基于P38MAPK通路探讨祛痰活血颗粒调节NAFLD模型大鼠脂肪组织AQP7表达的机制

发布时间：2018-06-20 10:09

本文选题：NAFLD + AQP7　；参考：《西南医科大学》2017年硕士论文

【摘要】：目的:观察祛痰活血颗粒治疗NAFLD模型大鼠后脂肪组织AQP7、P38MAPK的表达变化,探讨其治疗NAFLD的作用机制。方法:选择40只体重在180~220g清洁级雄性SD大鼠,普通饲料适应性饲养1周后,按随机数字表法分成正常组(6只)、模型组(34只);正常组给予普通饲料饲喂,模型组采用高脂饮食诱导的方法复制NAFLD大鼠模型,于第10周开始,每隔2周随机抽取1~2只大鼠处死,取肝脏做病理染色,观察其病理变化,确认造模成功后,将剩余模型组再随机分为模型对照组6只,中药高剂量组(6只)、中剂量组(6只)、低剂量组(6只),抑制剂组(6只),各中药组灌喂祛痰活血颗粒,抑制剂组腹腔注射P38MAPK抑制剂(SB203580)干预,连续干预4周后处死大鼠,光镜下观察肝脏HE染色及油红O染色变化,测定各组大鼠血清TG、TC、AST、ALT,肝脏TG、FFA,肝脏指数和脂肪组织AQP7及P38MAPK基因及蛋白表达。结果:1、病理组织HE及油红O染色示:与正常组比较,模型组大鼠肝细胞内脂滴数量多,肝脏脂肪变性明显,差异有统计学意义(P0.05);与模型组比较,中药组与抑制剂组大鼠肝细胞内脂滴数量减少,肝脏脂肪变性程度明显减轻(P0.05);2、血清生化指标示:与正常组比较,模型组大鼠的血清TG、TC、AST、ALT明显升高;与模型组比较,各中药组及抑制剂组大鼠的血清TG、TC、AST、ALT明显降低(P0.05);3、肝脏tg、ffa及肝脏指数示:与正常组比较,模型组大鼠的肝脏tg、ffa和肝脏指数明显升高;与模型组比较,中药组及抑制剂组大鼠的肝脏tg、ffa和肝脏指数明显降低(p0.05);4、realtime-pcr检测结果示:与正常组比较,模型组大鼠脂肪组织中aqp7mrna表达降低,p38mapkmrna表达升高(p0.05);与模型组比较,各中药组和抑制剂组大鼠脂肪组织中aqp7mrna表达明显升高,以中药高剂量组效果最好;p38mapkmrna表达明显降低,以抑制剂组效果最明显,中药高剂量组效果次之(p0.05);中药低、中剂量组大鼠间的aqp7和p38mapkmrna表达差异无统计学意义(p0.05);5、蛋白检测结果示:与正常组比较,模型组大鼠脂肪组织中aqp7蛋白表达明显降低,p-p38mapk蛋白表达明显升高(p0.05);与模型组比较,各中药组和抑制剂组大鼠脂肪组织中aqp7蛋白表达明显升高,以中药高剂量组效果最好;p-p38mapk蛋白表达明显降低,以抑制剂组效果最明显,中药高剂量组效果次之(p0.05);中药低、中剂量组大鼠间aqp7和p-p38mapk蛋白表达差异无统计学意义(p0.05);各组大鼠脂肪组织总p38mapk蛋白无明显差异(p0.05)。结论:1、祛痰活血颗粒可改善nafld的病理结构,减轻nafld大鼠肝脏脂肪变性程度及肝脏炎症;2、祛痰活血颗粒可降低nafld大鼠肝脏指数、血清tg、tc、ast、alt及肝脏tg、ffa;3、祛痰活血颗粒可能通过抑制脂肪组织中p38mapk活化、上调aqp7表达,增加甘油入血代谢,减少进入肝脏的甘油及ffa,从而减少肝脏脂肪蓄积,减轻肝脂脂肪变性程度;4、脂肪组织中p38mapk通路可能参与抑制AQP7的表达,起到调控甘油及脂肪代谢,从而间接改善NAFLD。
[Abstract]:Objective: to observe the changes of AQP7 and P38 MAPK expression in adipose tissue of NAFLD rats treated with Quphan Huoxue granule and to explore the mechanism of its therapeutic effect on NAFLD. Methods: forty male SD rats weighing 180 ~ 220g were selected and fed with normal diet for one week, then were randomly divided into normal group (n = 6), model group (n = 34) and normal group (n = 34). In the model group, NAFLD rat model was induced by high-fat diet. From the 10th week on, 2 rats were randomly selected every 2 weeks to be killed. The liver was taken for pathological staining, the pathological changes were observed, and the model was established successfully. The remaining model group was randomly divided into two groups: control group (n = 6), high dose group (n = 6), middle dose group (n = 6), low dose group (n = 6) and inhibitor group (n = 6). The rats in the inhibitor group were treated with P38 MAPK inhibitor SB203580 intraperitoneally. The rats were killed after 4 weeks of continuous intervention. The liver HE staining and oil red O staining were observed under light microscope. The levels of serum TGG, liver TGfFA, AQP7 and P38 MAPK gene and protein expression in adipose tissue were measured. Results: the pathological tissues were stained with HE and oil red O: compared with the normal group, the rats in the model group had more lipid droplets and steatosis in the liver, the difference was statistically significant (P 0.05), and compared with the model group, there was no significant difference between the model group and the model group. The number of lipid droplets in liver cells in the Chinese medicine group and the inhibitor group were decreased, and the degree of steatosis in the liver was significantly reduced (P 0.05). The serum biochemical indexes showed that compared with the normal group, the serum TGN TCASTN alt of the model group was significantly higher than that of the model group, and that of the model group was significantly higher than that of the model group. The serum levels of TGG, TCC, AST and liver index of rats in the traditional Chinese medicine group and the inhibitor group were significantly lower than those in the control group, and the liver tgffa and liver index in the model group were significantly higher than those in the model group, and in comparison with the model group, the liver index of the model group was significantly higher than that of the control group, and that of the model group was significantly higher than that of the control group. Compared with the normal group, the expression of aqp7mrna in the adipose tissue of the model group decreased significantly, and the expression of p38 mapkmrna increased in the model group, and compared with the model group, the liver index of the rats in the traditional Chinese medicine group and the inhibitor group was significantly decreased, and that in the model group was significantly higher than that in the model group, the results showed that: compared with the model group, the expression of aqp7mrna in the adipose tissue of the model group was significantly lower than that in the control group. The expression of aqp7mrna in adipose tissue of rats in each Chinese medicine group and inhibitor group was significantly increased, and the expression of p38 mapkrna was the best in the high dose group, especially in the inhibitor group, followed by p0.05 in the high dose group, and low in the Chinese medicine group, and the expression of p38mapkmrna in the high dose group was lower than that in the control group. There was no significant difference in the expression of aqp7 and p38mapkmrna between the middle dose group and the middle dose group. The results of protein detection showed that the expression of aqp7 protein in adipose tissue of the model group was significantly lower than that in the normal group, and the expression of p-p38 mapk protein was significantly increased in the model group, while that in the model group was significantly higher than that in the model group. The expression of aqp7 protein in adipose tissue of rats in each Chinese medicine group and inhibitor group was significantly increased, and the expression of p-p38 mapk protein was the best in high dose group, especially in inhibitor group, followed by p0.05 in high dose group and low in Chinese medicine group. There was no significant difference in the expression of aqp7 and p-p38mapk protein between the middle dose group and the adipose tissue of the rats in each group (p 0.05), but there was no significant difference in the total p38mapk protein in the adipose tissue of the rats in each group (P 0.05). Conclusion: W1, Quphan Huoxue granule can improve the pathological structure of nafld, reduce the degree of hepatic steatosis and liver inflammation in nafld rats. Quphlegm Huoxue granule can reduce the liver index of nafld rats. Serum TGP tcastan and liver tgctctcastastan and liver TGP 3. Quphlegm Huoxue granule may reduce hepatic fat accumulation by inhibiting p38mapk activation in adipose tissue, upregulating aqp7 expression, increasing glycerol metabolism, reducing glycerol and FFA entering the liver, and reducing liver fat accumulation. In adipose tissue, p38mapk pathway may be involved in inhibiting AQP7 expression, regulating glycerol and fat metabolism, and thus indirectly improving NAFLD.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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