植物乳杆菌CGMCC8198破碎上清液对黑色素生成的抑制作用
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摘要
黑色素是由黑色素细胞合成的一种生物色素,能避免皮肤被紫外线灼伤,过度增长或分布不均会影响美观,导致老年斑、雀斑、黑斑病等病症.通过对植物乳杆菌(Lactobacillus plantarum)CGMCC8198破碎上清液(LAS)的抗氧化和黑色素生成抑制活性进行检测,结果表明:LAS能明显清除ABTS和DPPH自由基,并显著降低小鼠黑色素瘤细胞B16F10内黑色素的生成,且对细胞活性几乎无影响.LAS可降低细胞内的黑色素生成关键酶——酪氨酸酶的活性,而对细胞外酪氨酸酶的活性则无影响.通过实时荧光定量PCR(Real-timePCR)、免疫印迹实验(Westernblot)和荧光素酶报告基因检测分析,结果表明:LAS能够抑制酪氨酸酶和其相关蛋白家族成员TRP-1、TRP-2及该家族上游关键转录调控因子小眼畸形相关转录因子(MITF)的转录表达;抑制MITF以及TYR启动子转录活性.
Melanin is a biological pigment synthesized by melanocytes,which can prevent the skin from being burned by UV rays. Overgrowth or uneven distribution of melanin can affect the appearance and lead to senile plaques,freckles,black spots and other diseases. The antioxidization ability and melanogenesis inhibition of the cell lysis supernatant(LAS)prepared from Lactobacillus plantarum CGMCC8198 were tested. LAS can clearly scavenge free radicals ABTS and DPPH. In B16 F10 melanoma cells,LAS significantly reduced the intracellular melanin production,but did not affect the cell activity.Then,the effects of LAS on the activity of tyrosine and the key enzyme of melanin production were analyzed. LAS reduced the activity of intracellular tyrosinase and had no effect on the activity of extracellular tyrosinase. The effect of LAS was analyzed by RT-PCR,Western blot and Luciferase reporter assay. LAS could inhibit not only the transcription and expression of tyrosinase,tyrosinase-related protein TRP-1 and TRP-2,but also their key regulation factor MITF. The results of luciferase reporter assay show that LAS could inhibit the promoter activity of both TYR and MITF.
Melanin is a biological pigment synthesized by melanocytes,which can prevent the skin from being burned by UV rays. Overgrowth or uneven distribution of melanin can affect the appearance and lead to senile plaques,freckles,black spots and other diseases. The antioxidization ability and melanogenesis inhibition of the cell lysis supernatant(LAS)prepared from Lactobacillus plantarum CGMCC8198 were tested. LAS can clearly scavenge free radicals ABTS and DPPH. In B16 F10 melanoma cells,LAS significantly reduced the intracellular melanin production,but did not affect the cell activity.Then,the effects of LAS on the activity of tyrosine and the key enzyme of melanin production were analyzed. LAS reduced the activity of intracellular tyrosinase and had no effect on the activity of extracellular tyrosinase. The effect of LAS was analyzed by RT-PCR,Western blot and Luciferase reporter assay. LAS could inhibit not only the transcription and expression of tyrosinase,tyrosinase-related protein TRP-1 and TRP-2,but also their key regulation factor MITF. The results of luciferase reporter assay show that LAS could inhibit the promoter activity of both TYR and MITF.
引文
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[23]Ji K,Cho Y S,Kim Y T.Tyrosinase inhibitory and antioxidative effects of lactic acid bacteria isolated from dairy cow feces[J].Probiotics and Antimicrobial Proteins,2011,83(3):980-987.
[24]杨越楠,王震宇,商进.L-抗坏血酸在黑素合成中的作用[J].中国现代医生,2016,54(27):166-168.
[25]杨英歌,黄继翔,马凯,等.抑制黑色素合成的乳酸菌胞外多糖的筛选和性质研究[J].微生物学通报,2017,44(3):583-590.
[2]Sima V H,Patris S,Aydogmus Z,et al.Tyrosinase immobilized magnetic nanobeads for the amperometric assay of enzyme inhibitors:Application to the skin whitening agents[J].Talanta,2011,83(3):980-987.
[3]Briganti S,Camera E,Picardo M.Chemical and instrumental approaches to treat hyperpigmentation[J].Pigment Cell&Melanoma Research,2003,16(2):101-110.
[4]姚彬,张莉莉,任清.皮肤黑色素的形成及美白剂美白机理[J].北京日化,2007,12(2):18-22.
[5]Draelos Z D.Skin lightening preparations and the hydroquinone controversy[J].Dermatologic Therapy,2007,20(5):308-313.
[6]陈永红,邹志飞,王岚,等.曲酸对动物的毒性研究及安全性评价[J].食品科学,2007,28(9):536-540.
[7]Solano F,Briganti S,Picardo M,et al.Hypopigmenting agents:An updated review on biological,chemical and clinical aspects[J].Pigment Cell&Melanoma Research,2006,19(6):550-571.
[8]Lin M Y,Yen C L.Antioxidative ability of lactic acid bacteria[J].Journal of Agricultural&Food Chemistry,1999,47(4):1460-1466.
[9]Chakraborty A K,Funasaka Y,Slominski A,et al.Production and release of proopiomelanocortin(POMC)derived peptides by human melanocytes and keratinocytes in culture:Regulation by ultraviolet B[J].Biochimica Et Biophysica Acta,1996,1313(2):130-138.
[10]Uraiwan P,Vanida T,Tasanee O,et al.Inhibition of UVA-mediated melanogenesis by ascorbic acid through modulation of antioxidant defense and nitric oxide system[J].Archives of Pharmacal Research,2011,34(5):811-820.
[11]Quevedo W C,Holstein T J,Dyckman J,et al.Inhibition of UVR-induced tanning and immunosuppression by topical applications of vitamins C and E to the skin of hairless(hr/hr)mice[J].Pigment Cell Research,2000,13(2):89-98.
[12]张江巍,曹郁生,李海星,等.乳酸菌抗氧化活性及检测方法[J].中国乳品工业,2005,33(9):53-56.
[13]Lin M Y,Chang F J.Antioxidative effect of intestinal bacteria Bifidobacterium longum ATCC15708 and Lactobacillus acidophilus ATCC4356[J].Digestive Diseases&Sciences,2000,45(8):1617-1622.
[14]Kaizu H,Sasaki M,Nakajima H,et al.Effect of antioxidative lactic acid bacteria on rats fed a diet deficient in vitamin E[J].Journal of Dairy Science,1993,76(9):2493-2499.
[15]Liu C F,Pan T M.In vitro effects of lactic acid bacteria on cancer cell viability and antioxidant activity[J].Journal of Food&Drug Analysis,2010,18(2):77-86.
[16]Kullisaar T,Songisepp E,Mikelsaar M,et al.Antioxidative probiotic fermented goats'milk decreases oxidative stress-mediated atherogenicity in human subjects[J].British Journal of Nutrition,2003,90(2):449-456.
[17]Talwalkar A,Kailasapathy K.Metabolic and biochemical responses of probiotic bacteria to oxygen[J].Journal of Dairy Science,2003,86(8):2537-2546.
[18]张同存,罗学刚,古向超,等.高耐胆盐能力的菌株及胆盐水解酶基因:ZL201310491340.4[P].2015-12-23.
[19]张同存,罗学刚,古向超.植物乳杆菌在降血脂和辅助减肥方面的作用:ZL201310518617.8[P].2015-05-20.
[20]Jang J Y,Kim H N,Kim Y R,et al.Aqueous fraction from Cuscuta japonica seed suppresses melanin synthesis through inhibition of the p38 mitogen-activated protein kinase signaling pathway in B16F10 cells[J].Journal of Ethnopharmacology,2012,141(1):338-344.
[21]Kim H R,Kim H,Jung B J,et al.Lipoteichoic acid isolated from Lactobacillus plantarum inhibits melanogenesis in B16F10 mouse melanoma cells[J].Molecules&Cells,2015,38(2):163-170.
[22]冯荣楷,唐海谊,周志敏.美白酪氨酸酶和腺苷酸环化酶抑制剂的计算机毒性研究[J].计算机与应用化学,2012,29(10):1231-1236.
[23]Ji K,Cho Y S,Kim Y T.Tyrosinase inhibitory and antioxidative effects of lactic acid bacteria isolated from dairy cow feces[J].Probiotics and Antimicrobial Proteins,2011,83(3):980-987.
[24]杨越楠,王震宇,商进.L-抗坏血酸在黑素合成中的作用[J].中国现代医生,2016,54(27):166-168.
[25]杨英歌,黄继翔,马凯,等.抑制黑色素合成的乳酸菌胞外多糖的筛选和性质研究[J].微生物学通报,2017,44(3):583-590.