電針對高脂喂養(yǎng)大鼠主動脈內皮細胞JNK蛋白的影響
發(fā)布時間：2018-10-24 20:26
【摘要】：目的:本次實驗在前期實驗研究的基礎上,將高脂飲食誘導的胰島素抵抗模型大鼠進行電針干預治療,觀察主動脈內皮細胞JNK蛋白的變化,探究并完善電針改善胰島素抵抗狀態(tài)的內在機制,同時也為臨床治療胰島素抵抗相關疾病提供參考。方法:在SPF級實驗室標準環(huán)境下,本次實驗研究設立3個組,分別為空白組、模型組和電針組,實驗動物選擇雄性SD大鼠24只,隨機分到3組,每組8只動物。空白組的實驗大鼠選用普通飼料進行喂養(yǎng),而模型組與電針組的實驗大鼠則采用高脂飼料進行喂養(yǎng)。喂養(yǎng)12周后,檢測三組實驗模型大鼠的空腹胰島素(FINS)和空腹血糖(FPG)數據并計算穩(wěn)態(tài)模型評價的胰島素抵抗指數(HOMA-IR),從而評價模型的建立是否成功。造模成功后,電針組的實驗大鼠選擇雙側內關穴、三陰交穴、足三里穴以及腎俞穴進行針刺干預治療,其中雙側足三里穴和三陰交穴采用電針療法進行干預,并繼續(xù)以高脂飼料喂養(yǎng)。電針治療選用疏密波(密波50 Hz,疏波2 Hz,頻率轉換10次/min),其強度以實驗大鼠后肢輕微抖動為度,20 min/次,1次/d,持續(xù)4w。而空白組與模型組的實驗大鼠則繼續(xù)分別予普通飼料和高脂飼料進行喂養(yǎng),僅采用與電針組相同的按壓固定時間,其余不做干預。4周后,檢測三組實驗大鼠的空腹胰島素(FINS)、空腹血糖(FPG)、總膽固醇(TC)、甘油三酯(TG)、高密度脂蛋白(HDL-C)、低密度脂蛋白(LDL-C)以及胰島素抵抗指數(H0MA-IR)的數據并計算穩(wěn)態(tài)模型評價的胰島素抵抗指數(H0MA-IR)。采用Western-blotting方法檢測主動脈內皮細胞JNK蛋白的表達及其磷酸化水平。結果:喂養(yǎng)12周后,電針組與模型組實驗大鼠FPG、FINS以及H0MA-IR相比于空白組有顯著升高(P＜0.0);電針治療4周后,比較空白組與模型組結果:模型組大鼠FPG、FINS、TG、TC、以及LDL-C均有顯著升高(P0.01),而HDL-C則明顯降低(P0.01);模型組大鼠H0MA-IR也有顯著升高(P0.01);此外,模型組大鼠的JNK蛋白的表達及其磷酸化水平比空白組大鼠也有顯著增加(P＜0.01)。比較電針組和模型組的結果:電針組大鼠FPG、FINS、TG、TC、以及LDL-C均有顯著降低(P＜0.01),而HDL-C則明顯升高(P＜0.01);電針組大鼠H0MA-IR也有顯著減低(P＜0.01)。此外,電針組大鼠的JNK蛋白的表達及其磷酸化水平比空白組大鼠也有顯著降低(P＜0.01)。結論:高脂飲食喂養(yǎng)大鼠能夠成功誘導實驗大鼠胰島素抵抗的狀態(tài),實現胰島素抵抗模型的建立。電針療法通過針刺三陰交、足三里、腎俞穴、內關,健脾補腎、祛瘀化痰通絡,降低大鼠體內FPG、FINS、TG、TC以及LDL-C的含量,并提高HDL-C的含量,從而改善機體內糖代謝和脂代謝的紊亂狀態(tài);與此同時,下調主動脈內皮細胞JNK蛋白的表達及其磷酸化水平,恢復胰島素信號傳導通路,從而改善模型大鼠胰島素抵抗狀態(tài),且模型大鼠對胰島素的敏感性也有增強。
[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.
【學位授予單位】：廣州中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R245
本文編號：2292460
[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.
【學位授予單位】：廣州中醫(yī)藥大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R245
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