Effects of Saikosaponin on Antioxidant Capacity and Heat Shock Protein Gene Expression in Heat-Induced Bovine Mammary Epithelial Cells

HU Haitao; SUN Xianzhi; HUANG Feng; WANG Qingfeng; LI Jingjing; ZHUO Zhao; FAN Caiyun; SU Yanjing; CHENG Jianbo

doi:10.3969/j.issn.1006-267x.2021.11.041

2021 , Vol. 33 >Issue 11: 6420 - 6430

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.11.041

MOLECULAR AND CELLULAR NUTRITION

Effects of Saikosaponin on Antioxidant Capacity and Heat Shock Protein Gene Expression in Heat-Induced Bovine Mammary Epithelial Cells

HU Haitao ,
SUN Xianzhi ,
HUANG Feng ,
WANG Qingfeng ,
LI Jingjing ,
ZHUO Zhao ,
FAN Caiyun ,
SU Yanjing ,
CHENG Jianbo

Expand

1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China;
2. Shanghai Urban Construction Vocational College, Shanghai 201415, China;
3. Bright Animal Husbandry Co., Ltd., Shanghai 200436, China

Received date: 2021-04-16

Online published: 2021-11-10

Supported by

Fold

Abstract

This experiment was conducted to investigate the effects of saikosaponin-a (SSa) and saikosaponin-d (SSd) on antioxidant capacity and heat shock protein gene expression in heat-induced bovine mammary epithelial cells (BMECs). BMECs were cultured at 38℃ as negative control group; BMECs were cultured at 42℃ for 1, 4, 8, 12 and 24 h to establish a heat stress model (positive control group); BMECs medium was supplemented with 0.1, 1.0 μmol/L SSa and 0.01, 0.10 μmol/L SSd, and then heat induced at 42℃ for 1, 4, 8, 12 and 24 h (treatment groups). The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) in BMECs and the content of malondialdehyde (MDA) in the culture medium were measured of each group, and the mRNA relative expression levels of heat shock transcription factor-1 (HSF-1) and heat shock protein 70 (HSP70) in the cells were also measured. The results showed as follows:compared with the negative control group, the activities of SOD and GSH-Px in BMECs of the positive control group were significantly reduced (P<0.05), the content of MDA in the culture medium was significantly increased at 4 h of heat induction (P<0.05), and the mRNA relative expression levels of HSF-1 and HSP70 were significantly up-regulated (P<0.05). Compared with the positive control group, any concentration of SSa in the culture medium could significantly increase the SOD activity in BMECs at 1 h of heat induction (P<0.05); at 4, 8 and 24 h of heat induction, the CAT activity in BMECs of the 0.1 μmol/L SSa group was significantly increased (P<0.05); 0.01 μmol/L SSd could significantly increase the activity of GSH-Px in BMECs at 1, 4, 8, 12 and 24 h of heat induction (P<0.05); but at 12 and 24 h of heat induction, 1.0 μmol/L SSa and 0.10 μmol/L SSd did not significantly improve the activities of SOD, GSH-Px and CAT in BMECs (P>0.05), or significantly reduced the above antioxidant enzyme activities (P<0.05); the content of MDA in the culture medium of the 0.1 μmol/L SSd group was significantly increased at 12 h of heat induction (P<0.05), indicated that the relieving effect of SSa and SSd on oxidative stress of BMECs presented dose- and time-dependent manner. At 1 h of heat induction, the mRNA relative expression level of HSP70 in BMECs of the treatment groups was significantly upregulated compared with the positive control group (P<0.05); at 4 h of heat induction, the mRNA relative expression levels of HSF-1 and HSP70 in BMECs of the 0.1 μmol/L SSa-treated group was significantly upregulated compared with the positive control group (P<0.05), and at 12 and 24 h of heat induction, the mRNA relative expression level of HSP70 in BMECs of the 0.01 μmol/L SSd group was significantly up-regulated compared with the positive control group (P<0.05). In conclusion, heat treatment at 42℃ induces oxidative stress in BMECs, the proper concentrations of SSa and SSd alleviate heat-induced oxidative stress in BMECs, in which 0.1 μmol/L SSa and 0.01 μmol/L SSd have better alleviating effect on oxidative stress in BMECs, and also increase HSF-1 and HSP70 gene expression in BMECs under 42℃ heat-induced conditions, thus, BMECs are protected from cell damage caused by high temperature induction.

Key words： bovine mammary epithelial cells; heat induction; saikosaponin; oxidative stress; heat shock protein

Cite this article

HU Haitao , SUN Xianzhi , HUANG Feng , WANG Qingfeng , LI Jingjing , ZHUO Zhao , FAN Caiyun , SU Yanjing , CHENG Jianbo . Effects of Saikosaponin on Antioxidant Capacity and Heat Shock Protein Gene Expression in Heat-Induced Bovine Mammary Epithelial Cells[J]. Chinese Journal of Animal Nutrition, 2021 , 33(11) : 6420 -6430 . DOI: 10.3969/j.issn.1006-267x.2021.11.041

References

[1] FAN C Y, SU D, TIAN H, et al.Milk production and composition and metabolic alterations in the mammary gland of heat-stressed lactating dairy cows[J].Journal of Integrative Agriculture, 2019, 18(12):2844-2853.

[2] LI L, SUN Y, WU J, et al.The global effect of heat on gene expression in cultured bovine mammary epithelial cells[J].Cell Stress & Chaperones, 2015, 20(2):381-389.

[3] SAKATANI M, BALBOULA A Z, YAMANAKA K, et al.Effect of summer heat environment on body temperature, estrous cycles and blood antioxidant levels in Japanese black cow[J].Animal Science Journal, 2012, 83(5):394-402.

[4] HOLMSTRÖM K M, FINKEL T.Cellular mechanisms and physiological consequences of redox-dependent signalling[J].Nature Reviews.Molecular Cell Biology, 2014, 15(6):411-421.

[5] 权素玉, 张源淑, 卜登攀.热应激造成奶牛乳腺上皮细胞损伤并影响乳合成相关载体的基因表达[J].畜牧兽医学报, 2016, 47(8):1704-1713. QUAN S Y, ZHANG Y S, BU D P.Heat stress-induced cell injury and effects on the gene expression of milk synthesis-related transporters in dairy cow[J].Acta Veterinaria et Zootechnica Sinica, 2016, 47(8):1704-1713.(in Chinese)
[6] 胡菡, 王加启, 李发弟, 等.高温诱导体外培养奶牛乳腺上皮细胞的应激响应[J].农业生物技术学报, 2011, 19(2):287-293. HU H, WANG J Q, LI F D, et al.Responses of cultrued bovine mammary epithelial cells to heat stress[J].Journal of Agricultural Biotechnology, 2011, 19(2):287-293.(in Chinese)
[7] 恒景会, 田敏, 管武太, 等.氧化应激对母畜乳腺功能的影响及其营养调控策略[J].动物营养学报, 2020, 32(12):5587-5595. HENG J H, TIAN M, GUAN W T, et al.Effects of oxidative stress on mammary gland function of female animals and its nutritional regulation strategy[J].Chinese Journal of Animal Nutrition, 2020, 32(12):5587-5595.(in Chinese)
[8] WANG F G, ZHAO Y, CHEN S X, et al.Astragaloside Ⅳ alleviates ammonia-induced apoptosis and oxidative stress in bovine mammary epithelial cells[J].International Journal of Molecular Sciences, 2019, 20(3):600.
[9] 张云波, 梁园, 夏爱军.柴胡解热作用的药理研究进展[J].中国药业, 2011, 20(3):79-80. ZHANG Y B, LIANG Y, XIA A J.Progress in pharmacological research on antipyretic effect of Bupleurum[J].China Pharmaceuticals, 2011, 20(3):79-80.(in Chinese)
[10] ASHOUR M L, WINK M.Genus Bupleurum:a review of its phytochemistry, pharmacology and modes of action[J].Journal of Pharmacy and Pharmacology, 2011, 63(3):305-321.

[11] 薛燕.柴胡的解热作用药效学研究[J].中医药学刊, 2003, 21(11):1897-1897, 1959. XUE Y.Pharmacodynamics study on antipyretic effect of Bupleuri[J].Chinese Archives of Traditional Chinese Medicine, 2003, 21(11):1897-1897, 1959.(in Chinese)
[12] 金国泰, 李博, 王树荣.柴胡解热的物质基础、药效及机制研究[J].西部中医药, 2014, 27(2):20-22. JIN G T, LI B, WANG S R.Research on the mechanism, the effects and material foundation of Chaihu in relieving fever[J].Western Journal of Traditional Chinese Medicine, 2014, 27(2):20-22.(in Chinese)
[13] YUAN B C, YANG R, MA Y S, et al.A systematic review of the active saikosaponins and extracts isolated from Radix Bupleuri and their applications[J].Pharmaceutical Biology, 2017, 55(1):620-635.

[14] DANG S S, WANG B F, CHENG Y A, et al.Inhibitory effects of saikosaponin-d on CCl₄-induced hepatic fibrogenesis in rats[J].World Journal of Gastroenterology, 2007, 13(4):557-563.

[15] 余刘勤, 贾爱梅, 宋永砚.柴胡皂苷抗炎、抗氧化和降脂研究进展[J].中国动脉硬化杂志, 2020, 28(1):87-92. YU L Q, JIA A M, SONG Y Y.Progress in the study of saikosaponins on anti-inflammation, anti-oxidation and lipid-lowering effects[J].Chinese Journal of Arteriosclerosis, 2020, 28(1):87-92.(in Chinese)
[16] PAN L, BU D P, WANG J Q, et al.Effects of Radix Bupleuri extract supplementation on lactation performance and rumen fermentation in heat-stressed lactating Holstein cows[J].Animal Feed Science and Technology, 2014, 187:1-8.
[17] 范彩云, 苏娣, 侍宝路, 等.柴胡提取物对热应激奶牛生产性能和血液代谢的影响[J].草业科学, 2017, 34(12):2538-2545. FAN C Y, SU D, SHI B L, et al.Effects of Bupleurum extract on the performance and blood metabolism in heat-stressed dairy cows[J].Pratacultural Science, 2017, 34(12):2538-2545.(in Chinese)
[18] PADILLA L, MATSUI T, KAMIYA Y, et al.Heat stress decreases plasma vitamin C concentration in lactating cows[J].Livestock Science, 2006, 101(1/2/3):300-304.
[19] 陆灿强, 舒邓群, 臧一天.热应激诱导畜禽氧化应激、热休克反应与免疫和炎症反应的机制及营养调控措施[J].动物营养学报, 2021, 33(6):3115-3124. LU C Q, SHU D Q, ZANG Y T.Mechanism and nutritional regulation of oxidative stress, heat shock response, immunity and inflammation induced by heat stress in livestock and poultry[J].Chinese Journal of Animal Nutrition, 2021, 33(6):3115-3124.(in Chinese)
[20] FATTMAN C L, SCHAEFER L M, OURY T D.Extracellular superoxide dismutase in biology and medicine[J].Free Radical Biology & Medicine, 2003, 35(3):236-256.

[21] YANG L L, HUANG M S, HUANG C C, et al.The association between adult asthma and superoxide dismutase and catalase gene activity[J].International Archives of Allergy and Immunology, 2011, 156(4):373-380.

[22] MASELLA R, DI BENEDETTO R, VARÌ R, et al.Novel mechanisms of natural antioxidant compounds in biological systems:involvement of glutathione and glutathione-related enzymes[J].The Journal of Nutritional Biochemistry, 2005, 16(10):577-586.

[23] KONVIČNÁ J, VARGOVÁ M, PAULÍKOVÁ I, et al.Oxidative stress and antioxidant status in dairy cows during prepartal and postpartal periods[J].Acta Veterinaria Brno, 2015, 84(2):133-140.

[24] 付晶晶, 肖海芳, 宋元达.金银花等6种植物提取物总黄酮含量与抗氧化性相关性研究[J].食品与机械, 2017, 33(6):159-163. FU J J, XIAO H F, SONG Y D.Acomparative study of contents of total flavonoids and their antioxidant activities in six plants[J].Food & Machinery, 2017, 33(6):159-163.(in Chinese)
[25] AMORATI R, FOTI M C, VALGIMIGLI L.Antioxidant activity of essential oils[J].Journal of Agricultural and Food Chemistry, 2013, 61(46):10835-10847.

[26] WU S J, TAM K W, TSAI Y H, et al.Curcumin and saikosaponin a inhibit chemical-induced liver inflammation and fibrosis in rats[J].The American Journal of Chinese Medicine, 2010, 38(1):99-111.

[27] FAN J H, LI X, LI P, et al.Saikosaponin-d attenuates the development of liver fibrosis by preventing hepatocyte injury[J].Biochemistry and Cell Biology, 2007, 85(2):189-195.

[28] CHEN R J, GUO X Y, CHENG B H, et al.Saikosaponin a inhibits cigarette smoke-induced oxidant stress and inflammatory responses by activation of Nrf2[J].Inflammation, 2018, 41(4):1297-1303.

[29] FU Y H, HU X Y, CAO Y G, et al.Saikosaponin a inhibits lipopolysaccharide-oxidative stress and inflammation in human umbilical vein endothelial cells via preventing TLR4 translocation into lipid rafts[J].Free Radical Biology & Medicine, 2015, 89:777-785.
[30] 何燕, 胡志峰, 李平, 等.柴胡皂苷d抗肝纤维化大鼠脂质过氧化作用的研究[J].中国中药杂志, 2008, 33(8):915-919. HE Y, HU Z F, LI P, et al.Experimental study of saikosaponin-d (SSd) on lipid peroxidation of hepatic fibrosis on rat[J].China Journal of Chinese Materia Medica, 2008, 33(8):915-919.(in Chinese)
[31] LIN L B, QUE R Y, SHEN Y T, et al.Saikosaponin-d alleviates carbon-tetrachloride induced acute hepatocellular injury by inhibiting oxidative stress and NLRP3 inflammasome activation in the HL-7702 cell line[J].Molecular Medicine Reports, 2018, 17(6):7939-7946.
[32] 李素婷, 姜凌雪, 周晓慧, 等.柴胡皂苷-d对乙醇损伤原代培养大鼠肝细胞的保护作用及其机制研究[J].时珍国医国药, 2008, 19(11):2752-2753. LI S T, JIANG L X, ZHOU X H, et al.The protective effect and mechanisms of saikosaponin-d on primary cultured rats hepatocytes injured by ethanol[J].Lishizhen Medicine and Materia Medica Research, 2008, 19(11):2752-2753.(in Chinese)
[33] LIN X M, WU S D, WANG Q, et al.Saikosaponin-d reduces H₂O₂-induced PC12 cell apoptosis by removing ROS and blocking MAPK-dependent oxidative damage[J].Cellular and Molecular Neurobiology, 2016, 36(8):1365-1375.

[34] LI X J Y, LI X Y, HUANG N N, et al.A comprehensive review and perspectives on pharmacology and toxicology of saikosaponins[J].Phytomedicine, 2018, 50:73-87.
[35] 李晓宇, 尹利顺, 孙蓉.柴胡皂苷a对小鼠急性毒性实验研究[J].中国药物警戒, 2014, 11(8):449-452. LI X Y, YIN L S, SUN R.Experimental study on mice's acute toxicity of saikosaponin a[J].Chinese Journal of Pharmacovigilance, 2014, 11(8):449-452.(in Chinese)
[36] CHEN L, ZHANG F, KONG D S, et al.Saikosaponin d disrupts platelet-derived growth factor-β receptor/p38 pathway leading to mitochondrial apoptosis in human LO2 hepatocyte cells:a potential mechanism of hepatotoxicity[J].Chemico-Biological Interactions, 2013, 206(1):76-82.

[37] LIU M, ZHANG G F, NAQVI S, et al.Cytotoxicity of saikosaponin a targets HEKa cell through apoptosis induction by ROS accumulation and inflammation suppression via NF-κB pathway[J].International Immunopharmacology, 2020, 86:106751.
[38] WANG Q, ZHENG X L, YANG L, et al.Reactive oxygen species-mediated apoptosis contributes to chemosensitization effect of saikosaponins on cisplatin-induced cytotoxicity in cancer cells[J].Journal of Experimental & Clinical Cancer Research, 2010, 29(1):159.
[39] 黄伟, 吕征, 孙蓉.与功效和毒性相关的柴胡化学成分研究进展[J].中国药物警戒, 2010, 10(9):545-548. HUANG W, LV Z, SUN R.Research development on chemincal compositions in Bupleurum Chinense related with efficacy and toxicity[J].Chinese Journal of Pharmacovigilance, 2010, 10(9):545-548.(in Chinese)
[40] RICHTER K, HASLBECK M, BUCHNER J.The heat shock response:life on the verge of death[J].Molecular Cell, 2010, 40(2):253-266.

[41] HU H, ZHANG Y D, ZHENG N, et al.The effect of heat stress on gene expression and synthesis of heat-shock and milk proteins in bovine mammary epithelial cells[J].Animal Science Journal, 2016, 87(1):84-91.

[42] LI Q L, JU Z H, HUANG J M, et al.Two novel SNPs in HSF1 gene are associated with thermal tolerance traits in Chinese Holstein cattle[J].DNA and Cell Biology, 2011, 30(4):247-254.

[43] LEE W C, WEN H C, CHANG C P, et al.Heat shock protein 72 overexpression protects against hyperthermia, circulatory shock, and cerebral ischemia during heatstroke[J].Journal of Applied Physiology, 2006, 100(6):2073-2082.

[44] GUO S H, WHARTON W, MOSELEY P, et al.Heat shock protein 70 regulates cellular redox status by modulating glutathione-related enzyme activities[J].Cell Stress & Chaperones, 2007, 12(3):245-254.

[45] CHOI S, PARK K A, LEE H J, et al.Expression of Cu/Zn SOD protein is suppressed in hsp 70.1 knockout mice[J].Journal of Biochemistry and Molecular Biology, 2005, 38(1):111-114.
[46] SHEHATA A M, SAADELDIN I M, TUKUR H A, et al.Modulation of heat-shock proteins mediates chicken cell survival against thermal stress[J].Animals, 2020, 10(12):2407.
[47] ABDEL-MONEIM A M E, SHEHATA A M, ALZAHRANI S O, et al.The role of polyphenols in poultry nutrition[J].Journal of Animal Physiology and Animal Nutrition, 2020, 104(6):1851-1866.

[48] ZHANG B Z, GUO X T, CHEN J W, et al.Saikosaponin-d attenuates heat stress-induced oxidative damage in LLC-PK1 cells by increasing the expression of anti-oxidant enzymes and HSP72[J].The American Journal of Chinese Medicine, 2014, 42(5):1261-1277.

Options

Outlines

模态框（Modal）标题

Abstract

Cite this article

References