低剂量铜暴露对大鼠肝功能和炎性因子表达的影响及乳酸菌干预作用
发布时间：2018-12-25 21:15
【摘要】：目的:1.探讨低剂量的铜暴露对大鼠肝脏功能和肝脏组织炎性因子的影响;2.探讨乳酸菌(LAB)对铜暴露引起的大鼠肝脏功能和炎性细胞因子表达水平的干预效果。方法:大鼠适应性喂养7天后,按体重随机分为8组:高铜组(14mg/kg)、中铜组(3.5mg/kg组)、低铜组(0.875mg/kg组)、高铜+乳酸菌组(14mg/kg+乳酸菌组)、中铜+乳酸菌组(3.5mg/kg组+乳酸菌组)、低铜+乳酸菌组(0.875mg/kg组+乳酸菌组)、乳酸菌组和空白对照组,每组按相应剂量连续性灌胃硫酸铜和乳酸菌30天。1.实验结束后,大鼠腹主动脉处取血,处死大鼠后,取大鼠肝、脾和胸腺等脏器,观察大鼠肝、脾和胸腺指数;2.对肝脏组织进行石蜡包埋,切片并进行HE染色,显微镜观察肝脏的病理变化;3.检测血清中的与肝功能相关的蛋白:谷丙转氨酶(ALT)、谷草转氨酶(AST)和白蛋白(ALB)含量;4.ICP-MS(电感耦合等离子体质谱仪)法测定大鼠血清中的铜含量;5.检测肝脏组织中与氧化应激相关的分子:超氧化物歧化酶(SOD)、丙二醛(MDA)和谷胱甘肽过氧化物酶(GSH-Px)含量;6.RT-PCR法检测肝脏组织中白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-12(IL-12)、超氧化物歧化酶和肿瘤坏死因子αTNF-αm RNA的相对表达量。结果:1.各组间大鼠的肝脏、脾和胸腺指数未见明显差异(P0.05);2.通过观察肝组织病理切片,发现高铜组、中铜组和低铜组大鼠肝组织在铜处理下均有不同程度损伤的发生,其中高铜组最为明显;在加入乳酸菌干预后,高铜组大鼠的肝损伤得到一定程度的改善;3.与对照组相比,高铜组、中铜和低铜组的AST、ALT显著增高(P0.05);ALB降低明显(P0.05);经乳酸菌干预后,不同剂量铜暴露组中的ALB均显著升高(P0.05),而只有中铜组、低铜组AST和ALT有明显改善(P0.05)。4.与对照组相比,不同剂量铜暴露组中的血清铜水平均显著升高(P0.05);乳酸菌干预后,三组的血清铜水平均减少,但无统计学意义(P0.05)。5.与对照组相比,不同剂量铜暴露组中的MDA含量均显著升高(P0.05),只有高铜组和中铜组的SOD活性和SOD m RNA显著降低(P0.05);乳酸菌干预后,在高铜组和中铜组SOD和MDA的水平明显恢复(P0.05)。6.不同剂量的铜暴露均导致TNF-α、IL-1β、IL-6和IL-10 m RNA升高,IL-12 m RNA在高铜组中也升高并且有统计学意义(P0.05);乳酸菌干预后,TNF-α、IL-1β和IL-6 m RNA均明显降低(P0.05)。结论:1.不同剂量的铜处理均可引起AST、ALT和ALB的显著改变,表明低剂量铜暴露能够引起大鼠肝脏功能下降;2.乳酸菌干预后,AST、ALT和ALB均有不同程度的改善,提示乳酸菌对铜暴露引起的肝脏功能下降的保护作用;3.低铜组引起IL-1β、IL-6和IL-10 m RNA表达上升,提示低剂量铜暴露即可引起大鼠肝脏炎性损伤;4.乳酸菌干预后,IL-1β、IL-6和TNF-αm RNA均明显降低,提示乳酸菌对铜暴露引起的肝损伤的保护作用可能与炎性细胞因子的释放有关。
[Abstract]:Objective: 1. To investigate the effects of low dose copper exposure on liver function and hepatic inflammatory factors in rats. 2. To investigate the effect of lactic acid bacteria (LAB) on liver function and inflammatory cytokine expression induced by copper exposure in rats. Methods: after 7 days of adaptive feeding, rats were randomly divided into 8 groups according to their body weight: high copper group (14mg/kg), medium copper group (3.5mg/kg group), low copper group (0.875mg/kg group) and high copper lactobacillus group (14mg/kg lactobacillus group). Medium copper lactobacillus group (3.5mg/kg group), low copper lactobacillus group (0.875mg/kg group lactic acid bacteria group), lactic acid bacteria group and blank control group, each group in accordance with the corresponding dose of continuous gastric administration of copper sulfate and lactic acid bacteria for 30 days. 1. At the end of the experiment, blood was taken from the abdominal aorta of the rats. After the rats were killed, the liver, spleen and thymus were taken and the indexes of liver, spleen and thymus were observed. 2. The liver tissue was embedded in paraffin, sectioned and stained with HE, and the pathological changes of liver were observed by microscope. 3. Serum proteins related to liver function: glutamate-pyruvic transaminase (ALT), aspartate aminotransferase (AST) and albumin (ALB), serum copper (Cu) in rats by 4.ICP-MS (inductively coupled plasma mass spectrometer). The contents of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in liver tissues were determined. Interleukin-1 尾 (IL-1 尾), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12 (IL-12) in liver tissue were detected by 6.RT-PCR. The relative expression of superoxide dismutase and tumor necrosis factor 伪 TNF- 伪 m RNA. Results: 1. There was no significant difference in liver, spleen and thymus index among groups (P0.05). By observing the pathological sections of liver tissue, it was found that the liver tissues of high copper group, middle copper group and low copper group all had different degrees of damage under copper treatment, among which the high copper group was the most obvious. The liver injury in high copper group was improved to some extent after the intervention of lactobacillus. Compared with the control group, the AST,ALT of high copper group, medium copper group and low copper group was significantly higher (P0.05); ALB decreased significantly (P0.05); After the intervention of lactic acid bacteria, the ALB in copper exposure group increased significantly (P0.05), but only in medium copper group, AST and ALT were significantly improved in low copper group (P0.05). Compared with the control group, the levels of serum copper in different doses of copper exposure group were significantly higher (P0.05); after the intervention of lactic acid bacteria, the serum copper level of the three groups decreased, but there was no statistical significance (P0.05). Compared with the control group, the MDA content in the copper exposure group was significantly higher than that in the control group (P0.05), but the SOD activity and SOD m RNA decreased significantly in the high copper group and the middle copper group (P0.05). After the intervention of lactic acid bacteria, the levels of SOD and MDA in high copper group and middle copper group recovered significantly (P0.05). The levels of TNF- 伪, IL-1 尾, IL-6 and IL-10 m RNA were increased by different doses of copper exposure, and IL-12 m RNA was also increased in high copper group (P0.05). TNF- 伪, IL-1 尾 and IL-6 m RNA were significantly decreased after lactobacillus intervention (P0.05). Conclusion: 1. Different doses of copper could cause significant changes in AST,ALT and ALB, which indicated that low dose copper exposure could induce the decrease of liver function in rats. 2. After the intervention of lactic acid bacteria, both AST,ALT and ALB were improved to some extent, suggesting the protective effect of lactic acid bacteria on liver function decline induced by copper exposure. 3. The expression of IL-1 尾, IL-6 and IL-10 m RNA increased in low copper group, suggesting that low dose copper exposure could induce inflammatory injury in rat liver. 4. IL-1 尾, IL-6 and TNF- 伪 m RNA decreased significantly after the intervention of lactic acid bacteria, suggesting that the protective effect of lactic acid bacteria on liver injury induced by copper exposure may be related to the release of inflammatory cytokines.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R114
本文编号：2391661
[Abstract]:Objective: 1. To investigate the effects of low dose copper exposure on liver function and hepatic inflammatory factors in rats. 2. To investigate the effect of lactic acid bacteria (LAB) on liver function and inflammatory cytokine expression induced by copper exposure in rats. Methods: after 7 days of adaptive feeding, rats were randomly divided into 8 groups according to their body weight: high copper group (14mg/kg), medium copper group (3.5mg/kg group), low copper group (0.875mg/kg group) and high copper lactobacillus group (14mg/kg lactobacillus group). Medium copper lactobacillus group (3.5mg/kg group), low copper lactobacillus group (0.875mg/kg group lactic acid bacteria group), lactic acid bacteria group and blank control group, each group in accordance with the corresponding dose of continuous gastric administration of copper sulfate and lactic acid bacteria for 30 days. 1. At the end of the experiment, blood was taken from the abdominal aorta of the rats. After the rats were killed, the liver, spleen and thymus were taken and the indexes of liver, spleen and thymus were observed. 2. The liver tissue was embedded in paraffin, sectioned and stained with HE, and the pathological changes of liver were observed by microscope. 3. Serum proteins related to liver function: glutamate-pyruvic transaminase (ALT), aspartate aminotransferase (AST) and albumin (ALB), serum copper (Cu) in rats by 4.ICP-MS (inductively coupled plasma mass spectrometer). The contents of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in liver tissues were determined. Interleukin-1 尾 (IL-1 尾), interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12 (IL-12) in liver tissue were detected by 6.RT-PCR. The relative expression of superoxide dismutase and tumor necrosis factor 伪 TNF- 伪 m RNA. Results: 1. There was no significant difference in liver, spleen and thymus index among groups (P0.05). By observing the pathological sections of liver tissue, it was found that the liver tissues of high copper group, middle copper group and low copper group all had different degrees of damage under copper treatment, among which the high copper group was the most obvious. The liver injury in high copper group was improved to some extent after the intervention of lactobacillus. Compared with the control group, the AST,ALT of high copper group, medium copper group and low copper group was significantly higher (P0.05); ALB decreased significantly (P0.05); After the intervention of lactic acid bacteria, the ALB in copper exposure group increased significantly (P0.05), but only in medium copper group, AST and ALT were significantly improved in low copper group (P0.05). Compared with the control group, the levels of serum copper in different doses of copper exposure group were significantly higher (P0.05); after the intervention of lactic acid bacteria, the serum copper level of the three groups decreased, but there was no statistical significance (P0.05). Compared with the control group, the MDA content in the copper exposure group was significantly higher than that in the control group (P0.05), but the SOD activity and SOD m RNA decreased significantly in the high copper group and the middle copper group (P0.05). After the intervention of lactic acid bacteria, the levels of SOD and MDA in high copper group and middle copper group recovered significantly (P0.05). The levels of TNF- 伪, IL-1 尾, IL-6 and IL-10 m RNA were increased by different doses of copper exposure, and IL-12 m RNA was also increased in high copper group (P0.05). TNF- 伪, IL-1 尾 and IL-6 m RNA were significantly decreased after lactobacillus intervention (P0.05). Conclusion: 1. Different doses of copper could cause significant changes in AST,ALT and ALB, which indicated that low dose copper exposure could induce the decrease of liver function in rats. 2. After the intervention of lactic acid bacteria, both AST,ALT and ALB were improved to some extent, suggesting the protective effect of lactic acid bacteria on liver function decline induced by copper exposure. 3. The expression of IL-1 尾, IL-6 and IL-10 m RNA increased in low copper group, suggesting that low dose copper exposure could induce inflammatory injury in rat liver. 4. IL-1 尾, IL-6 and TNF- 伪 m RNA decreased significantly after the intervention of lactic acid bacteria, suggesting that the protective effect of lactic acid bacteria on liver injury induced by copper exposure may be related to the release of inflammatory cytokines.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R114
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