芹菜素亚慢性暴露对小鼠、大鼠肝脏功能及脂质过氧化的影响

发布时间：2018-06-13 19:43

本文选题：芹菜素 + 雄性大鼠　；参考：《兰州大学》2014年硕士论文

【摘要】：目的通过芹菜素灌胃,观察芹菜素在不同剂量、不同时间对雄性昆明种小鼠和SD大鼠的血液生化指标、肝脏氧化还原指标和血液氧化还原指标的影响,探讨其剂量效应,为芹菜素的深入研究和进一步的开发利用提供科学依据。方法1.将200只健康SPF级雄性昆明种小鼠按体重随机分为4组,分别为阴性对照组(生理盐水10ml/kg)、低剂量组(AP252mg/kg)、中剂量组(AP504mg/kg)和高剂量组(AP1008mg/kg),每组50只。每天灌胃1次,每周6次,连续灌胃5周。在实验第7天、14天、21天、28天、35天,分批眼眶内眦静脉丛取血,检测血清总蛋白(TP)、白蛋白(ALB)、球蛋白(GLO)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、谷氨酰转肽酶(GGT)和碱性磷酸酶(ALP)活力。颈椎脱臼处死小鼠后取肝脏,制备10%肝脏组织匀浆。比色法检测肝脏组织匀浆中总抗氧化能力(T-AOC)、还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)吸光值,并计算各指标酶活力。 2.将48只健康SPF级雄性SD大鼠按体重随机分为4组,分别为对照组(生理盐水10ml/kg)、低剂量组(AP234mg/kg)、中剂量组(AP468mg/kg)和高剂量组(AP936mg/kg),每组12只。每天灌胃1次,每周6次,连续灌胃5周。实验第35天心脏穿刺取血5ml,检测血清总蛋白(TP)、白蛋白(ALB)、球蛋白(GLO)、丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)活力和血清丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、总抗氧化能力(T-AOC)、谷胱甘肽过氧化物酶(GSH-Px)、谷胱甘肽(GSH)含量。处死大鼠,取出肝脏后-20℃保存待用。检测前制备10%肝脏组织匀浆,检测丙二醛(MDA)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、总抗氧化能力(T-AOC)、谷胱甘肽过氧化物酶(GSH-Px)、谷胱甘肽(GSH)含量,并计算各指标酶活力。在光镜下观察大鼠肝脏组织的病理学改变。结果1.体重小鼠体重14天中剂量组(31.27±3.32g)与对照组(35.53±2.84g)比较降低,差异有统计学意义(P0.05)；大鼠体重呈现增长趋势,各剂量组与对照组比较差异均无统计学意义(P0.05)。 2.脏器系数小鼠肝脏系数14天中剂量组(3.96±0.44)×10-2与对照组(4.42±0.63)×10-2比较降低,差异有统计学意义(P0.05)；28天低剂量组(4.23±0.39)×10-2、中剂量组(3.88±0.40)×10-2、高剂量组(4.15±0.41)×10-2与对照组(4.57±0.38)×10-2比较降低,差异有统计学意义(P0.05)；35天高剂量组(4.15±0.39)×10-2与对照组(4.47±0.28)X10-2比较降低,差异有统计学意义(P0.05)。大鼠肝脏重量、脾脏重量,肝/体比(%)、脾/体比(‰),与各自对照组比较差异均无统计学意义(P0.05)。 3.血清生化小鼠7天高剂量组血清TP、ALB、ALT、AST、ALP与相应对照组比较升高,差异有统计学意义(P0.05)14天中剂量组血清中TP、 ALB、ALT、AST与相应对照组比较升高,差异有统计学意义(P0.05)；21天低剂量组血清中TP、ALB、ALT、AST与相应对照组比较升高,差异有统计学意义(P0.05)；35天ALB、ALT、AST和ALP低剂量组、中剂量组、高剂量组与对照组比较降低,差异有统计学意义(P0.05)。大鼠血清TP、ALB、 GLO低、中、高剂量组与对照组比较降低,差异有统计学意义(P0.05)。ALT、 AST低、中、高剂量组与各自对照组比较差异均无统计学意义(P0.05)。 4.大鼠血清氧化还原酶大鼠血清SOD低剂量组(178.43±7.22U/mg)与对照组(159.07±22.07U/mg)比较升高,差异有统计学意义(P0.05)；CAT高剂量组(9.45±3.90U/m1)与对照组(5.31±1.79U/m1)比较升高,差异有统计学意义(P0.05); GSH-Px低剂量组(1922.09±180.38U/mg)与对照组(2154.22±173.41U/mg)比较降低,差异有统计学意义(P0.05)；T-AOC低(9.00±3.08U/mg)、中(7.85±2.40U/mg)、高剂量组(9.59±2.32U/mg)与对照组(12.03±2.82U/mg)比较降低,差异有统计学意义(P0.05)。 5.小鼠肝脏氧化还原酶小鼠肝脏T-AOC在7天(1.68±0.82U/mmg)、14天(1.04±0.42U/mmg)、28天(1.08±0.45U/mg)高剂量组与对照组比较均降低,且差异有统计学意义(P0.05)；21天中剂量组(2.04±1.05)与对照组(1.43±1.58)比较升高,差异有统计学意义(P0.05); GSH/GSSG在28天低剂量组(1.70±1.61)与对照组(1.70±1.61)比较升高,差异有统计学意义(P0.05)；MDA在21天高剂量组(1.07±0.73nmol/mg)与对照组(0.35±0.21nmol/mg)比较升高,差异有统计学意义(P0.05)；SOD在28天高剂量组(100.73±10.87U/mg)与对照组(111.60±3.36U/mg)比较降低,差异有统计学意义(P0.05)；35天中剂量组(91.30±13.99U/mg)与对照组(108.00±6.04U/mg)比较降低,差异有统计学意义(P0.05); CAT在7天高剂量组(50.60±28.06U/m1)与对照组(22.70±11.15U/m1)比较升高,差异有统计学意义(P0.05)；35天高剂量组(115.19±36.43U/ml)与对照组(69.84±32.10U/ml)比较升高,差异有统计学意义(P0.05):GSH-Px在21天低剂量组(70.21±40.61U/mg)与对照组(40.07±9.18U/mg)比较升高,差异有统计学意义(P0.05)；28天高剂量组(33.43±17.77U/mg)与对照组(50.85±13.12U/mg)比较降低,差异有统计学意义(P0.05)。 6.大鼠肝脏氧化还原酶大鼠肝脏MDA低(0.66±0.13nmol/mg).中(0.31±0.05nmol/mg).高(0.40±0.07nmol/mg)剂量组与对照组(0.48±0.06nmol/mg)比较差异有统计学意义(P0.05)；NOS低剂量组(0.74±0.19U/ml)与对照组(0.49±0.17U/ml)比较升高,差异有统计学意义(P0.05)；SOD中(19.59±3.16U/mg)、高(19.48±1.91U/mg)剂量组与对照组(24.81±2.72U/mg)比较均降低,差异有统计学意义(P0.05);CAT低(53.37±8.89U/ml)、中(36.48±3.79U/ml)、高(42.97±6.11U/ml)剂量组与对照组(68.66±6.50U/ml)比较降低,差异有统计学意义(P0.05);T-AOC低(0.44±0.02U/mg)、中(0.44±0.09U/mg)、高(0.37±0.08U/mg)剂量组与对照组(0.74±0.12U/mg)比较降低,差异有统计学意义(P0.05)；GSH-Px低(290.37±57.71U/mg)、中(346.79±41.75U/mg)高(243.18±42.89U/mg)剂量组与对照组(403.66±92.83U/mg)比较降低,差异有统计学意义(P0.05)。结论芹菜素亚慢性暴露可影响雄性小鼠和大鼠的肝脏功能；芹菜素在不同剂量、不同时间对不同品系动物在不同组织表现出的抗/促氧化作用并不一致,提示黄酮类化合物在不同剂量下表现出的作用是不同的,作为膳食抗氧化剂应用时,应该有一个适宜的摄入时间和剂量。
[Abstract]:Objective To observe the effect of apigenin on the blood biochemical index, liver redox index and blood oxidation-reduction index of male Kunming mice and SD rats in different doses and time, and to provide a scientific basis for the in-depth study and further development of apigenin in different doses and time.
Method 1. 200 healthy SPF male Kunming mice were randomly divided into 4 groups according to their weight. They were negative control group (10ml/kg), low dose group (AP252mg/kg), medium dose group (AP504mg/kg) and high dose group (AP1008mg/kg), 50 rats in each group, 1 times a day, 6 times a week for 5 weeks. The experiment was seventh days, 21 days, 28 days, 35 days, Serum total protein (TP), albumin (ALB), globulin (GLO), alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutamine transaminopeptidase (GGT) and alkaline phosphatase (ALP) activity were detected in the orbital canthus plexus, and 10% liver tissue homogenate was prepared after the dehydration of the cervical vertebra in mice. The liver tissue was detected by colorimetric assay. The total antioxidant capacity (T-AOC) in the homogenate, reduced glutathione / oxidized glutathione (GSH/GSSG), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and calculated the activity of each index.
2. 48 healthy SPF grade male SD rats were randomly divided into 4 groups according to their weight. They were the control group (physiological saline 10ml/kg), low dose group (AP234mg/kg), middle dose group (AP468mg/kg) and high dose group (AP936mg/kg), each group was filled with 1 times a day, 6 times a week for 5 weeks, and thirty-fifth days of heart puncture, blood 5ml, and serum total protein (TP) were detected. Albumin (ALB), globulin (GLO), alanine aminotransferase (ALT), aspartate aminotransferase (AST) activity and serum malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), glutathione (GSH) content. 10% liver tissue homogenate was prepared before detection, and the content of malondialdehyde (MDA), nitric oxide synthase (NOS), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (T-AOC), glutathione peroxidase (GSH-Px), glutathione (GSH), and the enzyme activity were calculated. The pathological changes of liver tissues of rats were observed under light microscope. Change.
Results the weight of 1. weight mice in 14 days (31.27 + 3.32g) was lower than that of the control group (35.53 + 2.84g). The difference was statistically significant (P0.05), and the weight of the rats showed a trend of growth, and there was no significant difference between the different dose groups and the control group (P0.05).
The liver coefficient of the 2. organ coefficient mice was 14 days (3.96 + 0.44) x 10-2 and the control group (4.42 + 0.63) x 10-2. The difference was statistically significant (P0.05); the low dose group for 28 days (4.23 + 0.39) x 10-2, the middle dose group (3.88 +) * * *, the high dose group was compared with the control group. The difference was statistically significant. The study significance (P0.05); the 35 day high dose group (4.15 + 0.39) x 10-2 and the control group (4.47 + 0.28) X10-2 decreased, the difference was statistically significant (P0.05). The liver weight, spleen weight, liver / body ratio (%), spleen / body ratio (%), and the comparison of the control group were not statistically significant (P0.05).
3. the serum TP, ALB, ALT, AST, ALP in the high dose group of serum biochemical mice were higher than those in the corresponding control group (P0.05), the difference was statistically significant (P0.05) in the serum of 14 days, TP, ALB, ALT, AST were higher than the corresponding control group, and the difference was statistically significant (P0.05), and the low dose group was higher than the corresponding control group at the low dose of 21 days. The difference was statistically significant (P0.05), 35 days ALB, ALT, AST and ALP low dose group, middle dose group, the high dose group was lower than the control group, the difference was statistically significant (P0.05). The serum TP, ALB, GLO low, the high dose group was lower than the control group, the difference was statistically significant (P0.05).ALT, low AST, high dose group and their control groups The difference was not statistically significant (P0.05).
The serum SOD low dose group (178.43 + 7.22U/mg) and the control group (159.07 + 22.07U/mg) were higher in the 4. rats. The difference was statistically significant (P0.05); the high dose group of CAT (9.45 + 3.90U/m1) was higher than the control group (5.31 + 1.79U/m1), and the difference was statistically significant (P0.05); the low dose group of GSH-Px (1922.09 + 180.38U/mg) was compared with the control group (5.31 + 1.79U/m1). The control group (2154.22 + 173.41U/mg) was lower, the difference was statistically significant (P0.05); T-AOC was low (9 + 3.08U/mg), middle (7.85 + 2.40U/mg), high dose group (9.59 + 2.32U/mg) and control group (12.03 + 2.82U/mg) decreased, the difference was statistically significant (P0.05).
5. the liver T-AOC of mice liver oxidoreductase was 7 days (1.68 + 0.82U/mmg), 14 days (1.04 + 0.42U/mmg), 28 days (1.08 + 0.45U/mg) in high dose group decreased with the control group, and the difference was statistically significant (P0.05). In 21 days, the dose group (2.04 + 1.05) was significantly higher than that in the group (1.43 + 1.58), and the difference was statistically significant (P0.05); GSH/GSSG In the 28 day low dose group (1.70 + 1.61) and the control group (1.70 + 1.61), the difference was statistically significant (P0.05); MDA was higher in the 21 day high dose group (1.07 + 0.73nmol/mg) and the control group (0.35 + 0.21nmol/mg), and the difference was statistically significant (P0.05); SOD in the high dose group 28 days (100.73 + 10.87U/mg) and the control group (111.60 + 3.36U/mg) ratio. The difference was statistically significant (P0.05); in the 35 day medium dose group (91.30 + 13.99U/mg) and the control group (108 + 6.04U/mg), the difference was statistically significant (P0.05); CAT was higher in the 7 day high dose group (50.60 + 28.06U/m1) and the control group (22.70 + 11.15U/m1), the difference was statistically significant (P0.05), and the 35 day high dose group (115.19 + 3). 6.43U/ml) compared with the control group (69.84 + 32.10U/ml), the difference was statistically significant (P0.05): GSH-Px was higher in the 21 day low dose group (70.21 + 40.61U/mg) and the control group (40.07 + 9.18U/mg), and the difference was statistically significant (P0.05); the 28 day high dose group (33.43 + 17.77U/mg) was lower than the control group (50.85 + 13.12U/mg), and the difference was statistically significant. Learning meaning (P0.05).
The liver MDA of the 6. rat liver oxidoreductase was low (0.66 + 0.13nmol/mg). (0.31 + 0.13nmol/mg). The high (0.40 + 0.07nmol/mg) dose group was significantly different from the control group (0.48 + 0.06nmol/mg), and the NOS low dose group (0.74 + 0.19U/ml) was higher than that of the group (0.49 + 0.17U/ml), and the difference was statistically significant (P0.05); S OD (19.59 + 3.16U/mg), high (19.48 + 1.91U/mg) dose group and control group (24.81 + 2.72U/mg) were lower, the difference was statistically significant (P0.05), CAT low (53.37 + 8.89U/ml), (36.48 + 3.79U/ml), high (42.97 + 6.11U/ml) dose group and the control group (68.66 + 6.50U/ml) decreased, the difference was statistically significant (P0.05); low T-AOC (0.44 + 42.97). G), in the medium (0.44 + 0.09U/mg), the high (0.37 + 0.08U/mg) dose group was lower than the control group (0.74 + 0.12U/mg), the difference was statistically significant (P0.05), the GSH-Px low (290.37 + 57.71U/mg), the middle (346.79 + 41.75U/mg) high (243.18 + 42.89U/mg) dose group were lower than the control group (403.66 + 92.83U/mg), and the difference was statistically significant (P0.05).
Conclusion apigenin subchronic exposure can affect the liver function of male and female mice. Apigenin has different anti / anti oxidative effects on different tissues at different doses and different time, suggesting that the effects of flavonoids in different doses are different and are used as dietary antioxidants. There should be a suitable intake time and dose.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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