• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Effects of Bupleurum extract on blood metabolism, antioxidant status and immune function in heat-stressed dairy cows

    2018-03-07 11:39:59CHENGJianboFANCaiyunSUNXianzhiWANGJiaqiZHENGNanZHANGXingkaiQlNJunjieWANGXiumin
    Journal of Integrative Agriculture 2018年3期

    CHENG Jian-bo, FAN Cai-yun, SUN Xian-zhi, WANG Jia-qi, ZHENG Nan, ZHANG Xing-kai, QlN Jun-jie, WANG Xiu-min

    1 College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, P.R.China

    2 Laboratory of Quality and Safety Risk Assessment for Dairy Products of Ministry of Agriculture, Institute of Animal Science,Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China

    3 State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193,P.R.China

    4 Shanghai Bright Holstan Co., Ltd., Shanghai 200436, P.R.China

    5 Beijing Centre Biology Co., Ltd., Beijing 102206, P.R.China

    1.lntroduction

    Heat stress not only reduces production performance(Collier et al.2006; Hammami et al.2012) but also has adverse effects on the health status of dairy cows.These adverse effects include an increased incidence of metabolic disorders (Wheelock et al.2010), in addition to increased oxidant stress (Lin et al.2006) and impaired immune function(Carroll et al.2012).Although many nutrition strategies,such as increasing the energy density of diets (Wang et al.2010) and rumen-protected niacin supplementation(Zimbelman et al.2013), have been used to alleviate heat stress, heat stress remains a costly issue for the global dairy industry (St-Pierre et al.2003).Consequently, new strategies need to be developed that can help maintain dairy cows’ health during hot summers.

    The genus Bupleurum, a major ingredient of oriental folk medicine, has been widely used in Asia for the treatment of many diseases over the past 2 000 years (Ashour and Wink 2011).It contains many secondary metabolites,such as polysaccharides (Sun et al.1991), saikosaponins(Navarro et al.2001), and essential oil (Martine et al.1993).Saikosaponin constitutes the main class of secondary metabolites in the genus Bupleurum, amounting for up to 7% of roots (Ashour and Wink 2011).Plants belonging to the genus Bupleurum have various pharmacological functions, with studies reporting antipyretic properties(Huang 1993), anti-inflammatory activity in rats (Martin et al.1993), antioxidant activity in rats (Zhao et al.2012),and immunostimulant activity in mice (Sun 2006).Previous research on Bupleurum has focused on humans and mice,whereas there has been little research on the effect of Bupleurum supplementation on cows.Our previous study suggested that Bupleurum extract (BE) could mitigate negative effects of heat stress on production in lactating Holstein cows (Pan et al.2014), but the effects of direct supplementation with BE on blood metabolites, antioxidant status, and immune responses in heat-stressed lactating Holstein cows are unknown.Therefore, the objective of this study was to investigate the effects of oral administration of BE on blood metabolites, antioxidant status, and immune function of lactating Holstein cows subjected to heat stress.

    2.Material and methods

    2.1.Animals, diet and experimental design

    All animals included in this study were maintained according to the principles of the Animal Care and Use Committee of the Chinese Academy of Agricultural Sciences.Forty lactating Holstein cows, (37.5±1.8) kg of milk d-1, (75±15) d in milk, and (1.7±0.4) parity, were categorized according to average daily milk yield, parity, and day in milk and randomly assigned to 1 of the following 4 treatments (n=10):0 (control), 0.25, 0.5, or 1.0 g of BE kg-1dry matter.A commercially available BE product was used (Beijing Centre Biology Co., Ltd., China) containing 35.0% BE (a mixture of 6.6% saikosaponins, 2.5% essential oil, and 25.9%polysaccharides) and 65.0% starch mix.In dairy cows, heat stress occurs when the temperature-humidity index (THI)exceeds 72 (Armstrong 1994).During the experiment, the mean THI in the barn in which the animals were housed was 78.2 (range: 71.9 to 80.8) at 06:00 h, 79.7 (range: 72.7 to 83.3) at 14:00 h, and 78.3 (range: 70.2 to 81.7) at 22:00 h(Pan et al.2014).

    The basal diet (Table 1) was formulated to meet or exceed nutrient recommendations (NRC 2001).Feedstuffs were offered (roughage first and then concentrated feed) daily at 05:30, 13:00, and 20:00 h to ensure 5% refusals, and the BE supplements were top dressed in equal portions to concentrated feed in the morning feeding.All the cows were assigned to an individual feeding column cangue,with access to fresh water.The experimental period lasted 10 weeks.

    2.2.Sampling procedure

    Blood samples were collected from the tail vein of each cow before the morning feed on days 0, 21, 42, and 63 of the experimental period.Blood samples were collected in EDTA-treated tubes for the analysis of CD4+and CD8+T lymphocytes.Additional blood samples were collected inserum tubes and centrifuged at 3 000×g for 10 min at 4°C.The serum samples were then stored at -40°C until analysis of the blood metabolites, antioxidant enzyme activities, and immune index.

    Table 1 Ingredients and chemical composition of basal diets(dry matter (DM) basis)

    2.3.Analytical procedures

    Blood metabolites were analyzed at 37°C using an automatic biochemistry analyzer (SYNCHRON CX5 PRO; Beckman Coulter, Fullerton, CA, USA).Glucose (GLU), triglyceride(TG), total cholesterol (TC), total protein (TP), albumin(ALB), blood urea nitrogen (BUN), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol(LDL-C) were analyzed using commercial kits (Instrumentation Laboratory, Lexington, MA, USA).Nonesterified fatty acid(NEFA) content was also analyzed using a commercial kit(Sekisui Medical Technology Ltd., Beijing, China).

    Serum superoxide dismutase (SOD) activity, total antioxidant capacity (T-AOC), glutathione peroxidase(GSH-Px) activity, and malondialdehyde (MDA) content were measured using standard kits (Jiancheng Biology Co.,Nanjing, China) according to the manufacturer’s instructions.

    CD4+and CD8+T lymphocytes were measured using commercial kits (AbD Serotec, USA) according to the method described by Caraher et al.(2000), using a Cell Lab QuantaTMSC Flow Cytometer (Beckman Coulter, Fullerton,CA, USA).The levels of immunoglobulin (Ig) A, IgM, and IgG were determined using Cow IgA (catalogue no.E10-121), IgM (catalogue no.E10-101), and IgG (catalogue no.E11-118) Enzyme-Linked Immunosorbent Assay (ELISA)Kits (Bethyl Laboratories, Montgomery, TX, USA).Briefly,capture antibody was used to coat a 96-well microtiter plate (BioFil, Canada) for 1 h at room temperature, and then ELISA wash solution containing 0.05% Tween-20 and 50 mmol L-1of Tris buffered saline was added for 0.5 h to block free binding sites.Next, duplicate serum samples were diluted in ELISA wash solution and incubated for 1 h at room temperature.The plates were then incubated with IgA, IgM, and IgG horseradish peroxidase detection antibody for 1 h.After washing 5 times with ELISA wash solution, the plates were developed with tetramethylbenzidine for 15 min in the dark, and absorbance was then measured at 450 nm.Interleukin (IL)-2, IL-4, and IL-6 levels were determined using Cow ELISA Kits (Groundwork Biotechnology Diagnosticate,CA, USA).The tumor necrosis factor-α (TNF-α) level was measured using a Cow Radioimmunoassay Kit (North Institute of Biotechnology, Beijing, China) according to the manufacturer’s instruction.

    2.4.Statistical analysis

    Data on blood metabolic profiles, antioxidative enzyme activities, immunoglobulins, and cytokines were analyzed by repeated measures using the MIXED model procedure of SAS (2008).The statistical model contained treatment, time,and treatment×time interactions.Differences among the treatments were analyzed using Tukey’s multiple range test.Data on CD4+and CD8+T lymphocytes and the CD4+/CD8+ratio were analyzed using a GLM.Means are presented with the standard error of the mean in tabular form.A value of P<0.05 was considered statistically significant, P<0.01 was considered markedly significant, and P<0.1 was considered a tendency.

    3.Results

    3.1.Energy, lipid and protein metabolism in blood

    The effects of BE supplementation on blood metabolites are shown in Table 2.Compared with the controls, cows supplemented with BE had higher (P<0.05) concentrations of TP, whereas there were no differences (P>0.05) in GLU,NEFA, TG, TC, LDL-C, and HDL-C among the treatments.Cows fed 0.25 g of BE kg-1had increased (P<0.05) ALB levels compared with control cows, and the ALB level also tended to be higher (P<0.10) in cows fed 0.5 g of BE kg-1.In addition, cows fed 0.25 and 0.5 g of BE kg-1had decreased (P<0.05) BUN concentrations, and a lower tendency (P<0.10) toward BUN concentrations was found in cows fed 1.0 g of BE kg-1.

    3.2.Serum antioxidant enzyme activities

    SOD activity was elevated (P<0.05) by BE supplementation.GSH-Px activity tended to be higher (P<0.10) in cows fed 0.25 g of BE kg-1compared with the controls.GSH-Px activity was also higher (P=0.05) in cows fed 0.5 and 1.0 g of BE kg-1.Supplementation with BE had no effect (P>0.05)on serum T-AOC and MDA content.GSH-Px, T-AOC, and MDA showed significant time-dependent effects (P<0.05),as shown in Table 3.

    3.3.Serum CD4+ and CD8+ T lymphocyte levels and CD4+/CD8+ ratio

    Compared with the controls, BE supplementation had no effect (P>0.05) on the proportion of CD4+and CD8+T lymphocytes or the CD4+/CD8+ratio (Table 4).

    3.4.Serum antibody and cytokine levels

    The serum Ig and cytokine concentrations are shown in Table 5.Supplementation with BE had no effect (P>0.05)on the IgM concentration.However, IgA levels tendedto be higher (P<0.10) in cows fed 0.25 and 1.0 g of BE kg-1compared with the controls, and they were increased(P=0.01) in cows supplemented with 0.5 g of BE kg-1.Compared with the controls, concentrations of IgG and IL-2 were increased (P<0.05) in cows supplemented with 0.25 and 0.5 g of BE kg-1, and they tended to be higher (P<0.10)in cows fed 1.0 g of BE kg-1.Cows fed 0.5 and 1.0 g of BE kg-1had increased (P<0.05) IL-4 concentrations.IL-4 levels also tended to be higher (P<0.10) in cows fed 0.25 g of BE kg-1.Cows fed 0.25 g of BE kg-1had increased (P<0.05)IL-6 concentrations.A tendency (P<0.10) toward higher IL-6 concentrations was also found in cows fed 0.5 g of BE kg-1, whereas BE supplementation had no effect (P>0.05)on TNF-α levels.There were significant time-dependent changes (P<0.01) in IgG and IL-4 levels.

    Table 2 Effects of different dose of Bupleurum extract (BE) on blood metabolites related to energy, lipid and protein metabolism in heat-stressed dairy cows

    Table 3 Effects of different dose of Bupleurum extract (BE) on the activities of serum antioxidant enzymes in heat-stressed dairy cows

    Table 4 Effects of different dose of Bupleurum extract (BE) on the proportion of T lymphocyte subsets and the CD4+/CD8+ ratio in heat-stressed dairy cows

    4.Discussion

    Several studies demonstrated that heat stress reduced plasma GLU, cholesterol, TG, NEFA, and TP (Abeni et al.2007; Wheelock et al.2010), as well as causing oxidative stress (Bernabucci et al.2002) and reducing plasma antioxidant activity in dairy cows (Harmon et al.1997).Our previous study suggested that BE supplementation appeared to mitigate heat stress in lactating Holstein cows(Pan et al.2014).There have been no previous studies of the effect of BE supplementation on blood metabolism of heat-stressed cows.The present experiment showed that GLU, NEFA, TG, TC, LDL-C, and HDL-C did not differ among the treatments, suggesting that feeding BE had no effect on GLU and lipid metabolism.However, in the experiment,cows fed BE, especially 0.25 or 0.5 g kg-1had increased TP and ALB levels and decreased BUN concentrations.Thus, supplementation with BE appeared to improve nitrogen metabolism in heat-stressed dairy cows.In general, most plasma protein is synthesized by the liver, and the protein content is decreased when liver cells are damaged.The elevated blood protein levels found in the present study may be due to BE supplementation alleviating liver cell damage (Zhao et al.2012).Heat stress is known to increase BUN levels in dairy cows (Kamiya et al.2006).In the present study, the decrease in BUN levels induced by BE supplementation might be due to the alleviation of heat stress in dairy cows (Pan et al.2014).

    ?

    In this study, BE supplementation significantly increased the activities of SOD and GSH-Px, providing convincing evidence that BE can improve the antioxidant status of lactating cows under heat stress.A previous study reported that BE contained a high total phenol content, which inhibited lipid peroxidation and removed free radicals (Wang et al.2005).Zhao et al.(2012) found that SOD activity was elevated following treatment with a soluble polysaccharide fraction from Bupleurum chinese in a rat model of hepatic injury caused by D-galactosamine.Furthermore,purified polysaccharide isolated from Bupleurum chinense showed significant scavenging activity against free radicals (Sun et al.2010).In the present study, BE supplementation had no effect on the serum T-AOC and MDA content.This may be explained by the study animal or the BE supplemental level.In this study, activities of GSH-Px and T-AOC were higher in weeks 6 and 10 than in week 1, and MDA actavity was lower in week 10 than in week 1.The increase in antioxidant levels over time may explained by a decrease in the degree of heat stress in the dairy cows (Pan et al.2014).

    Previous research reported that heat stress inhibited immune function in cows (Caroprese et al.2009).Orally administered Bupleurum markedly promoted lymphocyte proliferation, enhanced the antibody titer (Wang et al.2005; Sun et al.2006), and increased cytokine and Ig secretion (Guo et al.2000).In the present study, serum IgA and IgG concentrations were statistically increased in cows fed 0.25 or 0.5 g of BE kg-1compared with the controls, suggesting that BE acts as an immune stimulant and enhances humoral immunity in heat-stressed dairy cows.This finding was consistent with that of Matsumoto et al.(2010), who observed that Hochuekkito, a Kampo (traditional Japanese herbal) medicine, enhanced mucosal IgA antibody responses in mice.Moreover, Yen et al.(1995) reported that serum levels of IgA, IgG,and IgM increased following treatment with a Bupleurum-derived saikosaponin in mice.However, supplementation with BE had no effect on IgM levels in the present study, which was not consistent with the findings of a previous study of mice (Guo et al.2000).The difference may be attributed to differences in breeds.

    CD4+and CD8+T lymphocytes are surface markers of T lymphocytes,which reflect the body’s cellular immune state (Park et al.1993).Currently,there is no direct evidence for effects of BE on CD4+T lymphocytes.However, one in vitro study showed that Sho-saiko-to (a Chinese herbal decoction containing BE, known as So-shi-ho-tang in Japanese and Xiao Chai-hu-tang in Chinese) had no effect on CD4+T lymphocyte counts in cultured splenocytes and hepatic mononuclear cells (Ohtake et al.2005),which was similar to that found in the present study.In our study, BE supplementation had no effect on the proportion of CD8+T lymphocytes, indicating that BE did not exert cytotoxic effects on cellular immunity.

    In the present study, BE supplementation increased the level of IL-2, which was in accordance with that found in a previous study by Liu et al.(1997), who observed in vitro that Xiao Chai-hu-tang, a BE water soluble substance,enhanced IL-2 secretion.The increased IL-2 levels in the BE-supplemented groups may be caused by T-lymphocyte proliferation (Malek 2003).Studies also reported that saikosaponin from Bupleurum augmented antibody formation (Yamaguchi et al.1985) and that it significantly increased IL-2 levels in mice (Yen et al.1995).An in vivo study showed that Sho-saiko-to significantly increased the IL-4 content in splenocytes of mice (Kang et al.2009),which was similar to the findings in our study.The increased IL-4 content found in the present study indicated that BE enhanced humoral and cellular immune responses in heatstressed dairy cows.Previous research reported that IL-6 contributed to the enhancement of nonspecific Ig secretion from B cells (Sakurai et al.1999).Guo et al.(2000) reported that bupleuran 2IIc (a pectic polysaccharide from the roots of Bupleurum falcatum) induced IL-6 secretion in normal murine B cells and in some specific B-cell lines in mice.In the present study, the IL-6 content was significantly increased in cows fed 0.25 g of BE kg-1, suggesting the BE promoted the differentiation of B lymphocytes.

    Overall, the results of the present study indicated that BE supplementation stimulated the secretion of serum immunoglobulins and cytokines, including IgA, IgM, IgG,IL-2, IL-4, and IL-6, thereby helping to enhance immune function.The enhanced immune status and function of dairy cows supplemented with BE in this study might be due to the alleviation of heat stress (Pan et al.2014), as well as that some active ingredients in Bupleurum transport to lymphoid tissue and subsequent interactions with lymphocytes (Yen et al.1995).Alternatively, polysaccharides from Bupleurum may have enhanced binding of immune complexes to peritoneal macrophages through Fc receptor expression(Matsumoto et al.1993).

    5.Conclusion

    The present study demonstrated that feeding BE to heatstressed dairy cows improved protein metabolism by increasing blood TP and ALB levels.It increased antioxidant activity by upregulating the activities of SOD and GSH-Px and enhanced immune function by increasing IgA, IgG,IL-2, and IL-4 concentrations.This study points to a new strategy to help mitigate negative effects of heat stress in lactating Holstein cows.

    Acknowledgements

    This study was supported financially by the National Key Research and Development Program of China(2016YFD0500503), the Agricultural Science and Technology Innovation Program, China (ASTIP-IAS12) and the Anhui International Science and Technology Cooperation Plan Program, China (1503062019).

    Abeni F, Calamari L, Stefanini L.2007.Metabolic conditions of lactating Friesian cows during the hot season in the Po valley.1.Blood indicators of heat stress.International Journal of Biometeorology, 52, 87-96.

    Armstrong D V.1994.Heat stress interaction with shade and cooling.Journal of Dairy Science, 77, 2044-2050.

    Ashour M L, Wink M.2011.Genus Bupleurum: A review of its phytochemistry, pharmacology and modes of action.Journal of Pharmacy and Pharmacology, 63, 305-321.

    Bernabucci U, Ronchi B, Lacetera N, Nardone A.2002.Markers of oxidative status in plasma and erythrocytes of transition dairy cows during hot season.Journal of Dairy Science,85, 2173-2179.

    Caraher E M, Parenteau M, Gruber H, Scott F W.2000.Flow cytometric analysis of intracellular IFN-γ, IL-4 and IL-10 in CD3+4+T-cells from rat spleen.Journal of Immunological Methods, 244, 29-40.

    Caroprese M, Marzano A, Entrican G, Wattegedera S,Albenzio M, Sevi A.2009.Immune response of cows fed polyunsaturated fatty acids under high ambient temperatures.Journal of Dairy Science, 96, 2796-2803.

    Carroll J A, Burdick N C, Chase J C C, Coleman S W, Spiers D E.2012.Influence of environmental temperature on the physiological, endocrine, and immune responses in livestock exposed to aprovocative immune challenge.Domestic Animal Endocrinology, 43, 146-153.

    Collier R J, Dahl G E, VanBaale M J.2006.Major advances associated with environmental effects on dairy cattle.Journal of Dairy Science, 89, 1244-1253.

    Guo Y J, Matsumoto T, Kikuchi Y J, Ikejima T, Wang B X,Yamada H.2000.Effects of a pectic polysaccharide from a medicinal herb, the roots of Bupleurum falcatum L.on interleukin 6 production of murine B cells and B cell lines.Immunopharmacology, 49, 307-316.

    Hammami H, Bormann J, M’hamdi N, Montaldo H H, Gengler N.2012.Evaluation of heat stress effects on production traits and somatic cell score of Holsteins in a temperate environment.Journal of Dairy Science, 96, 1-12.

    Harmon R J, Lu M, Trammel D S, Smith B A.1997.Influence of heat stress and calving on antioxidant activity in bovine blood.Journal of Dairy Science, 80(Suppl.1), 264.

    Huang K.1993.Antipyretic herbs.In: The Pharmacology of Chinese Herbs.CRC Press, Tokyo.pp.151-152.

    Kamiya M, Kamiya Y, Tanaka M, Oki T, Nishiba Y, Shioya S.2006.Effects of high ambient temperature and restricted feed intake on urinary and plasma 3-methylhistidine in lactating Holstein cows.Animal Science Journal, 77,201-207.

    Kang H, Choib T W, Ahnb K S, Leec J Y, Hamc I H, Choic H Y,Shim E S, Sohn N W.2009.Upregulation of interferon-γ and interleukin-4, Th cell-derived cytokines by So-Shi-Ho-Tang(Sho-Saiko-To) occurs at the level of antigen presenting cells, but not CD4+T cells.Journal of Ethnopharmacology,123, 6-14.

    Lin H, Decuypere E, Buyse J.2006.Acute heat stress induces oxidative stress in broiler chickens.Comparative Biochemistry and Physiology (Part A), 144, 11-17.

    Liu J J, Xue H, Chen Z F.1997.The effects of water soluble substance extracted from Xiao Chai-hu-tang on IL-2 production and CD antigen change of PHA-stimulated PBMC.Journal of Luzhou Medical College, 20, 331-334.(in Chinese)

    Malek T R.2003.The main function of IL-2 is to promote the development of T regulatory cells.Journal of Leukocyte Biology, 74, 961-965.

    Martin S, Padilla E, Ocete M A, Galvez J, Jiménez J, Zarzuelo A.1993.Anti-inflammatory activity of the essential oil of Bupleurum fruticescens.Planta Medica, 59, 533-536.

    Matsumoto T, Cyong J C, Kiyohara H, Matsui H, Abe A,Hirano M, Danbara H, Yamada H.1993.The pectic polysaccharides from Bupleurum falcatum L.enhances immune-complexes binding to peritoneal macrophages through Fc receptor expression.International Journal of Immunopharmacology, 15, 683-693.

    Matsumoto T, Noguchi M, Hayashi O, Makino K, Yamada H.2010.Hochuekkito, a kampo (traditional Japanese herbal)medicine, enhances mucosal IgA antibody response in mice immunized with antigen-entrapped biodegradable microparticles.Evidence-Based Comple and Alternative Medicine, 7, 69-77.

    Navarro P, Giner R M, Recio M C, Má?ez S, Cerdá-Nicolás M,Ríos J L.2001.In vivo anti-inflammatory activity of saponins from Bupleurum rotundifolium.Life Science, 68, 1199-1206.

    NRC (National Research Council).2001.Nutrient Requirements of Dairy Cattle.7th ed.National Academy Press, Washington,D.C.

    Ohtake N, Yamamoto M, Takeda S C, Aburada M, Ishige A, Watanabe K, Inoue M.2005.The herbal medicine Sho-saiko-to selectively inhibits CD8+T-cell proliferation.European Journal of Pharmacology, 507, 301-310.

    Pan L, Bu D P, Wang J Q, Cheng J B, Sun X Z, Zhou L Y, Qin J J, Zhang X K, Yuan Y M.2014.Effects of Radix Bupleuri extract supplementation on lactation performance and rumen fermentation in heat-stressed lactating Holstein cows.Animal Feed Science and Technology, 187, 1-8.

    Park Y H, Fox L K, Hamilton M J, Davis W C.1993.Suppression of proliferative response of BoCD4+T lymphocytes by activated BoCD8+T lymphocytes in the mammary gland of cows with Staphylococcus aureus mastitis.Veterinary Immunology and Immunopathology, 36, 137-151.

    Sakurai M H, Matsumoto T, Kiyohara H, Yamada H.1999.B cell proliferation activity of pectic polysaccharide from amedicinal herb, the roots of Bupleurum falcatum L.and its structural requirement.Immunology, 97, 540-547.

    SAS (Statistical Analysis System).2008.User’s Guide:Statistics.Version 9.2 edition.SAS Institute, Cary, NC.

    St-Pierre N R, Cobanov B, Schnitkey G.2003.Economic losses from heat stress by stressby US livestock industries.Journal of Dairy Science, 86(Suppl.), E52-E77.

    Sun H X.2006.Haemolytic activities and adjuvant effect of Bupleurum chinense saponins on the immune responses to ovalbumin in mice.Vaccine, 24, 1324-1331.

    Sun L W, Feng K, Jiang R, Chen J Q, Zhao Y, Ma R, Tong H B.2010.Water-soluble polysaccharide from Bupleurum chinense DC: Isolation, structural features and antioxidant activity.Carbohydrate Polymers, 79, 180-183.

    Sun X B, Matsumoto T, Yamada H.1991.Effects of a polysaccharide fraction from the roots of Bupleurum falcatum L.on experimental gastric ulcer models in rats and mice.Journal of Pharmacy and Pharmacology, 43, 699-704.

    Wang B J, Liu C T, Tseng C Y, Yu Z R.2005.Antioxidant activity of Bupleurum kaoi Liu (Chao et Chuang) fractions fractionated by supercritical CO2.LWT-Food Science and Technology, 38, 281-287.

    Wang J P, Bu D P, Wang J Q, Huo X K, Guo T J, Wei H Y,Zhou LY, Rastani R R, Baumgard L H, Li F D.2010.Effect of saturated fatty acid supplemen-tation on production and metabolism indices in heat-stressed mid-lactation dairy cows.Journal of Dairy Science, 93, 4121-4127.

    Wheelock J B, Rhoads R P, VanBaale M J, Sanders S R,Baumgard L H.2010.Effects of heat stress on energetic metabolism in lactating Holstein cows.Journal of Dairy Science, 93, 644-655.

    Yamaguchi N, Kohno H, Tawara M, Odashima S.1985.Effect of saikosaponin derivatives upon the immune response against T-dependent and T-independent antigens in mice.International Journal of Immunopharmacology, 7, 827-832.

    Yen M H, Lin C C, Yen C M.1995.The immunomodulatory effect of saikosaponin derivatives and the root extract of Bupleurumkaoi in mice.Phytotherapy Research, 9, 351-358.

    Zhao W, Li J J, Yue S Q, Zhang L Y, Dou K F.2012.Antioxidant activity and hepatoprotective effect of a polysaccharide from Bei Chaihu (Bupleurum chinense DC).Carbohydrate Polymers, 89, 448-452.

    Zimbelman R B, Collier R J, Bilby T R.2013.Effects of utilizing rumen protected niacin on core body temperature as well as milk production and composition in lactating dairy cows during heat stress.Animal Feed Science and Technology,180, 26-33.

    国产人妻一区二区三区在| 天天躁日日操中文字幕| 十八禁人妻一区二区| 女人被狂操c到高潮| 可以在线观看毛片的网站| 婷婷亚洲欧美| 亚洲欧美日韩无卡精品| 国产精品女同一区二区软件 | 日本撒尿小便嘘嘘汇集6| 国产av一区在线观看免费| 国内精品一区二区在线观看| 波多野结衣高清无吗| 亚洲熟妇熟女久久| 久久九九热精品免费| 亚洲人成网站在线播| 搡老熟女国产l中国老女人| 国产乱人视频| 色综合站精品国产| 99久久精品国产亚洲精品| 99热这里只有是精品50| 1000部很黄的大片| 免费高清视频大片| 精品熟女少妇八av免费久了| 日韩大尺度精品在线看网址| 日韩人妻高清精品专区| 亚洲最大成人av| 精品熟女少妇八av免费久了| 国产在线男女| 精品人妻1区二区| 国产精品日韩av在线免费观看| 好男人在线观看高清免费视频| 亚洲在线自拍视频| 国产高潮美女av| 亚洲精品色激情综合| 性色avwww在线观看| 久久精品国产亚洲av天美| 国内精品一区二区在线观看| 亚洲综合色惰| 99riav亚洲国产免费| 亚洲不卡免费看| 成人国产综合亚洲| www.色视频.com| 啪啪无遮挡十八禁网站| 中文在线观看免费www的网站| 国产伦在线观看视频一区| 婷婷精品国产亚洲av在线| 国产三级黄色录像| 首页视频小说图片口味搜索| 韩国av一区二区三区四区| 婷婷精品国产亚洲av在线| or卡值多少钱| 久久久久性生活片| 一个人看视频在线观看www免费| 亚洲精品一区av在线观看| a级毛片a级免费在线| 别揉我奶头~嗯~啊~动态视频| 国内精品久久久久精免费| 在线天堂最新版资源| 日韩中字成人| 精品久久久久久久久av| 日韩精品中文字幕看吧| 一级av片app| 色综合欧美亚洲国产小说| 老女人水多毛片| 在线播放无遮挡| 亚洲午夜理论影院| 日韩欧美免费精品| 天美传媒精品一区二区| www.www免费av| 免费人成在线观看视频色| 中文字幕精品亚洲无线码一区| 成人av在线播放网站| 极品教师在线视频| 精品99又大又爽又粗少妇毛片 | 国产黄a三级三级三级人| 国产成人福利小说| 哪里可以看免费的av片| 亚洲最大成人av| 一区二区三区四区激情视频 | 亚洲久久久久久中文字幕| 怎么达到女性高潮| 在线观看免费视频日本深夜| 少妇的逼好多水| 简卡轻食公司| 久久久久久久久久成人| 国产黄色小视频在线观看| bbb黄色大片| 丁香六月欧美| 亚洲精品一区av在线观看| 日韩欧美免费精品| 丰满人妻熟妇乱又伦精品不卡| 波野结衣二区三区在线| 男女床上黄色一级片免费看| 欧美xxxx性猛交bbbb| 麻豆一二三区av精品| 此物有八面人人有两片| 精品免费久久久久久久清纯| 精品99又大又爽又粗少妇毛片 | 久久亚洲精品不卡| 毛片女人毛片| 9191精品国产免费久久| 一区福利在线观看| 欧美一区二区亚洲| 亚洲久久久久久中文字幕| 日本成人三级电影网站| 88av欧美| 国产亚洲欧美98| 天天一区二区日本电影三级| 日韩欧美一区二区三区在线观看| 1024手机看黄色片| 日本熟妇午夜| 亚洲美女视频黄频| www.www免费av| 免费观看精品视频网站| 日韩精品青青久久久久久| 俄罗斯特黄特色一大片| 精品久久久久久成人av| 亚洲精品在线美女| 99国产综合亚洲精品| 老司机午夜福利在线观看视频| 黄色一级大片看看| 黄片小视频在线播放| 久久久国产成人免费| 少妇的逼水好多| 欧美日本亚洲视频在线播放| 国产精品久久视频播放| 真人做人爱边吃奶动态| 少妇人妻一区二区三区视频| 成年人黄色毛片网站| 美女被艹到高潮喷水动态| 波多野结衣高清无吗| а√天堂www在线а√下载| 欧洲精品卡2卡3卡4卡5卡区| 91字幕亚洲| 国产精品精品国产色婷婷| 如何舔出高潮| 中文亚洲av片在线观看爽| 国产欧美日韩精品亚洲av| 美女 人体艺术 gogo| 欧美+日韩+精品| 午夜a级毛片| 亚洲欧美清纯卡通| 日韩欧美在线乱码| 久久精品国产亚洲av香蕉五月| 亚洲av免费高清在线观看| 欧美高清成人免费视频www| 精品久久久久久久久亚洲 | 亚洲天堂国产精品一区在线| 久久久久久九九精品二区国产| 俄罗斯特黄特色一大片| 男女那种视频在线观看| 亚洲美女黄片视频| 中亚洲国语对白在线视频| 日本一本二区三区精品| 美女大奶头视频| 最后的刺客免费高清国语| 中文字幕久久专区| 久久精品国产自在天天线| 高清毛片免费观看视频网站| 好看av亚洲va欧美ⅴa在| 国产精品av视频在线免费观看| 嫁个100分男人电影在线观看| 脱女人内裤的视频| 男人舔女人下体高潮全视频| 在现免费观看毛片| 欧美绝顶高潮抽搐喷水| 国产精品一区二区三区四区久久| 久久久久性生活片| 国产精品久久久久久久电影| 露出奶头的视频| 亚洲欧美精品综合久久99| 免费看光身美女| 亚洲,欧美精品.| 午夜激情欧美在线| 欧美日韩综合久久久久久 | 亚洲av第一区精品v没综合| 国产aⅴ精品一区二区三区波| 国产精品综合久久久久久久免费| 免费人成视频x8x8入口观看| 午夜视频国产福利| 丰满的人妻完整版| 亚洲成a人片在线一区二区| 久久久久国产精品人妻aⅴ院| 亚洲国产精品sss在线观看| 国产高潮美女av| 欧美日韩国产亚洲二区| 国产精品亚洲一级av第二区| 自拍偷自拍亚洲精品老妇| 久久婷婷人人爽人人干人人爱| 我要看日韩黄色一级片| 简卡轻食公司| 欧美日韩亚洲国产一区二区在线观看| 免费观看精品视频网站| 亚洲 欧美 日韩 在线 免费| 自拍偷自拍亚洲精品老妇| 亚洲av美国av| 99热这里只有是精品50| 精品一区二区三区人妻视频| 丰满人妻一区二区三区视频av| 欧美黑人巨大hd| 亚洲男人的天堂狠狠| 久久精品国产亚洲av涩爱 | 久久久成人免费电影| 99国产精品一区二区蜜桃av| 免费看日本二区| 国产午夜精品论理片| 国产精品久久久久久亚洲av鲁大| 精品久久久久久久久av| 国产精品嫩草影院av在线观看 | 亚洲第一电影网av| 亚洲一区二区三区色噜噜| 久久久久久大精品| 老司机福利观看| 成人永久免费在线观看视频| 亚洲精品亚洲一区二区| 99在线人妻在线中文字幕| 欧美日韩福利视频一区二区| 国产精品电影一区二区三区| 2021天堂中文幕一二区在线观| 欧美在线一区亚洲| 久久草成人影院| 看十八女毛片水多多多| 免费高清视频大片| 又爽又黄无遮挡网站| av福利片在线观看| 国产av不卡久久| 成年免费大片在线观看| av天堂中文字幕网| 亚洲内射少妇av| 亚洲成人久久爱视频| 成人一区二区视频在线观看| 国产高清视频在线播放一区| 亚洲av一区综合| 直男gayav资源| 大型黄色视频在线免费观看| 91在线精品国自产拍蜜月| 久久人妻av系列| 男人舔奶头视频| 听说在线观看完整版免费高清| 亚洲男人的天堂狠狠| 91久久精品电影网| 国产黄色小视频在线观看| 久久人妻av系列| 亚洲美女黄片视频| 午夜免费激情av| 淫妇啪啪啪对白视频| 无遮挡黄片免费观看| 嫁个100分男人电影在线观看| 看黄色毛片网站| 男人舔女人下体高潮全视频| 国产真实乱freesex| 成人永久免费在线观看视频| 少妇丰满av| 午夜福利在线观看吧| 99久久成人亚洲精品观看| 国产极品精品免费视频能看的| 麻豆国产97在线/欧美| 日韩亚洲欧美综合| 成人一区二区视频在线观看| 88av欧美| 国产aⅴ精品一区二区三区波| 亚洲国产精品999在线| 精品人妻熟女av久视频| 亚洲一区二区三区色噜噜| 一个人观看的视频www高清免费观看| 中文字幕高清在线视频| 亚洲精品粉嫩美女一区| 久久国产精品人妻蜜桃| 黄色女人牲交| 亚洲不卡免费看| 亚洲美女黄片视频| 非洲黑人性xxxx精品又粗又长| 欧美三级亚洲精品| 在线观看午夜福利视频| 伊人久久精品亚洲午夜| 欧美一区二区亚洲| 国产亚洲精品av在线| 国产精品一区二区三区四区久久| 国产高清三级在线| 国产精品久久久久久精品电影| 夜夜夜夜夜久久久久| 色哟哟哟哟哟哟| 欧美成人a在线观看| 久久久久久久久久成人| 免费大片18禁| 久久久久久久久中文| 亚洲国产欧洲综合997久久,| 国产成人a区在线观看| 国产一区二区在线观看日韩| 禁无遮挡网站| 老师上课跳d突然被开到最大视频 久久午夜综合久久蜜桃 | 免费观看的影片在线观看| 极品教师在线视频| 两性午夜刺激爽爽歪歪视频在线观看| 村上凉子中文字幕在线| 99国产综合亚洲精品| 如何舔出高潮| 在线观看66精品国产| 美女黄网站色视频| 国产单亲对白刺激| 亚洲第一区二区三区不卡| 日本精品一区二区三区蜜桃| 久久久久性生活片| 亚洲国产高清在线一区二区三| 哪里可以看免费的av片| 熟女人妻精品中文字幕| 亚洲国产日韩欧美精品在线观看| av欧美777| 美女xxoo啪啪120秒动态图 | 1000部很黄的大片| 床上黄色一级片| 97超级碰碰碰精品色视频在线观看| 亚洲色图av天堂| 亚洲自偷自拍三级| 99riav亚洲国产免费| 脱女人内裤的视频| 久久人人精品亚洲av| 成人午夜高清在线视频| 日韩欧美在线二视频| 国产极品精品免费视频能看的| 亚洲18禁久久av| 国产日本99.免费观看| 色5月婷婷丁香| 午夜免费成人在线视频| 天美传媒精品一区二区| 欧美成人免费av一区二区三区| 亚洲中文字幕日韩| 免费看美女性在线毛片视频| 亚洲avbb在线观看| 亚洲 欧美 日韩 在线 免费| 一个人免费在线观看的高清视频| 亚洲精品乱码久久久v下载方式| 麻豆久久精品国产亚洲av| 极品教师在线免费播放| av黄色大香蕉| 免费看a级黄色片| 国产欧美日韩一区二区精品| 国产av麻豆久久久久久久| 午夜福利欧美成人| 欧美色视频一区免费| 免费无遮挡裸体视频| 日韩欧美免费精品| 在线十欧美十亚洲十日本专区| 欧美日韩乱码在线| 99国产精品一区二区三区| av在线天堂中文字幕| 国产在线男女| 亚洲av电影在线进入| 免费在线观看日本一区| 国产精品一区二区三区四区久久| 成人美女网站在线观看视频| 国产精品,欧美在线| 欧美国产日韩亚洲一区| 丰满的人妻完整版| 又黄又爽又刺激的免费视频.| 日韩欧美一区二区三区在线观看| 国产aⅴ精品一区二区三区波| 一进一出好大好爽视频| 亚洲人成网站高清观看| 少妇的逼好多水| 成人一区二区视频在线观看| 欧美一区二区国产精品久久精品| 在线播放无遮挡| 九九热线精品视视频播放| 国产伦精品一区二区三区四那| 国产成年人精品一区二区| 国产在线精品亚洲第一网站| 精品午夜福利在线看| 真实男女啪啪啪动态图| 国产av不卡久久| 国产精品电影一区二区三区| 淫秽高清视频在线观看| 日日摸夜夜添夜夜添av毛片 | 三级毛片av免费| 日韩免费av在线播放| 一级作爱视频免费观看| 91午夜精品亚洲一区二区三区 | 人人妻人人看人人澡| 国产一区二区亚洲精品在线观看| 国产亚洲精品av在线| 18禁在线播放成人免费| 欧美绝顶高潮抽搐喷水| 啪啪无遮挡十八禁网站| 精品不卡国产一区二区三区| 欧美成狂野欧美在线观看| 999久久久精品免费观看国产| 午夜精品在线福利| 国产乱人视频| 国产精品亚洲av一区麻豆| 成人三级黄色视频| 色av中文字幕| www.色视频.com| 国产又黄又爽又无遮挡在线| 五月玫瑰六月丁香| 中文字幕高清在线视频| 制服丝袜大香蕉在线| 国产探花极品一区二区| 亚洲精华国产精华精| 九九在线视频观看精品| 亚洲av免费高清在线观看| 悠悠久久av| 欧美xxxx性猛交bbbb| av在线观看视频网站免费| 国产成人欧美在线观看| 久久婷婷人人爽人人干人人爱| 中文字幕人妻熟人妻熟丝袜美| 欧美黄色片欧美黄色片| 天堂av国产一区二区熟女人妻| 女生性感内裤真人,穿戴方法视频| 小蜜桃在线观看免费完整版高清| 日本免费a在线| 亚洲最大成人中文| 久久久久国内视频| 12—13女人毛片做爰片一| 精品一区二区三区视频在线| 日韩欧美国产一区二区入口| 国产v大片淫在线免费观看| 一区福利在线观看| 久久久久久久亚洲中文字幕 | 国产成+人综合+亚洲专区| 狠狠狠狠99中文字幕| 亚洲av二区三区四区| 一本久久中文字幕| 一进一出抽搐gif免费好疼| 国产美女午夜福利| 精品人妻视频免费看| 亚洲久久久久久中文字幕| 欧美午夜高清在线| 国产色婷婷99| 久久久精品欧美日韩精品| 欧美绝顶高潮抽搐喷水| 一边摸一边抽搐一进一小说| 国产高清视频在线播放一区| 最新在线观看一区二区三区| 久久精品久久久久久噜噜老黄 | 国产精品久久久久久久电影| 国产精品美女特级片免费视频播放器| 色在线成人网| 高清在线国产一区| 国产av不卡久久| 9191精品国产免费久久| 69av精品久久久久久| 久久香蕉精品热| 麻豆成人午夜福利视频| 亚洲精品一区av在线观看| 精品日产1卡2卡| 亚洲激情在线av| 国产探花在线观看一区二区| 听说在线观看完整版免费高清| 亚洲国产精品合色在线| 国产亚洲av嫩草精品影院| 亚洲成人久久性| 国产人妻一区二区三区在| 午夜影院日韩av| 欧洲精品卡2卡3卡4卡5卡区| 国产欧美日韩精品亚洲av| 亚洲午夜理论影院| 毛片一级片免费看久久久久 | 成人美女网站在线观看视频| 久久精品久久久久久噜噜老黄 | 亚洲第一区二区三区不卡| 可以在线观看的亚洲视频| 2021天堂中文幕一二区在线观| 一卡2卡三卡四卡精品乱码亚洲| 国产精品一及| 久久久久久久久中文| 亚洲七黄色美女视频| 日本成人三级电影网站| 日韩 亚洲 欧美在线| 久久国产乱子伦精品免费另类| 久久伊人香网站| 一二三四社区在线视频社区8| 在线国产一区二区在线| 一个人观看的视频www高清免费观看| 亚洲 欧美 日韩 在线 免费| 在线看三级毛片| 亚洲片人在线观看| 99热这里只有是精品50| 18禁在线播放成人免费| 小说图片视频综合网站| 午夜激情欧美在线| 久久精品夜夜夜夜夜久久蜜豆| 久久国产精品人妻蜜桃| 听说在线观看完整版免费高清| 国产乱人伦免费视频| 欧美色视频一区免费| 午夜影院日韩av| 国产av一区在线观看免费| 有码 亚洲区| 熟妇人妻久久中文字幕3abv| 嫁个100分男人电影在线观看| 嫩草影视91久久| 精品久久久久久成人av| 九色成人免费人妻av| 一个人看视频在线观看www免费| 性色avwww在线观看| 欧美+亚洲+日韩+国产| 欧美另类亚洲清纯唯美| 国产成人福利小说| 亚洲va日本ⅴa欧美va伊人久久| 性色avwww在线观看| 久久精品人妻少妇| 日韩欧美在线二视频| 精品久久久久久久久av| 黄色一级大片看看| 好男人在线观看高清免费视频| 村上凉子中文字幕在线| 久久国产精品人妻蜜桃| 偷拍熟女少妇极品色| 欧美区成人在线视频| 非洲黑人性xxxx精品又粗又长| 国内揄拍国产精品人妻在线| 丁香六月欧美| 日日干狠狠操夜夜爽| 国内精品一区二区在线观看| 一二三四社区在线视频社区8| 最近视频中文字幕2019在线8| 十八禁国产超污无遮挡网站| 久久久久久久久大av| 高清在线国产一区| 男女做爰动态图高潮gif福利片| 亚洲久久久久久中文字幕| 少妇被粗大猛烈的视频| x7x7x7水蜜桃| 亚洲美女黄片视频| 一级av片app| 久久久精品大字幕| 麻豆国产97在线/欧美| 欧美日韩国产亚洲二区| 女人十人毛片免费观看3o分钟| 中文亚洲av片在线观看爽| 精品人妻熟女av久视频| 村上凉子中文字幕在线| 欧美中文日本在线观看视频| 久久国产乱子免费精品| 欧美另类亚洲清纯唯美| avwww免费| 中文字幕人妻熟人妻熟丝袜美| 久久久久久久久久黄片| 精品一区二区三区av网在线观看| 变态另类丝袜制服| 亚洲中文日韩欧美视频| 深爱激情五月婷婷| 一本综合久久免费| 日日干狠狠操夜夜爽| .国产精品久久| 青草久久国产| 精品一区二区三区视频在线观看免费| 99精品久久久久人妻精品| 色噜噜av男人的天堂激情| 日日摸夜夜添夜夜添小说| 日日干狠狠操夜夜爽| 欧美一区二区精品小视频在线| 日韩欧美在线二视频| 淫秽高清视频在线观看| 亚洲成人免费电影在线观看| 国产单亲对白刺激| 亚洲内射少妇av| 欧美不卡视频在线免费观看| 欧美日韩福利视频一区二区| 黄色女人牲交| 色5月婷婷丁香| 男人和女人高潮做爰伦理| 少妇被粗大猛烈的视频| 成人精品一区二区免费| 国产精品久久视频播放| 午夜影院日韩av| 人妻丰满熟妇av一区二区三区| 69av精品久久久久久| 欧美+亚洲+日韩+国产| 中文在线观看免费www的网站| 少妇裸体淫交视频免费看高清| 999久久久精品免费观看国产| 人妻久久中文字幕网| 欧美日韩中文字幕国产精品一区二区三区| 波野结衣二区三区在线| 最新在线观看一区二区三区| 亚洲熟妇中文字幕五十中出| 国产探花极品一区二区| 日韩有码中文字幕| 九九久久精品国产亚洲av麻豆| 久久久久免费精品人妻一区二区| 一本久久中文字幕| 精华霜和精华液先用哪个| 看免费av毛片| 老鸭窝网址在线观看| 亚洲精品一区av在线观看| 高潮久久久久久久久久久不卡| 免费搜索国产男女视频| 国产男靠女视频免费网站| 午夜精品一区二区三区免费看| 日本a在线网址| 日韩欧美 国产精品| 亚洲久久久久久中文字幕| 亚洲性夜色夜夜综合| 麻豆久久精品国产亚洲av| 亚洲不卡免费看| 免费大片18禁| 国内少妇人妻偷人精品xxx网站| 级片在线观看| 两个人视频免费观看高清| 日韩精品青青久久久久久| 久久久久久久亚洲中文字幕 | 欧美精品啪啪一区二区三区| 热99re8久久精品国产| 99热只有精品国产| 欧美不卡视频在线免费观看| 97超级碰碰碰精品色视频在线观看| 波多野结衣高清作品| 白带黄色成豆腐渣| 欧美色欧美亚洲另类二区| 久久人人爽人人爽人人片va | 狠狠狠狠99中文字幕| 此物有八面人人有两片| 久久久精品大字幕|