白藜芦醇对成年期追赶生长大鼠脂肪组织脂质生成和储存能力的影响及机制研究
发布时间：2019-07-03 19:22
【摘要】：第一部分成年期追赶生长大鼠模型的建立及一般生物学特征观察 目的：建立成年期追赶生长大鼠模型并观察其一般生物学特征。 方法：采用体重因素随机区组设计,按体重区组将6周龄雄性SD大鼠24只随机分为4组(共2个时间点),4周时间点2组：热卡限制4周组(R4),正常饮食4周组(NC4)；12周时间点2组：正常饮食追赶生长8周组(RN8)和持续正常饮食12周组(NC12)。通过先热卡限制后恢复饮食的方法建立成年期追赶生长大鼠模型。实验过程中每天记录实验大鼠进食量及体重变化,实验结束时观察实验大鼠热卡摄入量、体重和双能X线吸收测量法DEXA (Dual Energy X-ray Absorptiometry)检测不同部位的脂肪含量。高胰岛素-正糖钳夹实验评估追赶生长大鼠系统胰岛素敏感性及血糖、血清胰岛素水平检测。 结果： 1、实验动物实际能量摄入在限食期间约为NC组的60%,恢复饮食后基本达到NC组水平。 2、RN组在热卡限制期生长受抑,体重明显下降。恢复饮食后,体重增加迅速,但实验结束时仍未达NC组水平。 3、DEXA检测结果示限食时躯干组织脂肪含量及躯干脂肪占全身脂肪比例较正常对照组均下降,开放饮食后,RN8组全身、躯干脂肪含量及躯干脂肪比例均显著高于NC组。 4、高胰岛素-正葡萄糖钳夹实验显示RN组存在严重的系统胰岛素抵抗。R4组血糖和胰岛素水平较NC4组无明显变化,RN8组血糖水平较NC12组变化不明显,但胰岛素水平显著升高。 结论：通过热卡限制后恢复饮食方法建立的成年期追赶生长大鼠模型表现为热卡限制期体重生长抑制,恢复饮食后体重增长迅速和明显的体脂含量及内脏脂肪比例升高,且存在显著地系统胰岛素抵抗。与人群中发生的追赶生长时所观察到的变化类似,可作为一种较为理想的研究追赶生长的模型。 目的：探讨成年期追赶生长脂肪组织脂质生成与储存能力的改变和胰岛素抵抗之间的可能机制。 方法：采用体重因素随机区组设计,按体重区组将6周龄雄性SD大鼠随机分为8组其中4组(即追赶生长组群RN)为热卡限制组(R4)和追赶生长2、4、8周组(热卡限制后开放正常饮食RN2、RN4、RN8)；另外4组为与之年龄匹配的正常饮食对照组(即NC组群,分别为NC4、NC6、NC8、NC12)。检测附睾和皮下脂肪组织PPAR-γ、 FSP27的表达水平,检测两种脂肪组织在热卡限制期末及追赶生长8周结束时(R4、RN8)两个时间点的高胰岛素-正糖钳夹实验过程中2-脱氧-D葡萄糖摄取、血清甘油三酯、游离脂肪酸、脂联素及肿瘤坏死因子-α水平。 结果： 1、与NC4组相比,R4组附睾脂肪PPAR-γ的表达显著增加,而FSP27的表达明显下降；R4组皮下脂肪组织PPAR-γ的表达亦显著增加,但FSP27无明显变化。 2、与NC4组相比,R4组附睾脂肪和皮下脂肪组织2-脱氧-D葡萄糖摄取率轻度增加,但均未达到统计学意义。 3、R4组血清游离脂肪酸、脂联素水平较NC4组明显升高,甘油三酯、肿瘤坏死因子-α变化不显著。 4、附睾脂肪组织的PPAR-γ的表达水平在RN2、RN4、RN8组均明显增加,FSP27的mRNA表达水平在RN2组低于NC6组,在RN4组与NC8组无统计学差异,在RN8组明显高于NC12组。 5、皮下脂肪组织的PPAR-γ和FSP27表达水平在RN2、RN4、RN8组均无明显差异。 6、RN8组附睾脂肪组织2-脱氧-D葡萄糖摄取较NC12组显著增加,而皮下脂肪组织变化不明显。 7、RN8组血清游离脂肪酸、甘油三酯、肿瘤坏死因子-α水平均显著升高,而脂联素水平则明显降低。 结论：限食阶段内脏脂肪组织脂质生成和储存能力改变的不一致导致恢复饮食后脂质储存能力相对不足,加之生脂原料增加,使得过多的脂质不能有效的储存,脂质外溢,炎症因子释放增加,最终导致系统胰岛素抵抗的形成。皮下脂肪组织在该过程中变化不显著。因此,内脏脂肪的脂质生成和储存能力的失衡可能是成年期追赶生长胰岛素抵抗形成的重要原因。 目的：评估白藜芦醇在成年期追赶生长大鼠模型中对脂肪组织脂质生成和储存能力的影响。 方法：分组方法同第一部分,在4周和12周时间点各增加1组分别为R4+白藜芦醇、RN8+白藜芦醇(R4E、RN8E)组,即选取热卡限制结束(R4)及恢复饮食追赶生长结束(RN8)两个时间点作为观察白藜芦醇干预效果的研究。每天进行一定剂量白藜芦醇灌胃治疗,其余4组(NC4、R4、NC12和RN8)均进行相同剂量生理盐水灌胃治疗。记录白藜芦醇干预组的体重,DEXA检测干预组体脂含量,高胰岛素-正糖钳夹实验评估白藜芦醇对追赶生长大鼠(R4E和RN8E组)系统胰岛素敏感性的影响；检测4周组和8周组干预前后附睾和皮下脂肪组织SIRT-1的表达水平及活性；检测干预后附睾和皮下脂肪组织PPAR-γ和FSP27的表达水平、高胰岛素-正糖钳夹实验过程中2-脱氧-D葡萄糖摄取率、脂肪细胞的形态学改变；检测12周组附睾脂肪组织AKT及磷酸化AKT的表达；检测干预后血清甘油三酯、游离脂肪酸、脂联素及肿瘤坏死因子-α水平。 结果： 1、R4E和RN8E组全身组织脂肪含量、躯干组织脂肪含量、躯干脂肪占全身脂肪比例,均较干预前明显下降,达到甚至低于NC组。体重较干预前仅轻度下降,与RN组相比无统计学意义。 2、白藜芦醇对R4E组系统胰岛素敏感性、空腹血糖及空腹胰岛素水平无明显影响；RN8E组胰岛素敏感性和空腹胰岛素显著改善,基本恢复至正常水平,但对血糖水平影响不明显。 3、4周组中SIRT-1的表达和活性与NC4组相比,两种脂肪组织在R4组均明显升高；在12周组中与NC12组相比,两种脂肪组织在RN8组均明显降低；白藜芦醇干预后,两种组织在R4E、RN8E组SIRT-1的表达和活性超过或达到正常对照组。 4、经白藜芦醇干预后,附睾脂肪组织R4E、RN8E组FSP27的mRNA表达水平较干预前显著增加,超过NC组；PPAR-γ的表达明显下降,接近或低于NC组；2-脱氧-D葡萄糖摄取显著增加,高于NC组；RN8脂肪细胞组形状不规则,直径变大,胞膜不完整；RN8E组脂肪细胞形态学趋于正常。 5、与NC12组相比,RN8组大鼠附睾脂肪Ser473PKB磷酸化程度无显著变化,Thr308PKB磷酸化程度明显升高；白藜芦醇干预后,Ser473和Thr308PKB磷酸化程度均明显高于NC12及RN8组。 6、经白藜芦醇干预后,皮下脂肪组织R4E、RN8E组PPAR-γ的表达水平较干预前显著降低,接近或低于NC组；2-脱氧-D葡萄糖摄取显著增加,高于NC组；脂肪细胞在RN8、RN8E组形态学变化不明显。 7、R4E组和RN8E组血清游离脂肪酸、甘油三酯、肿瘤坏死因子-α均较干预前有所下降,在RN8E组下降趋势更为明显；脂联素水平则较干预前显著升高,恢复至NC组水平。 结论：白藜芦醇可抑制追赶生长过程中脂质的生成能力,提升脂质的储存能力,在一定程度上使二者改变接近平衡,并且保护脂肪细胞的完整性,减少脂质外溢及炎症因子释放,从而从本质上改善追赶生长所致的系统胰岛素抵抗。
[Abstract]:The establishment and general biological characteristics of the model of the first part after-one-year follow-up growth in rats Objective: To establish a rat model of catching and growing in adulthood and to observe its general biology. Methods: The 24-week-old male SD rats were randomly divided into 4 groups (2 time points) and 4 weeks at the time of 4 weeks:4 weeks (4 weeks),4 weeks (NC4) and 12 weeks in the body weight group. Point 2: normal diet followed up for 8 weeks (RN8) and continued normal diet for 12 weeks (NC 12) The method of recovering the diet after the restriction of the thermal card is established, and the growth of the one-year after-catch growth is established. Rat model was used to study the changes of feeding amount and body weight of the experimental rats per day during the course of the experiment. The experiment was conducted to observe the caloric intake, body weight and dual energy X-ray absorptiometry (DEXA (Dual Energy X-ray Absorbance) of the experimental rats. Determination of insulin sensitivity and blood glucose and serum insulin water in rats after catching and growing in rats with high insulin-positive sugar flat detection Results:1. The actual energy intake of the experimental animals was about 60% of the NC group during the food-limiting period. The level of the NC group is reached.2, the RN group is grown during the thermal card limit period. The body weight was significantly reduced. After the recovery of the diet, the weight gain was rapid, but the experiment was over The level of the NC group was still not reached.3. The results of DEXA test showed that the body fat content and the body fat in the body were lower than that in the normal control group. After the open diet, the body fat content and the trunk fat of the whole body, trunk and body of the RN8 group were decreased. The ratio of the two groups was significantly higher than that of the NC group.4. The experiment of high insulin-positive glucose clamp showed that the RN There was a serious systemic insulin resistance in the group. There was no significant change in the blood glucose and insulin levels in the group R4, and the level of blood glucose in the RN8 group was less than that of the N12 group. The results showed that the growth of body weight and body fat content and the proportion of visceral fat increased rapidly and the body fat content of the rats after the recovery of the diet was increased rapidly and the body fat content and the visceral fat ratio were increased after the diet was restored by the hot-card restriction. high, and there is a significant system insulin resistance, similar to that observed in the follow-up growth that occur in the population, An ideal model for the study of the catch-up and growth of adipose tissue was studied. Objective: To study the formation and storage of lipid in the growth adipose tissue during the period of adult-year follow-up. Methods:6-week-old male SD rats were randomly divided into 8 groups by weight-factor random-zone design,4 groups (i.e., catch-up group RN) were heat-card-limiting group (R4) and after-catch growth of 2,4, and 8-week group (thermal card limitation). The normal diet (RN2, RN4, RN8) and the normal diet control group (i.e., NC group, sub-group) matched with the age of the normal diet were group 4. The expression levels of PPAR-1 and FSP27 in the epididymis and the subcutaneous fat tissue were detected, and the high insulin-positive ratio of the two adipose tissues at two time points at the end of the caloric restriction period and at the end of the after-catch growth of 8 weeks (R4, RN8) was detected. 2-deoxy-D glucose uptake, serum triglyceride, and swim in the course of the sugar-jaw experiment off fat Results:1. The expression of PPAR-1 in the epididymis of R4 group increased significantly compared with that of the NC4 group, while the expression of FSP27 decreased significantly. The expression of R-1 was also significantly increased, but there was no significant change in FSP27. In comparison with the NC4 group, the epididymal fat and the subcutaneous fat tissue of the group R4 were 2- The results showed that the levels of free fatty acid and adiponectin in group 3 and R4 were higher than those in group 3 and 4. The expression of PPAR-1 in the adipose tissue of the epididymis increased significantly in the RN2, RN4 and RN8 groups, and the expression level of FSP27 was lower than that of the NC6 group in the RN2 group. There was no statistical difference between the N4 group and the NC8 group, and the PPAR of the subcutaneous fat tissue was significantly higher in the RN8 group than in the NC12 group. The expression levels of-1 and FSP27 were not significantly different in RN2, RN4 and RN8 groups. -Deoxy-D glucose uptake increased significantly in the NC12 group, while the changes in the subcutaneous fat tissue were not significant. Conclusion: The changes of lipid production and storage capacity of visceral fat tissue in the limited-feeding stage are not consistent with the change of lipid production and storage capacity of the visceral fat tissue in the limiting stage, and the lipid storage capacity after the recovery of the diet is relatively low. In addition, the raw material of the raw fat is increased, so that the excessive fat It is not effective in storage, lipid spillovers, and inflammation. The increase of the sub-release results in the formation of the system's insulin resistance. The subcutaneous fat tissue changes in this process. Therefore, the visceral organs Imbalance of the lipid generation and storage capacity of fat may be an important cause of the formation of the growth of insulin resistance in adulthood. Objective: To evaluate the effect of the white-calcitol on the lipid formation and storage capacity of adipose tissue in the growth rat model after the adulthood. Methods: The grouping method is the same as the first part, and the effect of the method is 4 weeks and 4 weeks. At the 12-week time point, group 1 was the group R4 + white, and the group of R4E and RN8E respectively, that is, the heat-card-limiting junction was selected. A study of the effect of the two time points of the beam (R4) and the recovery diet after the end of the growth (RN8) was used as a study to observe the effect of the intervention of the aloe. The rats were treated with the same dose of saline in the other four groups (NC4, R4, NC12, and RN8). To evaluate the effect of the white-calcitol on the insulin sensitivity of the rats (R4E and RN8E) in the follow-up growth of rats (R4E and RN8E). The expression level and activity of SIRT-1 in the epididymis and the subcutaneous fat tissue before and after the intervention of the 4-week and 8-week groups were examined. The expression level of R-1 and FSP27, the rate of 2-deoxy-D glucose uptake and the morphological changes of fat cells during the experiment of high insulin-positive sugar clamp, and the detection of AKT and P in the epididymis adipose tissue in the 12-week group. Acidification A Expression of KT, serum triglyceride, free fatty acid, adiponectin and tumor necrosis factor-1 level after intervention. Results:1, R4E and RN8E were all body tissues. Fat content, body fat content, body fat proportion of body fat There was no statistical significance in the insulin sensitivity, fasting blood glucose and fasting insulin levels in the R4E group. The expression and activity of SIRT-1 in group 3 and 4 were significantly higher in group 4 than in the NC4 group. In the 12-week group, the two adipose tissues were in the RN8 group compared with the N12 group. The expression and activity of the two tissues in the R4E and RN8E group SIRT-1 exceeded or reached the normal control group. And the expression of PPAR-1 is significantly lower, close to or lower than that of NC group, and 2-deoxy-D glucose uptake is obvious. The increase was higher than that of the NC group, the shape of the group of RN8 adipocytes was irregular, the diameter of the cell was large, the cell membrane was not complete, the morphology of the fat cells in the RN8E group tended to be normal.5. Compared with the NC12 group, there was no significant change in the phosphorylation of the fat Ser473PKB in the epididymis of the RN8 group, Thr308PKB The level of phosphorylation of Ser473 and Thr308PKB was significantly higher than that of NC12 and RN8 groups. Low, close to or lower than NC group;2-deoxy-D glucose uptake increased significantly, which was higher than that of NC group; the morphological changes of fat cells in RN8 and RN8E group were not obvious. And the level of adiponectin decreased significantly before intervention and returned to the level of NC group. The ability to produce, to improve the storage capacity of the lipid, to a certain extent
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R285.5
本文编号：2509608
[Abstract]:The establishment and general biological characteristics of the model of the first part after-one-year follow-up growth in rats Objective: To establish a rat model of catching and growing in adulthood and to observe its general biology. Methods: The 24-week-old male SD rats were randomly divided into 4 groups (2 time points) and 4 weeks at the time of 4 weeks:4 weeks (4 weeks),4 weeks (NC4) and 12 weeks in the body weight group. Point 2: normal diet followed up for 8 weeks (RN8) and continued normal diet for 12 weeks (NC 12) The method of recovering the diet after the restriction of the thermal card is established, and the growth of the one-year after-catch growth is established. Rat model was used to study the changes of feeding amount and body weight of the experimental rats per day during the course of the experiment. The experiment was conducted to observe the caloric intake, body weight and dual energy X-ray absorptiometry (DEXA (Dual Energy X-ray Absorbance) of the experimental rats. Determination of insulin sensitivity and blood glucose and serum insulin water in rats after catching and growing in rats with high insulin-positive sugar flat detection Results:1. The actual energy intake of the experimental animals was about 60% of the NC group during the food-limiting period. The level of the NC group is reached.2, the RN group is grown during the thermal card limit period. The body weight was significantly reduced. After the recovery of the diet, the weight gain was rapid, but the experiment was over The level of the NC group was still not reached.3. The results of DEXA test showed that the body fat content and the body fat in the body were lower than that in the normal control group. After the open diet, the body fat content and the trunk fat of the whole body, trunk and body of the RN8 group were decreased. The ratio of the two groups was significantly higher than that of the NC group.4. The experiment of high insulin-positive glucose clamp showed that the RN There was a serious systemic insulin resistance in the group. There was no significant change in the blood glucose and insulin levels in the group R4, and the level of blood glucose in the RN8 group was less than that of the N12 group. The results showed that the growth of body weight and body fat content and the proportion of visceral fat increased rapidly and the body fat content of the rats after the recovery of the diet was increased rapidly and the body fat content and the visceral fat ratio were increased after the diet was restored by the hot-card restriction. high, and there is a significant system insulin resistance, similar to that observed in the follow-up growth that occur in the population, An ideal model for the study of the catch-up and growth of adipose tissue was studied. Objective: To study the formation and storage of lipid in the growth adipose tissue during the period of adult-year follow-up. Methods:6-week-old male SD rats were randomly divided into 8 groups by weight-factor random-zone design,4 groups (i.e., catch-up group RN) were heat-card-limiting group (R4) and after-catch growth of 2,4, and 8-week group (thermal card limitation). The normal diet (RN2, RN4, RN8) and the normal diet control group (i.e., NC group, sub-group) matched with the age of the normal diet were group 4. The expression levels of PPAR-1 and FSP27 in the epididymis and the subcutaneous fat tissue were detected, and the high insulin-positive ratio of the two adipose tissues at two time points at the end of the caloric restriction period and at the end of the after-catch growth of 8 weeks (R4, RN8) was detected. 2-deoxy-D glucose uptake, serum triglyceride, and swim in the course of the sugar-jaw experiment off fat Results:1. The expression of PPAR-1 in the epididymis of R4 group increased significantly compared with that of the NC4 group, while the expression of FSP27 decreased significantly. The expression of R-1 was also significantly increased, but there was no significant change in FSP27. In comparison with the NC4 group, the epididymal fat and the subcutaneous fat tissue of the group R4 were 2- The results showed that the levels of free fatty acid and adiponectin in group 3 and R4 were higher than those in group 3 and 4. The expression of PPAR-1 in the adipose tissue of the epididymis increased significantly in the RN2, RN4 and RN8 groups, and the expression level of FSP27 was lower than that of the NC6 group in the RN2 group. There was no statistical difference between the N4 group and the NC8 group, and the PPAR of the subcutaneous fat tissue was significantly higher in the RN8 group than in the NC12 group. The expression levels of-1 and FSP27 were not significantly different in RN2, RN4 and RN8 groups. -Deoxy-D glucose uptake increased significantly in the NC12 group, while the changes in the subcutaneous fat tissue were not significant. Conclusion: The changes of lipid production and storage capacity of visceral fat tissue in the limited-feeding stage are not consistent with the change of lipid production and storage capacity of the visceral fat tissue in the limiting stage, and the lipid storage capacity after the recovery of the diet is relatively low. In addition, the raw material of the raw fat is increased, so that the excessive fat It is not effective in storage, lipid spillovers, and inflammation. The increase of the sub-release results in the formation of the system's insulin resistance. The subcutaneous fat tissue changes in this process. Therefore, the visceral organs Imbalance of the lipid generation and storage capacity of fat may be an important cause of the formation of the growth of insulin resistance in adulthood. Objective: To evaluate the effect of the white-calcitol on the lipid formation and storage capacity of adipose tissue in the growth rat model after the adulthood. Methods: The grouping method is the same as the first part, and the effect of the method is 4 weeks and 4 weeks. At the 12-week time point, group 1 was the group R4 + white, and the group of R4E and RN8E respectively, that is, the heat-card-limiting junction was selected. A study of the effect of the two time points of the beam (R4) and the recovery diet after the end of the growth (RN8) was used as a study to observe the effect of the intervention of the aloe. The rats were treated with the same dose of saline in the other four groups (NC4, R4, NC12, and RN8). To evaluate the effect of the white-calcitol on the insulin sensitivity of the rats (R4E and RN8E) in the follow-up growth of rats (R4E and RN8E). The expression level and activity of SIRT-1 in the epididymis and the subcutaneous fat tissue before and after the intervention of the 4-week and 8-week groups were examined. The expression level of R-1 and FSP27, the rate of 2-deoxy-D glucose uptake and the morphological changes of fat cells during the experiment of high insulin-positive sugar clamp, and the detection of AKT and P in the epididymis adipose tissue in the 12-week group. Acidification A Expression of KT, serum triglyceride, free fatty acid, adiponectin and tumor necrosis factor-1 level after intervention. Results:1, R4E and RN8E were all body tissues. Fat content, body fat content, body fat proportion of body fat There was no statistical significance in the insulin sensitivity, fasting blood glucose and fasting insulin levels in the R4E group. The expression and activity of SIRT-1 in group 3 and 4 were significantly higher in group 4 than in the NC4 group. In the 12-week group, the two adipose tissues were in the RN8 group compared with the N12 group. The expression and activity of the two tissues in the R4E and RN8E group SIRT-1 exceeded or reached the normal control group. And the expression of PPAR-1 is significantly lower, close to or lower than that of NC group, and 2-deoxy-D glucose uptake is obvious. The increase was higher than that of the NC group, the shape of the group of RN8 adipocytes was irregular, the diameter of the cell was large, the cell membrane was not complete, the morphology of the fat cells in the RN8E group tended to be normal.5. Compared with the NC12 group, there was no significant change in the phosphorylation of the fat Ser473PKB in the epididymis of the RN8 group, Thr308PKB The level of phosphorylation of Ser473 and Thr308PKB was significantly higher than that of NC12 and RN8 groups. Low, close to or lower than NC group;2-deoxy-D glucose uptake increased significantly, which was higher than that of NC group; the morphological changes of fat cells in RN8 and RN8E group were not obvious. And the level of adiponectin decreased significantly before intervention and returned to the level of NC group. The ability to produce, to improve the storage capacity of the lipid, to a certain extent
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R285.5
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相关期刊论文 前2条
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