài)(×20)

2.3 艾灸及艾煙對(duì)各組ApoE-/-小鼠肝臟CD36、ABCA1表達(dá)的影響

與正常組小鼠比較,模型組小鼠肝臟CD36表達(dá)顯著高于正常組(=0.004),ABCA1表達(dá)顯著低于正常組(＜0.001)。與模型組小鼠比較,艾灸組小鼠肝臟CD36表達(dá)與模型組無顯著性差異(=0.09),ABCA1表達(dá)顯著高于模型組(=0.03)。艾煙組小鼠肝臟CD36表達(dá)顯著低于模型組(=0.02),ABCA1表達(dá)顯著高于模型組(=0.002);艾灸組與艾煙組CD36、ABCA1表達(dá)均無顯著差異(=0.32,=0.15)。詳見表2,圖2-3。

表2 各組小鼠肝臟CD36、ABCA1累積光密度值(IOD)比較 (±)

組別nCD36ABCA1 正常組101574.27±268.611817.00±276.60 模型組101784.63±317.401)1344.13±241.822) 艾煙組101607.87±360.343)1539.57±269.963) 艾灸組101661.03±245.571477.70±248.293)

注:與正常組比較1)＜0.05,2)＜0.001;與模型組比較3)＜0.05

圖2 各組小鼠肝臟CD36表達(dá)比較(×20)

圖3 各組小鼠肝臟ABCA1表達(dá)比較(×20)

4 討論

肝臟是脂質(zhì)代謝的重要器官,飲食攝取的脂質(zhì),經(jīng)由肝臟合成極低密度膽固醇(VLDL)和甘油三酯,并轉(zhuǎn)運(yùn)入血,進(jìn)一步水解供機(jī)體利用,同時(shí)體內(nèi)過多的膽固醇,可以在高密度脂蛋白(HDL)的運(yùn)載下重新回到肝臟(此過程稱為膽固醇的逆向運(yùn)轉(zhuǎn)),在肝臟水解,并以膽汁的形式排泄。血液中過高的膽固醇水平,不僅會(huì)引發(fā)動(dòng)脈粥樣硬化病變,同時(shí)會(huì)對(duì)肝臟組織造成巨大的負(fù)擔(dān)。體內(nèi)過多的脂質(zhì)可引起肝臟細(xì)胞脂質(zhì)過度攝入及合成,同時(shí)對(duì)脂質(zhì)的運(yùn)轉(zhuǎn)能力下降,脂類物質(zhì)在肝細(xì)胞內(nèi)蓄積,引起肝臟代謝障礙,造成肝臟組織病理?yè)p傷,肝臟的實(shí)質(zhì)細(xì)胞受累成為脂肪負(fù)荷細(xì)胞,成為脂肪肝。并可引發(fā)一系列脂質(zhì)代謝紊亂,加重動(dòng)脈粥樣硬化病變?？梢哉f,動(dòng)脈粥樣硬化和肝臟功能障礙密切相關(guān)。有實(shí)驗(yàn)研究表明,高脂飲食可以導(dǎo)致家兔在主動(dòng)脈和肝臟都出現(xiàn)病理改變。并隨著高脂飲食的時(shí)間延長(zhǎng),病變逐漸加重[14]。本實(shí)驗(yàn)發(fā)現(xiàn),ApoE-/-小鼠在給予高脂飲食12星期的情況下,出現(xiàn)肝細(xì)胞索、肝竇排列紊亂,肝細(xì)胞明顯腫脹變性,匯管區(qū)血管擴(kuò)張,炎性細(xì)胞浸潤(rùn)等現(xiàn)象,可以證明小鼠存在病變趨勢(shì)。相較于模型組,早期進(jìn)行艾灸或艾煙干預(yù)的ApoE-/-小鼠,肝細(xì)胞的腫脹變性及炎性細(xì)胞浸潤(rùn)等現(xiàn)象較未治療的小鼠減輕,說明艾灸治療或艾煙暴露存在保護(hù)肝細(xì)胞、防止肝細(xì)胞變性的作用趨勢(shì)。但本實(shí)驗(yàn)各組小鼠肝臟均未出現(xiàn)明顯的脂肪變性、壞死的情況出現(xiàn),對(duì)肝臟的病理形態(tài)的影響有待于更長(zhǎng)時(shí)間的實(shí)驗(yàn)觀察。

B族清道夫受體CD36廣泛表達(dá)于多種細(xì)胞類型,介導(dǎo)細(xì)胞對(duì)膽固醇的攝取。在巨噬細(xì)胞中,CD36可介導(dǎo)巨噬細(xì)胞對(duì)氧化低密度脂蛋白的攝取,參與巨噬細(xì)胞泡沫化。肝細(xì)胞表面的CD36可負(fù)責(zé)肝臟對(duì)脂肪酸的攝取,CD36在肝細(xì)胞的基礎(chǔ)表達(dá)較低,但在高脂飲食的刺激下其表達(dá)可顯著增加[15]。CD36的過表達(dá)可增加組織對(duì)脂肪酸和膽固醇攝入、儲(chǔ)存和利用。Koonen DP等[15]研究發(fā)現(xiàn),高脂喂養(yǎng)可使小鼠肝CD36蛋白表達(dá)增加2.6倍,肝臟甘油三酯存儲(chǔ)上升1.7倍。有研究顯示,高脂飲食引起肝臟CD36的表達(dá)增加是脂肪肝病變的重要潛在病理機(jī)制[15-16]。臨床觀察也發(fā)現(xiàn),肝臟CD36基因表達(dá)與脂肪肝患者的肝脂肪含量呈正相關(guān)[17]。在本實(shí)驗(yàn)中發(fā)現(xiàn),ApoE-/-小鼠肝臟CD36的表達(dá)較正常小鼠明顯升高,而經(jīng)艾煙干預(yù)的ApoE-/-小鼠肝臟CD36明顯降低,提示艾煙可以防止肝細(xì)胞對(duì)脂質(zhì)的過度攝取,減輕肝臟脂肪性病變。艾灸治療組的ApoE-/-小鼠,肝臟CD36的表達(dá)有降低趨勢(shì),但是效果不像艾煙干預(yù)一樣明顯。對(duì)于艾煙效果優(yōu)于艾灸治療的現(xiàn)象,還有待于進(jìn)一步的研究進(jìn)行深入探討。

ABCA1屬于ATP結(jié)合盒(ABC)轉(zhuǎn)運(yùn)蛋白A類亞族,主要在細(xì)胞質(zhì)膜處表達(dá)[18],可介導(dǎo)膽固醇穿過細(xì)胞膜,促進(jìn)細(xì)胞內(nèi)游離膽固醇外流[19-22]。ABCA1廣泛表達(dá)于多種不同組織中,尤其在肝、腸及巨噬細(xì)胞中表達(dá)較高[23-27]。肝臟ABCA1可以促進(jìn)細(xì)胞內(nèi)膽固醇流向載脂蛋白A-I (ApoA-I),并促進(jìn)新生ApoA-I的酯化、形成HDL[28]。肝臟是HDL的主要合成場(chǎng)所,研究顯示,肝臟ABCA1對(duì)調(diào)節(jié)血液中HDL-C水平具有重要作用,研究顯示特異性敲除小鼠肝臟ABCA1的表達(dá),可以顯著降低約80%血漿HDL的水平,造成體內(nèi)膽固醇逆向轉(zhuǎn)運(yùn)功能下降[29-30]。在本實(shí)驗(yàn)研究中發(fā)現(xiàn),ApoE-/-小鼠肝臟ABCA1表達(dá)下降,提示肝細(xì)胞內(nèi)存在膽固醇流出障礙,從而造成小鼠體內(nèi)膽固醇蓄積和動(dòng)脈粥樣硬化病變。艾灸和艾煙干預(yù)可以顯著提高小鼠肝臟ABCA1的表達(dá),說明二者具有促進(jìn)膽固醇逆向運(yùn)轉(zhuǎn)、減輕肝臟膽固醇蓄積的作用趨勢(shì)。艾煙和艾灸兩種干預(yù)方法在提高肝臟ABCA1表達(dá)的效果上沒有體現(xiàn)明顯差異。

本研究結(jié)果表明,早期艾灸關(guān)元穴在一定程度可以調(diào)整ApoE-/-小鼠血脂代謝紊亂的狀態(tài),減緩小鼠肝臟病變發(fā)生,并有抑制肝臟的CD36表達(dá),促進(jìn)ABCA1表達(dá)的作用趨勢(shì)。減少肝臟內(nèi)膽固醇的蓄積,可能是艾灸療法預(yù)防動(dòng)脈粥樣硬化的作用機(jī)制之一。艾煙作為艾灸生成物,是艾灸發(fā)揮治療作用的有效因素。

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Effects of Moxibustion and Moxa Smoke on Blood Lipids, and Hepatic Pathologic Morphology and CD36 and ABCA1 Expressions in ApoE-/-mice

1,2,2,2.

1.100010,; 2.100029,

Objective To investigate the intervention effects of moxibustion and moxa smoke on blood lipids, hepatic pathological changes and intrahepatocytic molecules related to cholesterol metabolism and analyze the regulating effects of moxibustion and moxa smoke on cholesterol metabolism and explore the mechanisms of actions of moxibustion and moxa smoke. Method Fifty-one 8-week-old ApoE-/-mice were randomized into model, moxa smoke and moxibustion groups, 17 mice each. Twenty C57BL/6 mice comprised a blank control group. The normal and model groups of mice were routinely grabbed and fastened. The moxa smoke group of mice was exposed to 10-15 mg/m3moxa smoke circumstances. The moxibustion group of mice was given moxibustion on point Guanyuan(CV4). All interventions were made 20 min daily, 6 times a week, for 12 consecutive weeks. Total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured using an automatic biochemical analyzer. Hepatic pathologic morphology was observed by HE staining. Hepatic CD36 and ABCA1 expressions were determined by immunohistochemical method. Result In the model group of mice, serum TG and LDL-C contents were significantly higher than in the normal group (=0.003;=0.001); HDL-C content was significantly lower than in the normal group (=0.007); TC content had no significant difference compared with the normal group (＞0.05). In the moxibustion group of mice, serum TG and LDL-C contents were significantly lower than in the model group (=0.03;=0.001) and HDL-C content had no significant difference compared with the model group (=0.11). In the moxa smoke group of mice, serum TG and LDL contents were significantly lower than in the model group (=0.01;=0.008) and HDL content had no significant difference compared with the model group (=0.11). There were no significant differences in various blood lipid indicators between the moxibustion and moxa smoke groups (＞0.05). There were hepatic cell cord and sinusoid derangement and obvious hepatocytic swelling in the model group of mice. In the moxa smoke and moxibustion groups, hepatocytic swelling subsided significantly, and inflammatory cell infiltration reduced compared with the model group. In the model group,CD36 expression was significantly higher than in the normal group (=0.004) and ABCA1 expression was significantly lower than in the normal group (=0.001). In the moxibustion group, CD36 expression had no significant difference compared with the model group (=0.09) and ABCA1 expression was significantly higher than in the model group (=0.03). In the moxa smoke group, CD36 expression was significantly lower than in the normal group (=0.02) and ABCA1 expression was significantly higher than in the model group (=0.002). There were no significant differences in CD36 and ABCA1 expressions between the moxibustion and moxa smoke groups (＞0.05). Conclusion Early moxibustion on point Guanyuan can regulate disorders of blood lipid metabolism, delay the occurrence of hepatic lesions and reduce intrahepatic accumulation of cholesterol to a certain extent in an ApoE-/-mouse model of atherosclerosis. That may be one of the mechanisms by which moxibustion therapy prevents atherosclerosis. Moxa smoke as the product of moxibustion is an effective factor in moxibustion producing a therapeutic effect.

Moxa stick moxibustion; Moxa smoke; ApoE-/-mice; Cholesterol

R2-03

A

10.13460/j.issn.1005-0957.2016.08.1008

2015-12-20

國(guó)家自然科學(xué)基金面上項(xiàng)目(81373730)

崔瑩雪(1986 - ),女,住院醫(yī)師,博士

趙百孝(1963 - ),男,教授,Email:baixiao100@vip.sina.com