间歇性低氧对大鼠肝脏的损伤和脂肪因子chemerin表达的影响及复氧的干预作用

发布时间：2018-04-27 12:58

本文选题：间歇性低氧 + 阻塞性睡眠呼吸暂停低通气综合征　；参考：《兰州大学》2017年硕士论文

【摘要】：目的通过研究间歇性低氧(IH)对大鼠肝脏的损伤及脂肪因子chemerin表达的影响,探讨非酒精性脂肪性肝病(NAFLD)与脂肪因子chemerin表达的内在关系,并研究复氧的干预作用,为临床上OSAHS及其相关代谢性并发症的进一步研究及防治提供新的思路。方法选取48只健康的雄性Wistar大鼠,根据随机数字表法随机分为4组(n=12):常氧+普通饮食组(NC+ND组)、常氧+高脂饮食组(NC+HFD组)、间歇低氧+普通饮食组(IH+ND组)、间歇低氧+高脂饮食组(IH+HFD组)。普通饮食组给予基础饲料喂养,高脂饮食组予以高脂饲料喂养。IH组置于8h/d的间歇性低氧环境中,同时NC组给予间歇压缩空气。实验6周末各组处死一半大鼠,称量体重、肝湿重,计算肝指数,采集动脉血及肝组织标本,使用全自动生化分析仪检测大鼠肝酶、血脂水平,空腹血糖(FPG)通过葡萄糖氧化酶法检测,空腹胰岛素(FINS)使用放射免疫法测定,用胰岛素敏感指数(ISI)及胰岛素抵抗指数(IRI)系统评价胰岛素抵抗,光镜、透射电镜观察肝脏组织形态学及超微结构改变,ELISA(双抗体夹心法)检测大鼠血清及肝组织chemerin的表达。余IH组大鼠予以复氧干预2周,普通饮食组继续给予基础饲料喂养,高脂饮食组予以高脂饲料喂养,实验8周末处死剩余大鼠,各检测指标及检测方法同上所述。结果1.与NC+ND组相比,IH+ND组大鼠血清ALT、AST、FPG、FINS、IRI、TC、TG、LDL-C均显著升高;与NC+HFD组相比,IH+HFD组大鼠血清AST、FINS、IRI水平升高;与IH+ND组相比,IH+HFD组大鼠血清AST、FPG、TC均升高,LDL-C水平显著升高。余各项指标间未见明显差异。2.与NC+ND组相比,在光学显微镜和透射电子显微镜下,NC+HFD组、IH+ND组和IH+HFD组大鼠肝组织学特点均表现为非酒精性脂肪性肝病:NC+HFD组表现为单纯性脂肪肝;IH+ND组表现为脂肪性肝炎,IH+HFD组损伤表现均明显重于IH+ND组和NC+HFD组,表现为脂肪性肝炎甚至轻度肝脏纤维化。3.与NC+ND组相比,IH+ND组大鼠血清chemerin表达水平显著升高;与NC+HFD组和IH+ND组相比,IH+HFD组大鼠血清chemerin水平亦有显著表达。其余各项指标间未见明显统计学意义。4.大鼠血清chemerin表达水平与ALT(r=0.526)呈正相关,与AST(r=0.742)、FPG(r=0.751)、FINS(r=0.764)、IRI(r=0.765)、TC(r=0.791)、LDL-C(r=0.818)、体重(r=0.685)呈明显正相关,与ISI(r=-0.692)呈明显负相关。5.以是否发生OSAHS相关NAFLD为因变量,以肝重、体重、LBR、AST、ALT、FPG、FINS、IRI、ISI、TC、TG、LDL-C、血清chemerin及肝脏chemerin为自变量,行Logistic逐步回归分析后得出结果,血清chemerin的表达是发生OSAHS相关NAFLD的危险因素(OR=1.197,P0.01)。6.与复氧前相比,复氧后IH+ND组大鼠体重升高,IRI水平降低,血清FPG、血清chemerin表达水平显著降低,在光学显微镜和透射电子显微镜下,肝组织学特点仍表现为脂肪性肝炎;与复氧前相比,复氧后IH+HFD组大鼠血清FPG水平降低,血清chemerin表达水平显著降低,肝组织学特点亦仍表现为脂肪性肝炎甚至轻度肝脏纤维化。结论1.间歇性低氧可导致大鼠发生糖脂代谢紊乱及肝损伤,主要表现为NAFLD,在间歇性低氧合并高脂饮食的情况下,这种代谢紊乱与肝损伤更为严重。提示间歇性低氧可能是NAFLD发生与进展的独立危险因素。2.暴露于间歇性低氧条件下的大鼠发生OSAHS相关NAFLD时,血清chemerin水平呈高表达状态,肝脏chemerin水平无显著变化,行Logistic逐步回归分析结果显示,血清chemerin是发生NAFLD的危险因素,提示血清chemerin可作为早期预测OSAHS相关NAFLD的脂肪因子。3.两周的复氧干预治疗并不能引起大鼠肝脏病理表现和血清转氨酶水平的明显改观,但可以降低FPG水平,甚至减轻胰岛素抵抗,并使得血清chemerin表达水平显著下降,提示短期CPAP治疗无法有效改善OSAHS相关NAFLD,但在一定程度上可能会稳定和延缓其进展,血清chemerin可能作为未来治疗OSAHS相关NAFLD新的非侵入性标记物。
[Abstract]:Objective to study the effect of intermittent hypoxia (IH) on the liver injury and the expression of fat factor Chemerin in rats, to explore the intrinsic relationship between the expression of nonalcoholic fatty liver disease (NAFLD) and fat factor Chemerin, and to study the intervention of reoxygenation, and provide a new method for the further study and prevention of the clinical OSAHS and its related metabolic complications. Methods 48 healthy male Wistar rats were randomly divided into 4 groups (n=12): normal oxygen + ordinary diet group (group NC+ND), normal oxygen + high fat diet group (group NC+HFD), intermittent hypoxia + ordinary diet group (IH+ND group), intermittent hypoxia + high fat diet group (group IH+HFD). The ordinary diet group was given basic feed, high fat drink. The diet group was given high fat diet for.IH group to be placed in the intermittent hypoxia environment of 8h/d, while group NC was given intermittent compressed air. At the end of the 6 week, half of the rats were killed, weighing weight, liver wet weight, calculating liver index, collecting arterial blood and liver tissue specimens, using an automatic biochemical analyzer to detect rat liver enzyme, blood lipid level, fasting blood glucose (FPG Through the glucose oxidase assay, the fasting insulin (FINS) was measured by radioimmunoassay, insulin resistance index (ISI) and insulin resistance index (IRI) system were used to evaluate insulin resistance, light microscopy and transmission electron microscopy were used to observe the changes of the liver histomorphology and ultrastructure, and the ELISA (double antibody sandwich method) was used to detect the chemer in the serum and liver tissue of rats. The expression of in. The rats in group IH were treated with reoxygenation for 2 weeks. The normal diet group continued to feed the basal diet, the high fat diet group was fed with high fat diet, and the remaining rats were killed at the end of the 8 week. The results 1. were compared with the NC+ND group. The serum ALT, AST, FPG, FINS, IRI, TC, TG, LDL-C were all significant compared with the group IH+ND. The level of serum AST, FINS and IRI in group IH+HFD was higher than that in group NC+HFD. Compared with group IH+ND, the serum AST, FPG, TC of rats in group IH+HFD increased and LDL-C level increased significantly. The histological features were all nonalcoholic fatty liver disease: NC+HFD group showed simple fatty liver, IH+ND group showed fatty hepatitis, and group IH+HFD was significantly more serious than group IH+ND and NC+HFD group. The expression of.3. in fatty hepatitis and even mild liver fibrosis was compared with that of group NC+ND, and the serum Chemerin expression level of IH+ND group was significantly higher than that of group NC+ND. As compared with group NC+HFD and group IH+ND, the level of serum Chemerin in group IH+HFD was also significantly expressed. There was no significant statistical significance between the other indexes. The level of Chemerin expression in the serum of.4. rats was positively correlated with the ALT (r=0.526) and AST (r=0.742), FPG (r=0.751). 685) there was a significant positive correlation and a significant negative correlation with ISI (r=-0.692), which was negatively correlated with.5., with the occurrence of OSAHS related NAFLD as the dependent variable, with liver weight, weight, LBR, AST, ALT, FPG, FINS, IRI, ISI, serum and liver as independent variables. The risk factors (OR=1.197, P0.01).6. were compared with that before reoxygenation. After reoxygenation, the body weight of the rats in the IH+ND group, the IRI level, the serum FPG, the serum Chemerin expression level decreased significantly. The histological characteristics of the liver were still fatty hepatitis in the optical microscope and transmission electron microscope, and the serum FPG in IH+HFD group after reoxygenation was compared with the reoxygenation. The level of serum Chemerin expression decreased significantly and the liver histology was still characterized by fatty hepatitis and even mild liver fibrosis. Conclusion 1. intermittent hypoxia can lead to glucose and lipid metabolism disorder and liver injury in rats. The main manifestation is NAFLD. In the case of intermittent hypoxic combined with high fat diet, this metabolic disorder and liver disease It is suggested that intermittent hypoxia may be an independent risk factor for the occurrence and progression of NAFLD. When.2. exposure to intermittent hypoxia, the level of serum Chemerin is highly expressed, and the level of Chemerin in the liver is not significantly changed. The result of Logistic stepwise regression analysis shows that serum Chemerin is the hair of hair. The risk factors of birth NAFLD suggest that serum Chemerin can be used as an early prediction of OSAHS related NAFLD for.3. two weeks of reoxygenation, which can not cause a significant change in liver pathological manifestation and serum transaminase level, but can reduce the level of FPG, even reduce islet resistance, and make the level of serum Chemerin significant. Decline, suggesting that short term CPAP therapy does not effectively improve OSAHS related NAFLD, but may stabilize and delay its progress to a certain extent. Serum Chemerin may be a new noninvasive marker for the future treatment of OSAHS related NAFLD.

【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575;R766

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