电针对高脂喂养大鼠主动脉内皮细胞JNK蛋白的影响

发布时间：2018-10-24 20:26

【摘要】：目的:本次实验在前期实验研究的基础上,将高脂饮食诱导的胰岛素抵抗模型大鼠进行电针干预治疗,观察主动脉内皮细胞JNK蛋白的变化,探究并完善电针改善胰岛素抵抗状态的内在机制,同时也为临床治疗胰岛素抵抗相关疾病提供参考。方法:在SPF级实验室标准环境下,本次实验研究设立3个组,分别为空白组、模型组和电针组,实验动物选择雄性SD大鼠24只,随机分到3组,每组8只动物。空白组的实验大鼠选用普通饲料进行喂养,而模型组与电针组的实验大鼠则采用高脂饲料进行喂养。喂养12周后,检测三组实验模型大鼠的空腹胰岛素(FINS)和空腹血糖(FPG)数据并计算稳态模型评价的胰岛素抵抗指数(HOMA-IR),从而评价模型的建立是否成功。造模成功后,电针组的实验大鼠选择双侧内关穴、三阴交穴、足三里穴以及肾俞穴进行针刺干预治疗,其中双侧足三里穴和三阴交穴采用电针疗法进行干预,并继续以高脂饲料喂养。电针治疗选用疏密波(密波50 Hz,疏波2 Hz,频率转换10次/min),其强度以实验大鼠后肢轻微抖动为度,20 min/次,1次/d,持续4w。而空白组与模型组的实验大鼠则继续分别予普通饲料和高脂饲料进行喂养,仅采用与电针组相同的按压固定时间,其余不做干预。4周后,检测三组实验大鼠的空腹胰岛素(FINS)、空腹血糖(FPG)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)以及胰岛素抵抗指数(H0MA-IR)的数据并计算稳态模型评价的胰岛素抵抗指数(H0MA-IR)。采用Western-blotting方法检测主动脉内皮细胞JNK蛋白的表达及其磷酸化水平。结果:喂养12周后,电针组与模型组实验大鼠FPG、FINS以及H0MA-IR相比于空白组有显著升高(P＜0.0);电针治疗4周后,比较空白组与模型组结果:模型组大鼠FPG、FINS、TG、TC、以及LDL-C均有显著升高(P0.01),而HDL-C则明显降低(P0.01);模型组大鼠H0MA-IR也有显著升高(P0.01);此外,模型组大鼠的JNK蛋白的表达及其磷酸化水平比空白组大鼠也有显著增加(P＜0.01)。比较电针组和模型组的结果:电针组大鼠FPG、FINS、TG、TC、以及LDL-C均有显著降低(P＜0.01),而HDL-C则明显升高(P＜0.01);电针组大鼠H0MA-IR也有显著减低(P＜0.01)。此外,电针组大鼠的JNK蛋白的表达及其磷酸化水平比空白组大鼠也有显著降低(P＜0.01)。结论:高脂饮食喂养大鼠能够成功诱导实验大鼠胰岛素抵抗的状态,实现胰岛素抵抗模型的建立。电针疗法通过针刺三阴交、足三里、肾俞穴、内关,健脾补肾、祛瘀化痰通络,降低大鼠体内FPG、FINS、TG、TC以及LDL-C的含量,并提高HDL-C的含量,从而改善机体内糖代谢和脂代谢的紊乱状态;与此同时,下调主动脉内皮细胞JNK蛋白的表达及其磷酸化水平,恢复胰岛素信号传导通路,从而改善模型大鼠胰岛素抵抗状态,且模型大鼠对胰岛素的敏感性也有增强。
[Abstract]:Aim: to observe the changes of JNK protein in aortic endothelial cells by electroacupuncture intervention on insulin resistance model rats induced by high fat diet on the basis of previous experimental study. To explore and improve the internal mechanism of electroacupuncture to improve insulin resistance, and to provide reference for clinical treatment of insulin resistance related diseases. Methods: under the standard environment of SPF laboratory, three groups were established in this experiment, namely blank group, model group and electroacupuncture group. 24 male SD rats were randomly divided into 3 groups with 8 animals in each group. The rats in the blank group were fed with common diet, while the rats in the model group and electroacupuncture group were fed with high fat diet. After 12 weeks of feeding, fasting insulin (FINS) and fasting blood glucose (FPG) were measured in three groups of experimental rats. The insulin resistance index (HOMA-IR) of steady-state model was calculated to evaluate the success of the model. After the successful establishment of the model, the experimental rats in the electroacupuncture group were treated with acupuncture intervention at the points of bilateral Neiguan, Sanyinjiao, Zusanli and Shenshu, among which the bilateral Zusanli and Sanyinjiao were treated by electroacupuncture. And continue to feed on high fat feed. In electroacupuncture treatment, the density wave (density wave 50 Hz, sparse wave 2 Hz, frequency conversion 10 times / min),) was used as the intensity of the experimental rat hind limb slight shaking, 20 min/ times, once a day for 4 wks. However, the rats in the blank group and the model group continued to be fed with normal diet and high fat diet respectively, only the same pressing time as the electroacupuncture group, and the rest did not intervene after 4 weeks. Fasting insulin, (FINS), fasting blood glucose, (FPG), total cholesterol, (TC), triglyceride, (TG), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C) and insulin resistance index (H0MA-IR) were measured and the insulin resistance index (H0MA-IR) evaluated by steady-state model was calculated. The expression and phosphorylation of JNK protein in aortic endothelial cells were detected by Western-blotting. Results: after 12 weeks of feeding, the FPG,FINS and H0MA-IR of the electroacupuncture group and the model group were significantly higher than those of the blank group (P < 0. 0. 0). The results of comparison between the blank group and the model group showed that the FPG,FINS,TG,TC, and LDL-C of the model group were significantly increased (P0.01), while the HDL-C was significantly decreased (P0.01), the H0MA-IR of the model group was also significantly increased (P0.01). The expression and phosphorylation of JNK protein in the model group were significantly higher than those in the control group (P < 0. 01). The results of electroacupuncture group and model group showed that FPG,FINS,TG,TC, and LDL-C in electroacupuncture group were significantly decreased (P < 0. 01), but HDL-C was significantly increased (P < 0. 01), and H0MA-IR in electroacupuncture group was also significantly decreased (P < 0. 01). In addition, the expression and phosphorylation of JNK protein in electroacupuncture group were significantly lower than those in control group (P < 0. 01). Conclusion: high fat diet can successfully induce insulin resistance in experimental rats and establish insulin resistance model. Electroacupuncture therapy can reduce the content of FPG,FINS,TG,TC and LDL-C and increase the content of HDL-C by acupuncture Sanyinjiao, Zusanli, Shenshu, Neiguan, invigorating spleen and kidney, dispelling blood stasis and removing phlegm, and improving the disorder state of metabolism of glucose and lipid. At the same time, the expression and phosphorylation of JNK protein in aortic endothelial cells were down-regulated, and insulin signal transduction pathway was restored, thus improving insulin resistance in model rats, and the insulin sensitivity of model rats was also enhanced.
【学位授予单位】：广州中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R245

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