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Effects of Curcumin on Sperm Motility, Viability, Mitochondrial Activity and Plasma Membrane Integrity in Boar Semen
Biomed Sci Letters 2017;23:406-410
Published online December 31, 2017;  https://doi.org/10.15616/BSL.2017.23.4.406
© 2017 The Korean Society For Biomedical Laboratory Sciences.

A-Sung Lee1, Sang-Hee Lee2, Seunghyung Lee1, and Boo-Keun Yang1,†

1College of Animal Life Sciences, Chuncheon 24341, Korea,
2Institute of Animal Resources, Kangwon National University, Chuncheon 24341, Korea
Correspondence to: Boo-Keun Yang. College of Animal Life Science, Kangwon National University, Chuncheon 24341, Korea. Tel: +82-33-250-8623, Fax: +82-33-259-5572, e-mail: bkyang@kangwon.ac.kr
Received September 5, 2017; Revised October 13, 2017; Accepted October 13, 2017.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
 Abstract

Curcumin is known as a natural antioxidant, decreasing oxidative stress in animal cells. Generally, oxidative stress induces reactive oxygen species in sperm and leads to decreased sperm characteristics in pigs. Therefore, this study investigated the influence of curcumin on sperm motility, viability, mitochondrial activity and plasma membrane integrity in pigs. Curcumin (0, 5 and 10 μM) was treated in boar semen, which were incubated for 9 hours in 37°C. Then, motility, viability, mitochondrial activity, plasma membrane integrity of sperm was analyzed every 3 hours. In the results, sperm motility was significantly increased by 5 μM curcumin after 3 and 9 hours after incubation, and viability was significantly higher in 5 μM curcumin treatment at 3 hours (P0.05). Similarly, sperm mitochondrial activity and plasma membrane integrity were significantly increased by 5 μM curcumin at 3, 6 and 9 hours after incubation (P0.05). There results suggest that curcumin improve sperm characteristics such as motility, viability, mitochondrial activity, and plasma membrane integrity, and may exert a positive effect on sperm fertility in pigs.

Keywords : Curcumin, Motility, Viability, Plasma membrane integrity, Sperm
Body

넂 닔以쓽 솢꽦궛냼뒗 DNA 넀긽, 吏吏 怨쇱궛솕 諛 떒諛깆쭏 蹂꽦쓣 룷븿븳 꽭룷 궡 留롮 븙쁺뼢쓣 겮移섎ʼn 궛솕 뒪듃젅뒪瑜 쑀諛쒖떆耳 궓꽦쓽 遺덉엫쓣 쑀諛쒗븯뒗 二쇱슂 썝씤쑝濡 븣젮졇 엳떎(Ko et al., 2014). 씠윭븳 솢꽦궛냼瑜 젣嫄고빐二쇰뒗 빆궛솕젣뒗 룷쑀룞臾쇱뿉 엳뼱 닔而룹쓽 깮떇 湲곌怨 꽭룷瑜 蹂댄샇븯뒗 뿭븷쓣 븯湲 븣臾몄뿉 踰덉떇 뒫젰쓣 뼢긽떆궎湲 쐞븳 빆궛솕젣쓽 뿰援щ뒗 솢諛쒗븯寃 吏꾪뻾릺怨 엳떎(Ciftci et al., 2011; Głombik et al., 2014).

Curcumin (1,7-bis(4-hydroxy-3-methoxy phenyl)-1,6-heptadiene-3,5-dione) Curry굹 Turmeric (Curcuma longa)뿉 룷븿릺뒗 솴깋 븞猷 꽦遺꾩쑝濡, 깮臾쇳븰쟻 듅꽦뿉 븳 뿰援ш 떎뼇븳 諛⑸㈃뿉꽌 씠猷⑥뼱吏怨 엳떎(Tayyem et al., 2006; He et al., 2015; Vallianou et al., 2015). 듅엳 curcumin 룷쑀룞臾쇱뿉 엳뼱 빆궛솕, 빆뿼利, 빆떦눊 諛 빆븫 옉슜쓣 븯뒗 以묒슂븳 臾쇱쭏濡 븣젮졇 엳떎(Aggarwal et al., 2007; El-Bahr 2013; Ghosh et al., 2015). 떎젣濡 curcumin cadmium 諛 imidacloprid뿉 끂異쒕맂 伊먯뿉꽌 젙옄瑜 蹂댄샇(Lonare et al., 2015; Oguzturk, 2012)븯硫, 2,3,7,8-Tetracholorodibenzo-p-dioxin(TCDD)뿉 쓽븳 궛솕쟻 넀긽쓣 媛먯냼(Ciftci et al., 2011)떆궓떎怨 븣젮졇 엳떎. 삉븳 curcumin di-2-ethylhexyl phthalate뿉 쓽븳 꽭룷 궗硫몄쓣 셿솕떆耳 젙옄 슫룞꽦뿉 湲띿젙쟻씤 쁺뼢(Głombik et al., 2014)쓣 以 肉먮쭔 븘땲씪 닔而룹쓽 깮떇꽭룷瑜 蹂댄샇븯뒗 뿭븷(Gobbo et al., 2015)븳떎怨 븣젮졇 엳湲 븣臾몄뿉 curcumin씠 궛솕 뒪듃젅뒪濡쒕꽣 쑀諛쒕릺뒗 踰덉떇 湲곕뒫 諛 遺덉엫 벑뿉 湲띿젙쟻씤 슚怨쇰 誘몄튌 寃껋쑝濡 蹂댁뿬吏꾨떎. 븯吏留 뤌吏 젙옄뿉꽌 curcumin쓣 궗슜븯뒗뜲 엳뼱 쟻빀븳 냽룄 쟻젅븳 끂異쒖떆媛꾩뿉 븳 뿰援щ뒗 蹂닿퀬릺吏 븡븯湲 븣臾몄뿉 蹂 뿰援ъ뿉꽌뒗 뤌吏 젙옄瑜 씠슜븯뿬 泥쒖뿰 빆궛솕젣씤 curcumin쓽 슚怨쇰 寃利앺븯怨 理쒖쟻쓽 냽룄 諛 끂異쒖떆媛꾩뿉 븯뿬 븣븘蹂닿퀬옄 뿰援щ 떎떆븯떎.

紐⑤뱺 룞臾쇱떎뿕 媛뺤썝븰援 룞臾쇱떎뿕쑄由ъ쐞썝쉶쓽 듅씤(KIACUC-09-0139)쓣 뼸뼱 닔뻾븯怨, 蹂 떎뿕뿉꽌 궗슜맂 curcumin (C1386) Sigma-Aldrich Chemical Company(St. Louis, MO, USA)濡쒕꽣 援ъ엯븯뿬 궗슜븯떎. 떆猷뚯쓽 닔吏묒쓣 쐞븯뿬 Duroc, Yorkshire 諛 Landrace媛 援먯옟맂 닔而 뤌吏뿉꽌 쓬寃쎌닔븬踰뺤쓣 씠슜빐 젙븸쓣 梨꾩랬븯쑝硫, 뤌吏 젙븸씗꽍븸씤 modena (Lee et al., 2017)濡 샎빀븯뿬 씗꽍 젙븸쓽 냽룄瑜 1.0 × 107媛/mL媛 릺룄濡 븳 썑 17°C쓽 긽깭濡 2떆媛 씠궡 떎뿕떎濡 슫諛섑븯뿬 떎뿕뿉 궗슜븯떎. 씠 썑 37°C, 5% CO2 議곌굔뿉꽌 珥 9떆媛 룞븞 젙옄瑜 諛곗뼇븯硫 3떆媛 媛꾧꺽쑝濡 젙옄쓽 슫룞꽦, 깮議댁쑉, 誘명넗肄섎뱶由ъ븘 솢꽦 諛 썝삎吏덈쭑쓽 삩쟾꽦 遺꾩꽍쓣 떎떆븯떎. 젙옄쓽 슫룞꽦쓣 愿李고븯湲 쐞븯뿬 37°C쓽 媛삩뙋뿉꽌 10遺 씠긽 媛삩떆궓 slide glass뿉 젙븸 떆猷 20 μL瑜 뼥뼱쑉由 썑 cover glass瑜 뜮 뮘, 쐞긽李 쁽誘멸꼍(Nicon, Japan)쑝濡 200 諛곗쑉 븯뿉 슫룞꽦쓣 寃궗븯쑝硫, 젙옄쓽 誘몃媛 吏곸씠뒗 寃껋쓣 슫룞씠 엳뒗 젙옄濡 뙋떒븯떎.

젙옄쓽 깮議댁쑉 寃궗뒗 Hoechest 33342 (HO) / propidium iodide (PI) 뿼깋 諛⑸쾿쓣 궗슜븯떎. 100 μL 젙븸 떆猷뚯뿉 0.5 mg/mL PI 슜븸쓣 10 μL 泥④ 썑 37°C뿉꽌 5遺꾧컙 젙移섏떆궓 썑, 0.5 mg/mL HO 슜븸쓣 10 μL瑜 泥④ 썑 37°C뿉꽌 10遺꾧컙 젙移섏떆耳곕떎. 씠 썑 뿼깋맂 젙븸 떆猷 20 μL瑜 slide glass뿉 遺꾩< 썑 epifluorescence microscope (Ziess, Germany)瑜 씠슜빐 400 諛곗쑉 460/500 nm 뙆옣 븯뿉꽌 愿李고븯떎. 궡븘엳뒗 젙옄뒗 뙆깋, 二쎌 젙옄뒗 遺됱깋쑝濡 뙋떒븯 떎.

誘명넗肄섎뱶由ъ븘 湲곕뒫 寃궗瑜 쐞븯뿬 Rhodamin123 (R123) / PI 뿼깋 諛⑸쾿쓣 씠슜(Lee and Park, 2015)븯쑝硫, 1 mL쓽 젙옄 떆猷뚯뿉 3 μL쓽 0.5 mg/mL R123 슜븸쓣 泥④븳 뮘 37°C뿉꽌 15遺꾧컙 젙移섏떆궓 썑 0.5 mg/mL PI 슜븸쓣 10 μL 꽔怨 10遺꾧컙 異붽쟻쑝濡 諛곗뼇븯떎. 誘명넗肄섎뱶由ъ븘 湲곕뒫 epifluorescence microscope (Ziess)濡 400 諛곗쑉 븯뿉 R123 490/515 nm 뙆옣뿉꽌 PI뒗 545/590 nm뿉꽌 愿李고븯떎. 以묓렪遺媛 끃깋 삎愿묒쓣 쓣뒗 젙옄媛 誘명넗肄섎뱶由ъ븘쓽 젙긽 湲곕뒫쓣 븯뒗 寃껋쑝濡 뙋떒븯떎.

젙옄쓽 썝삎吏덈쭑 湲곕뒫 寃궗뒗 hypoosmotic swelling test(HOST)濡 寃궗븯怨, 젙븸 50 μL 1 mL쓽 hypo-osmotic solution (624.9 mM Na-citrate 諛 74.9 mM fructose)쓣 샎빀 썑 37°C뿉꽌 30遺꾧컙 諛곗뼇븯떎. 씠 썑 slide glass뿉 떆猷 20 μL瑜 뼥뼱쑉由 썑 cover glass瑜 뜮 뮘, 쐞긽李 쁽誘멸꼍(Nicon)쑝濡 200 諛곗쑉 븯뿉꽌 寃궗븯쑝硫, 젙옄쓽 瑗щ━媛 媛먭꺼 엳嫄곕굹, 굹꽑삎쑝濡 媛먭꺼졇 엳쑝硫 썝삎吏덈쭑쓽 湲곕뒫씠 젙긽씤 寃껋쑝濡 뙋떒뻽떎.

삉븳 젙옄쓽 깮議댁쑉, 誘명넗肄섎뱶由ъ븘 湲곕뒫 諛 썝삎吏덈쭑 寃궗瑜 쐞븯뿬 떆猷뚮쭏떎 理쒖냼 500媛 씠긽쓽 젙옄瑜 愿李고븯뿬 諛깅텇쑉濡 솚궛븯쑝硫, 蹂 떎뿕뿉꽌 뼸뼱吏 紐⑤뱺 寃곌낵뒗 Statistical Analysis System software version 9.2 (SAS Institute Inc. USA)瑜 씠슜븯뿬 遺꾩궛 遺꾩꽍쓣 떎떆븯쑝硫, Duncan’s multiple range test쓣 떎떆븯뿬 넻怨꾩쿂由щ 떎떆븯떎. 寃곌낵쓽 쑀쓽꽦 P<0.05 닔以뿉꽌 쑀쓽꽦쓣 寃젙븯떎.

洹 寃곌낵 Fig. 1뿉꽌 굹궃 諛붿 媛숈씠 젙옄쓽 솢젰 3 諛 9떆媛 룞븞 諛곗뼇븯쓣 븣 5 μM curcumin 泥④援ъ뿉꽌 쑀쓽쟻(P<0.05)쑝濡 利앷븳 寃껋쓣 솗씤븷 닔 엳뿀쑝硫, 5 μM curcumin씠 諛곗뼇떆媛 룞븞 젙옄쓽 깮議댁쑉쓣 쑀쓽쟻(P<0.05)쑝濡 利앷떆궎뒗 嫄 솗씤븯떎(Fig. 2). 씠肉먮쭔 븘땲씪 誘명넗肄섎뱶由ъ븘 솢꽦쓽 寃쎌슦 諛곗뼇떆媛꾧낵 愿怨 뾾씠 5 μM curcumin 泥섎━援ъ뿉꽌 쑀쓽쟻(P<0.05)쑝濡 利앷맂 寃껋쓣 솗씤븯떎(Fig. 3). 씪諛섏쟻쑝濡 curcumin 誘명넗肄섎뱶由ъ븘쓽 솢꽦쓣 뼢긽떆궗 肉먮쭔 븘땲씪 鍮꾩냼뿉 쓽븳 誘명넗肄섎뱶由ъ븘 룆꽦쓣 媛먯냼떆궓떎怨 븣젮졇 엳湲 븣臾몄뿉 curcumin뿉 쓽븳 뤌吏 젙옄쓽 誘명넗肄섎뱶由ъ븘 솢꽦 利앷뒗 씠쟾쓽 뿰援 寃곌낵 씪移섎맂떎怨 궗猷뚮맂떎(Muthumani et al., 2015; Zhang et al., 2015). 젙옄쓽 슫룞꽦뿉 씠슜릺뒗 ATP뒗 誘명넗肄섎뱶由ъ븘濡쒕꽣 깮꽦릺湲 븣臾몄뿉 誘명넗肄섎뱶由ъ븘 솢꽦 슫룞꽦怨 諛젒븳 뿰愿꽦쓣 媛吏怨 엳떎(Ruiz-Pesini et al., 1998; Sariozkan et al., 2009). 삉븳 誘명넗肄섎뱶由ъ븘뒗 꽭룷 궗硫몄쓣 議곗젅븯뒗 Bcl-2 떒諛깆쭏씠 쐞移섑븳 二쇱슂 냼湲곌쑝濡 븣젮졇 엳떎(Chen et al., 2015). 뵲씪꽌 curcumin 誘명넗肄섎뱶由ъ븘 솢꽦쓣 뼢긽떆耳 젙옄뿉 븘슂븳 슫룞꽦쓣 利앹쭊떆耳곗쓣 肉먮쭔 븘땲씪 꽭룷 궗硫몄쓣 뼲젣븯뿬 깮議댁쑉쓣 뼢긽떆耳곕떎怨 깮媛곷맂떎.

Fig. 1.

Effect of curcumin on boar sperm motility during liquid storage. Liquid semen was incubated in semen extender containing with 0, 5 and 10 μM curcumin for 9 hours, Bars represent means ± standard error of the mean (SEM). *Within the histogram, similarly shaded bars with different letters are significantly different (P<0.05)


Fig. 2.

Effect of curcumin on boar sperm viability during liquid storage. Liquid semen was incubated in semen extender containing with 0, 5 and 10 μM curcumin for 9 hours, and sperm viability was analyzed using Hoechst 33342 and propidium double staining assay. Bars represent means ± standard error of the mean (SEM). *Within the histogram, similarly shaded bars with different letters are significantly different (P<0.05)


Fig. 3.

Influence of curcumin on boar sperm mitochondrial activity during liquid storage. Liquid semen was incubated in semen extender containing with 0, 5 and 10 μM curcumin for 9 hours, and sperm mitochondrial was analyzed using rhodamine 123 and propidium double staining assay. Bars represent means ± standard error of the mean (SEM). *Within the histogram, similarly shaded bars with different letters are significantly different (P<0.05)


젙옄 꽭룷留 삩쟾꽦 Fig. 4뿉꽌 굹궃 諛붿 媛숈씠 諛곗뼇떆媛꾧낵 愿怨 뾾씠 5 μM curcumin뿉 쓽빐 쑀쓽쟻쑝濡 利앷릺뿀떎(P<0.05). 꽭룷留됱 꽭룷 궡遺 쇅遺瑜 援щ텇吏뼱 二쇰뒗 寃쎄퀎씤 룞떆뿉 꽭룷 궡쇅쓽 臾쇱쭏 닔넚씠굹 떊샇 쟾떖쓣 떞떦븯怨 엳뒗 以묒슂븳 湲곌씠떎(Shafiei et al., 2015). 룷쑀瑜섏뿉꽌 젙옄쓽 꽭룷留 吏吏 꽦遺꾩 泥댁꽭룷뒗 떎瑜닿쾶 씤吏吏덉쓣 룷븿븳 룷솕 諛 遺덊룷솕 吏諛⑹궛씠 留ㅼ슦 넂 븿웾쓣 蹂댁씠硫, 뵲씪꽌 怨쇰룄븳 솢꽦궛냼 끂異쒖뿉 듅엳 誘쇨컧븯떎(Kemal Duru et al., 2000; Sanocka et al., 2004). 삉븳 泥⑥껜 諛섏쓳 諛 젙옄 궃옄쓽 닔젙뿉 엳뼱 以묒슂븳 뿭븷쓣 븿쑝濡 닔젙 뒫젰뿉 븳 룊媛濡 옉슜븷 닔 엳떎(Shafiei et al., 2015). 蹂 떎뿕 寃곌낵뿉꽌 젙옄쓽 썝삎吏덈쭑 븞젙꽦씠 curcumin뿉 쓽빐 利앷맖쓣 솗씤븯怨, curcumin씠 떎뼇븳 異⑷꺽쑝濡쒕꽣 諛쒖깮븷 닔 엳뒗 썝삎吏덈쭑쓽 넀긽쓣 媛쒖꽑떆궗 닔 엳뒗 寃껋쓣 솗씤븯떎.

Fig. 4.

Effect of curcumin on boar sperm plasma membrane integrity during liquid storage. Liquid semen was incubated in semen extender containing with 0, 5 and 10 μM curcumin for 9 hours, and sperm plasma membrane integrity was analyzed using hypoosmotic swelling test. Bars represent means ± standard error of the mean (SEM). *Within the histogram, similarly shaded bars with different letters are significantly different (P<0.05)


씠윭븳 寃곌낵瑜 醫낇빀븯뿬 蹂댁븯쓣 븣 5 μM curcumin 뤌吏 젙옄쓽 슫룞꽦, 誘명넗肄섎뱶由ъ븘 솢룞꽦 諛 썝삎吏덈쭑 삩쟾꽦뿉 湲띿젙쟻씤 쁺뼢쓣 誘몄낀쑝굹 10 μM curcumin 泥섎━援ъ쓽 寃쎌슦 젙옄쓽 꽦긽씠 議곌뎄 李⑥씠媛 뾾嫄곕굹 삤엳젮 궙븘吏뒗 寃껋쑝濡 蹂댁븯쓣 븣 10 μM curcumin 뤌吏 젙옄뿉 쑀빐븳 쁺뼢쓣 誘몄튇떎怨 뙋떒맂떎. 씠윭븳 寃곌낵뒗 쟻젅븳 냽룄쓽 빆궛솕젣뒗 젙옄뿉 씠濡쒖슫 쁺뼢쓣 二쇱留, 怨쇰룄븳 냽룄쓽 빆궛솕젣뒗 젙옄쓽 湲곕뒫쓣 넀긽떆궓떎뒗 뿰援 寃곌낵 씪移섑븳떎怨 궗猷뚮맂떎(Kumar et al., 2015). 떎젣濡 怨좊냽룄쓽 curcumin 젙옄媛 븫而 깮떇湲곗씤 吏덉뿉꽌 옄沅곴꼍遺瑜 넻빐 옄沅곸뿉 뱾뼱媛湲 쟾뿉 젙옄瑜 二쎌뿬 궡洹 옉슜쓣 븯뒗 궡젙젣(Naz et al., 2014)뿉 궗슜릺湲 븣臾몄뿉, 10 μM怨 媛숈 怨좊냽룄쓽 curcumin쓽 뤌吏 젙옄뿉 엳뼱 븙 쁺뼢쓣 誘몄낀쓣 寃껋씠씪 깮媛곷맂떎. 떎젣濡 룷쑀룞臾 깮由ъ뿉 엳뼱 10 μM 씠긽쓽 curcumin뿉 븳 뿰援 寃곌낵(Naz et al., 2014; Rithaporn et al., 2003)瑜 諛뷀깢쑝濡 鍮꾩텛뼱 蹂댁븯쓣 븣, 蹂 떎뿕뿉꽌뒗 뤌吏 젙븸뿉꽌 curcumin쓽 빆궛솕젣濡 옉슜븷 닔 엳뒗 理쒖쟻쓽 냽룄뒗 10 μM 씠븯씪怨 뙋떒븯怨, 떎젣濡 10 μM curcumin뿉 鍮꾪븯뿬 5 μM curcumin씠 뤌吏 젙옄쓽 슫룞꽦, 깮議댁꽦, 誘명넗肄섎뱶由ъ븘 諛 썝삎吏덈쭑 삩쟾꽦뿉 씠濡쒖슫 쁺뼢쓣 二쇰뒗 寃껋쑝濡 굹궗떎.

뵲씪꽌 뤌吏 젙븸뿉 엳뼱 5 μM curcumin 9떆媛 룞븞 깮떇꽭룷씤 젙옄쓽 湲곕뒫쓣 뼢긽떆궗 닔 엳떎怨 뙋떒맂떎. 씠뒗 curcumin씠 떎뼇븳 궛솕 뒪듃젅뒪濡쒕꽣 諛쒖깮븷 닔 엳뒗 깮떇꽭룷 넀긽쓣 蹂댄샇븯뿬 룷쑀룞臾쇱쓽 踰덉떇 뒫젰쓣 뼢긽떆궗 닔 엳뒗 빆궛솕젣 젣젣濡 씠슜맆 닔 엳쓬쓣 굹궡怨 엳떎.

ACKNOWLEDGEMENTS

This study was supported by 2016 Research Grant from Kangwon National University (No. 520160128).

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