陳曉龍，孫 陽(yáng)，楊新林

(1. 廣西工業(yè)職業(yè)技術(shù)學(xué)院基礎(chǔ)教學(xué)部，廣西南寧 530001；2. 重慶醫(yī)科大學(xué)體育工作部，重慶 400010；3. 西北農(nóng)林科技大學(xué)體育部，陜西楊凌 712100)

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蘋(píng)果原花青素對(duì)小鼠運(yùn)動(dòng)能力和抗氧化能力的影響

陳曉龍1，孫 陽(yáng)2*，楊新林3

(1. 廣西工業(yè)職業(yè)技術(shù)學(xué)院基礎(chǔ)教學(xué)部，廣西南寧 530001；2. 重慶醫(yī)科大學(xué)體育工作部，重慶 400010；3. 西北農(nóng)林科技大學(xué)體育部，陜西楊凌 712100)

為研究蘋(píng)果中原花青素對(duì)小鼠運(yùn)動(dòng)能力和抗氧化能力的影響，選取5周齡健康雄性小鼠45只，隨機(jī)分為3組，進(jìn)行3周適應(yīng)性游泳訓(xùn)練。小鼠運(yùn)動(dòng)結(jié)束后，通過(guò)輕度麻醉?xiàng)l件下采集血液和肌組織,檢測(cè)血清中乳酸(LD)和丙二醛(MDA)含量，以及測(cè)定超氧化物歧化酶(SOD)和過(guò)氧化氫酶(CAT)的活力。結(jié)果表明,各組小鼠的初始體重對(duì)比無(wú)顯著差異(P>0.05);運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠的游泳至力竭的時(shí)間明顯長(zhǎng)于安靜對(duì)照組；與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的游泳至力竭的時(shí)間也呈現(xiàn)明顯的延長(zhǎng)趨勢(shì)(P<0.05)。游泳運(yùn)動(dòng)以后，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠血清和肌組織中LD和MDA含量明顯低于安靜對(duì)照組；與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的血清LD和MDA含量也呈現(xiàn)明顯下降趨勢(shì)(P<0.05)。另外，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠SOD和CAT活性明顯高于安靜對(duì)照組，且與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組SOD和CAT活性顯著升高(P<0.05)。說(shuō)明蘋(píng)果原花青素能提高小鼠運(yùn)動(dòng)能力和抗氧化能力，且與有氧運(yùn)動(dòng)相結(jié)合能夠產(chǎn)生協(xié)同效應(yīng)，具有較好的應(yīng)用前景。

蘋(píng)果原花青素；運(yùn)動(dòng)能力；抗氧化能力；小鼠

原花青素(Proanthocyanidins)作為一種天然抗氧化劑，其來(lái)源相當(dāng)廣泛，常見(jiàn)的葡萄、銀杏、紫甘藍(lán)等果實(shí)的中原花青素含量極高[1-4]。原花青素對(duì)機(jī)體作用顯著，具有明顯抗氧化、抗癌等多種療效[5]。既往研究已經(jīng)證實(shí)原花青素在機(jī)體功能調(diào)節(jié)上的相關(guān)作用，有研究顯示原花青素的應(yīng)用相比其他抗氧化劑作用更為顯著，能改善一些由自由基引起的損傷，如動(dòng)脈粥樣硬化、冠心病、糖尿病、腫瘤，以及抗衰老等[6-10]。針對(duì)自由基，已有研究表明，大強(qiáng)度急性運(yùn)動(dòng)會(huì)造成大量的自由基的產(chǎn)生，若機(jī)體清除能力有限，體內(nèi)殘留的多余自由基很容易發(fā)生氧化反應(yīng)，造成肌肉損傷和運(yùn)動(dòng)疲勞；與此同時(shí)，自由基還可引起血紅蛋白等合成障礙，進(jìn)一步影響運(yùn)動(dòng)能力[11-12]。蘋(píng)果營(yíng)養(yǎng)價(jià)值高，富含多種營(yíng)養(yǎng)物質(zhì)，原花青素是蘋(píng)果中主要酚類(lèi)化合物[13]。鑒于原花青素對(duì)機(jī)體的無(wú)損害性、無(wú)不良反應(yīng)，安全性極高[14]。本文旨在研究蘋(píng)果原花青素與有氧運(yùn)動(dòng)對(duì)小鼠運(yùn)動(dòng)能力和抗氧化能力的影響，為原花青素的推廣使用提供科學(xué)依據(jù)。

1 材料與方法

1.1 材料

1.1.1 實(shí)驗(yàn)動(dòng)物 5周齡健康雄性昆明小鼠,體重15 g～25 g，購(gòu)于廣西醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)室,共45只，進(jìn)行分籠適應(yīng)性喂養(yǎng)，提供正常飲食。動(dòng)物實(shí)驗(yàn)方法和操作程序符合實(shí)驗(yàn)動(dòng)物管理與使用指南(國(guó)家生命科學(xué)委員會(huì)，美國(guó)國(guó)立衛(wèi)生研究院出版6-23，2012修訂)。

1.1.2 蘋(píng)果原花青素制備 蘋(píng)果原花青素樣品由廣西醫(yī)科大學(xué)動(dòng)物實(shí)驗(yàn)室提供。采用冷凍干燥儀器進(jìn)行冷凍干燥，持續(xù)12 h，獲得原花青素。提純后低溫保存，置于-20℃低溫箱中保存。

1.2 方法

1.2.1 動(dòng)物分組及訓(xùn)練 小鼠在20℃～26℃環(huán)境下飼養(yǎng)，相對(duì)濕度為40%～50%。適應(yīng)性喂養(yǎng)持續(xù)1周，隨后進(jìn)行為期3天的無(wú)負(fù)重的適應(yīng)性游泳訓(xùn)練。將小鼠隨機(jī)分為3組，每組15只小鼠。安靜對(duì)照組：不進(jìn)行水池的游泳訓(xùn)練，在游泳前2 h給予同等劑量的生理鹽水灌服；運(yùn)動(dòng)對(duì)照組：進(jìn)行水池游泳訓(xùn)練，在游泳前2 h給予同等劑量的生理鹽水灌服；試驗(yàn)組：進(jìn)行水池游泳訓(xùn)練，在游泳前2 h給予原花青素0.01 mg/g體重的灌胃補(bǔ)充。除安靜對(duì)照組，其他兩組均進(jìn)行游泳訓(xùn)練，訓(xùn)練模型參考等人的方法操作，并在實(shí)際環(huán)境中根據(jù)具體情況進(jìn)行適當(dāng)調(diào)節(jié)。游泳訓(xùn)練持續(xù)3周(泳缸水溫28℃～30℃，水深約為35 cm)，第1周為無(wú)負(fù)重游泳(3 min/d)，第2周和第3周小鼠每天游泳時(shí)間逐周遞增10 min，并進(jìn)行負(fù)重游泳(小鼠尾部懸掛等同體重的5%的鉛絲)。安靜對(duì)照組保持正常生理活動(dòng)。

1.2.2 小鼠樣本制備 小鼠運(yùn)動(dòng)結(jié)束后，即可通過(guò)輕度麻醉?xiàng)l件下進(jìn)行血液采集。小鼠血清采用眼眶采血方式，血量為1.5 mL作用。隨后在4℃環(huán)境下離心20 min(3 000 r/min)，采集上清液，低溫保存。同時(shí)，小鼠輕度麻醉后進(jìn)行斷髓處死方式，取小鼠骨骼肌組織，用生理鹽水洗凈，制備組織勻漿，采用與血清標(biāo)本相同的離心模式。樣本備置以供下用。

1.2.3 血清和肌組織LD、MDA、SOD和CAT含量的檢測(cè) 按照上述步驟進(jìn)行3 000 r/min離心5 min后，取出低溫保存的分離血清和肌組織。通過(guò)使用紫外可見(jiàn)分光官渡及測(cè)量吸光度，檢測(cè)血清中乳酸(LD)和丙二醛(MDA)含量，以及測(cè)定超氧化物歧化酶(SOD)和過(guò)氧化氫酶(CAT)的活力。

1.2.4 統(tǒng)計(jì)學(xué)方法 采用SPSS 22.0 統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)處理，計(jì)量資料采用均值 ± 標(biāo)準(zhǔn)差的形式表示，各組間比較采用方差分析。P<0.05 為差異有統(tǒng)計(jì)學(xué)意義。

2 結(jié)果

2.1 3周藥物補(bǔ)給及運(yùn)動(dòng)訓(xùn)練后小鼠體重變化

如表1所示，各組小鼠的初始體重對(duì)比，無(wú)顯著差異(P>0.05)，且各組小鼠在試驗(yàn)期間(為期3周)

體重呈現(xiàn)穩(wěn)定增長(zhǎng)趨勢(shì)，但并未發(fā)現(xiàn)顯著(P>0.05)。游泳訓(xùn)練運(yùn)動(dòng)開(kāi)始后，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組的小鼠體重明顯低于安靜對(duì)照組，但差異不顯著(P<0.05)。同時(shí)，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組的小鼠體重?zé)o顯著差異(P<0.05)。

2.2 原花青素及運(yùn)動(dòng)訓(xùn)練對(duì)小鼠運(yùn)動(dòng)至力竭時(shí)間的影響

方差分析統(tǒng)計(jì)分析顯示，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠的游泳至力竭的時(shí)間明顯長(zhǎng)于安靜對(duì)照組(88.34±6.34 ,114.62±8.38 vs 69.56±4.57)，差異顯著(P<0.05)；同時(shí)，與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的游泳至力竭的時(shí)間也呈現(xiàn)明顯的延長(zhǎng)趨勢(shì)(114.62±8.38 vs 88.34±6.34)，差異顯著(P<0.05)。

2.3 各組小鼠血清和肌組織中LD，MDA，SOD和CAT含量的變化

游泳運(yùn)動(dòng)以后，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠血清中LD和MDA含量明顯低于安靜對(duì)照組；同時(shí)，與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的血清LD和MDA含量也呈現(xiàn)明顯下降趨勢(shì)(P<0.05，表2)。另外，從表2可以看出，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠SOD和CAT活性明顯高于安靜對(duì)照組，且與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組SOD和CAT活性顯著升高(P<0.05)。表3顯示，肌組織中LD/MDA/SOD和CAT變化趨勢(shì)與血清檢測(cè)的各指標(biāo)趨勢(shì)相似(P<0.05)。

表1 各組小鼠體重變化及組間對(duì)比情況

注：*與安靜對(duì)照組相比，P<0.05；**與運(yùn)動(dòng)對(duì)照組相比，P<0.05。

Note: * Compared with the sedentary group,P<0.05; ** Compared with the exercise group,P<0.05.

表2 各組小鼠血清LD、MDA、SOD和CAT變化及組間對(duì)比情況

注：*與安靜對(duì)照組相比，P<0.05；**與運(yùn)動(dòng)對(duì)照組相比，P<0.05。

Note: * Compared with the sedentary group,P<0.05; ** Compared with the exercise group,P<0.05.

表3 各組小鼠肌組織LD/MDA/SOD和CAT變化及組間對(duì)比情況

注：*與安靜對(duì)照組相比，P<0.05；**與運(yùn)動(dòng)對(duì)照組相比，P<0.05。

Note: * Compared with the sedentary group,P<0.05; ** Compared with the exercise group,P<0.05.

3 討論

本文旨在研究蘋(píng)果原花青素在小鼠運(yùn)動(dòng)能力和抗氧化能力方面的作用。研究結(jié)果顯示各組小鼠的初始體重對(duì)比無(wú)明顯差異，而在游泳訓(xùn)練運(yùn)動(dòng)開(kāi)始后，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組的小鼠體重明顯低于安靜對(duì)照組，說(shuō)明游泳訓(xùn)練有效抑制了小鼠體重的上升。同時(shí)，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組的小鼠體重?zé)o明顯差異，進(jìn)一步證實(shí)蘋(píng)果原花青素對(duì)小鼠的體重增長(zhǎng)無(wú)明顯影響。本文關(guān)于原花青素及運(yùn)動(dòng)訓(xùn)練對(duì)小鼠運(yùn)動(dòng)至力竭時(shí)間的影響結(jié)果顯示，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠的游泳至力竭的時(shí)間明顯長(zhǎng)于安靜對(duì)照組，表明經(jīng)過(guò)3周的游泳運(yùn)動(dòng)，小鼠運(yùn)動(dòng)能力顯著增強(qiáng)；同時(shí)，與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的游泳至力竭的時(shí)間也呈現(xiàn)明顯的延長(zhǎng)趨勢(shì)，提示補(bǔ)充蘋(píng)果原花青素與適量運(yùn)動(dòng)更能增強(qiáng)小鼠的運(yùn)動(dòng)能力。蘋(píng)果原花青素與游泳訓(xùn)練對(duì)增強(qiáng)小鼠運(yùn)動(dòng)能力具有協(xié)同效應(yīng)，結(jié)合更能顯著增強(qiáng)小鼠運(yùn)動(dòng)能力。該結(jié)果的機(jī)制可能是適量游泳訓(xùn)練提高了蘋(píng)果原花青素的吸收，最大程度發(fā)揮原花青素的抗氧化能力。以往研究已經(jīng)證實(shí)，適量運(yùn)動(dòng)對(duì)于增強(qiáng)機(jī)體的氧化應(yīng)激，系統(tǒng)血氧供應(yīng)等作用均明顯[16]，相比于劇烈運(yùn)動(dòng)，更能夠避免自由基和乳酸的大量產(chǎn)生，對(duì)于增強(qiáng)機(jī)體運(yùn)動(dòng)能力更有顯著效果[17]，同時(shí)綜合應(yīng)用蘋(píng)果原花青素，也能明顯提高機(jī)體抗氧化能力，減輕小鼠機(jī)體的過(guò)氧化損傷，提示原花青素在機(jī)體抗氧化能力方面作用顯著。

本文研究結(jié)果進(jìn)一步顯示，游泳運(yùn)動(dòng)以后，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠血清和肌組織中LD和MDA含量明顯低于安靜對(duì)照組；同時(shí)，于運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組小鼠的血清LD和MDA含量也呈現(xiàn)明顯下降趨勢(shì)。此外，運(yùn)動(dòng)對(duì)照組和試驗(yàn)組小鼠SOD和CAT活性明顯高于安靜對(duì)照組，且與運(yùn)動(dòng)對(duì)照組相比，試驗(yàn)組SOD和CAT活性顯著升高。眾所周知，SOD和CAT是主要的酶類(lèi)抗氧化劑[18]，有氧運(yùn)動(dòng)和蘋(píng)果原花青素的聯(lián)合使用促進(jìn)SOD和CAT等抗氧化酶的升高；同時(shí)，其他指標(biāo)如LD和MDA含量的變化也與機(jī)體自由基的產(chǎn)生以及機(jī)體抗氧化能力密切相關(guān)[19]。金雯等[20]研究表明,有氧運(yùn)動(dòng)可提高骨骼肌SOD活性，從而提高機(jī)體抗氧化能力以及抗自由基作用。另外，LD含量的變化與運(yùn)動(dòng)相關(guān)，而相比于無(wú)氧運(yùn)動(dòng)，有氧運(yùn)動(dòng)能夠避免乳酸的產(chǎn)生，LD的含量相較更低。試驗(yàn)組血清和肌組織LD和MDA含量均呈下降趨勢(shì)，而SOD和CAT活性均呈現(xiàn)上升趨勢(shì)，進(jìn)一步揭示蘋(píng)果原花青素在抗氧化方面的功能。綜上所述，蘋(píng)果原花青素均能提高小鼠運(yùn)動(dòng)能力和抗氧化能力，且與有氧運(yùn)動(dòng)相結(jié)合能夠產(chǎn)生協(xié)同效應(yīng)，具有較好的應(yīng)用前景。

[1] Bai Y F, Liu S, Liu Y M. The antioxidant activity and synergistic effects of lycopene, grape seed oil, V E and procyanidine[J]. Food Sci Technol, 2014，39(9)：222-227.

[2] Feng L L, Liu B X, Zhong J Y, et al. Effect of grape procyanidins on tumor angiogenesis in liver cancer xenograft models[J]. Asian Pacific J Cancer Prevention, 2014, 15(2):737-741.

[3] Huang H B, Liu C H, Jin Z M, et al.Preparation and antioxidant activity of proanthocyanidins fromGinkgobilobaleaves[J]. Chin J Mod Appl Pharm, 2016, 33(6):686-690.

[4] Li S L, Chen X M, Lin J. Optimization of microwave-assisted extracting of proanthocyanidins from purple cabbage and evaluation of antioxidant activity in vitro[J]. Adv Materials Res, 2012, 490(4):3500-3504.

[5] Gültekin-zgüven M, jlal Berkta,zelik B. Influence of processing conditions on procyanidin profiles and antioxidant capacity of chocolates: Optimization of dark chocolate manufacturing by response surface methodology[J]. LWT - Food Sci Technol, 2016, 66:252-259.

[6] Yang H, Xiao L, Yuan Y, et al. Procyanidin B2 inhibits NLRP3 inflammasome activation in human vascular endothelial cells.[J]. Biochem Pharmacol, 2014, 92(4):599-606.

[7] Yi M. A major invention to change the fate of hundreds of millions of cardiovascular and cerebrovascular patients original health pillow proanthocyanidins[J]. Modern Marketing: Business Edition, 2015(4). doi: 10.2147/VHRM.S59429.

[8] Yin W, Li B, Li X, et al. Anti-inflammatory effects of grape seed procyanidin B2 on a diabetic pancreas.[J]. Food & Function, 2015, 6(9):3065-3071.

[9] Mao J T, Xue B, Smoake J, et al. MicroRNA-19a/b mediates grape seed procyanidin extract-induced anti-neoplastic effects against lung cancer.[J]. J Nutri Biochem, 2016, 34:118-125.

[10] Yang Q S, Liu L, Wang Z. Research Progress on antioxidant ability of proanthocyanidins to aging organism[J]. Modern Agri Sci Technol, 2009 (21):290-298.

[11] Tan D X, Hardeland R, Manchester L C, et al. Cyclic-3-hydroxymelatonin (C3HOM), a potent antioxidant, scavenges free radicals and suppresses oxidative reactions.[J]. Cur Med Chem, 2014, 21(13):1557-65.

[12] Chang C Q, Jin S S, Lv Y R. Effect of nutrition on free radical injury induced by exercise[J]. Chinese J Rehabil Med, 2014, 29(6):580-584.

[13] Fu C, Yang D, Peh W Y, et al. Structure and antioxidant activities of proanthocyanidins from elephant apple (dillenia indica linn.).[J]. J Food Sci, 2015, 80(10):C2191-C2199.

[14] Zhang H W, Zhang Y, Ma C M. Research progress of proanthocyanidins[J]. Food Sci, 2015, 36(5):296-304.

[15] Arimitsu T, Matsuura R, Yunoki T, et al. Relationship between oxygen uptake and oxygen supply system during constant-load supine exercise[J]. Biol Sport, 2010, 28(2):151-156.

[16] Mairb X H. Red blood cells in sports: effects of exercise and training on oxygen supply by red blood cells[J]. Front Physiol, 2013, 4(3):332.

[17] Wang R X. Effects of free radicals on human body motor ability[J]. Contemp Sports Sci Technol, 2014, 4(22):22-23.

[18] Meng S L, Chen J Z, Xu P, et al. Hepatic antioxidant enzymes SOD and CAT of Nile tilapia (Oreochromisniloticus) in response to pesticide methomyl and recovery pattern.[J]. Bul Environ Contam Toxicol, 2014, 92(4):388-392.

[19] Trivedi D S, Nandlal D, Mahdi D A A, et al. Evaluation of antioxidant enzymes activity and malondialdehyde levels in chronic periodontitis patients with diabetes.[J]. J Periodontol, 2014, 85(5):713-720.

[20] Jin W, Li M X, Qian Hai, et al. Effects of aerobic exercise on antioxidant capacity of skeletal muscle in aging rats[J]. China Gerontol, 2011, 31(14):2706-2707.

Effects of Proanthocyanidins from Apples on Exercise Ability and Antioxidant Capacity in Mice

CHEN Xiao-long1，SUN Yang2，YANG Xin-lin3
(1.BasicTeachingDepartmentofGuangxiInstituteofIndustryTechnology，Nanning，Guangxi， 530001，China；2.SportsDepartmentofMedicalUniversityofChongqing,Chongqing,400010，China；3.SportsDepartment,NorthwestAgricultureandForestryUniversity,Yangling,Shaanxi,712100，China)

To study the effects of proanthocyanidins from apples on exercise ability and antioxidant capacity in mice, a total of 45 healthy male mice aged 5 weeks were randomly divided into three groups, and trained with adaptive swimming for 3 weeks in each group. After the exercise of mice, serum and muscular tissues were collected under mild anesthesia. Subsequently, serum lactic acid (LD) and malondialdehyde (MDA) contents, and superoxide dismutase (SOD) and catalase (CAT) activities were then determined. Results showed that there was no significant difference in the initial weight of each group (allP>0.05). The exercise control group and experimental group mice exhaustive swimming time were significantly longer than the sedentary control group; at the same time, compared with the exercise control group, the exhaustive swimming time in the mice of the the experimental group also showed obvious prolonged trend (P<0.05). In addition, LD and MDA of serum and muscle tissues were significantly lower in the exercise control group and experimental group than sedentary control group; compared with the exercise control group, serum LD and MDA contents in mice of experimental group were also decreased (P<0.05). In addition, the activities of SOD and CAT in the exercise control group and the experimental group were significantly higher than those in the sedentary control group, and the activities of SOD and CAT in the experimental group were significantly higher than those in the exercise control group (allP<0.05). In conclusion,apple proanthocyanidins can improve the exercise ability and antioxidant capacity of mice, and combined with aerobic exercise can produce synergistic effect, having good application prospects.

apple proanthocyanidins; exercise ability; antioxidant capacity；mouse

2016-12-23

陳曉龍(1983―)，男，浙江衢州人，碩士，講師，主要從事運(yùn)動(dòng)醫(yī)學(xué)、體育教學(xué)、運(yùn)動(dòng)訓(xùn)練。 *通訊作者

S852.2

A

1007-5038(2017)06-0057-04