【摘要】：目的：觀察化濁顆粒對2型糖尿病模型大鼠磷脂酰肌醇3激酶(phosphatidylinositol-3-kinase,PI-3K)、葡萄糖轉運4(glucose transporter4,GLUT4)在骨骼肌組織中表達的影響。制備2型糖尿病大鼠模型，以中藥方制成的化濁顆粒干預T2DM大鼠，，來探討化濁顆粒是否可通過影響PI-3K及GLUT4的信號通路的激活，進而改善胰島素抵抗，防治2型糖尿病的分子機制。驗證解毒化濁法改善胰島素抵抗的科學性和合理性。為開發(fā)中藥胰島素增敏劑提供實驗依據，進而為中醫(yī)藥防治胰島素抵抗奠定基礎。 方法：選取60只SPF級健康、雄性wistar大鼠（4月齡，200±20g），適應性喂養(yǎng)1周，予普通飼料喂養(yǎng)，從中隨機抽出12只作為空白對照，余下48只作為造模組，予高脂飼料喂養(yǎng)4周，采用高脂飼料喂養(yǎng)聯(lián)合小劑量鏈脲佐菌素（streptozotocin,STZ,30mg.kg-1,連續(xù)給藥3天）腹腔注射的方法，建立2型糖尿病大鼠模型，72h后，禁食12h、不禁水，測定大鼠尾靜脈空腹血糖及OGTT2小時血糖。如果72h后造模大鼠的FBG≥7.0mmol/L，OGTT（2hPG）≥11.1mmol/L，即為造模成功。將造模成功大鼠隨機分為化濁顆粒組(2.025g/kg)，羅格列酮組(0.36mg/kg)，模型組(1ml/100g)，繼續(xù)高脂飼料喂養(yǎng)，并與空白組(1ml/100g)一起予以相應藥物灌胃治療，其中模型組與空白組均予以生理鹽水灌胃，觀察大鼠一般狀態(tài)，體重及攝食量，干預10w以后，予10%水合氯醛麻醉大鼠，檢測大鼠FBG、OGTT（2hPG）、FINS水平，并應用反轉錄聚合酶鏈式反應(RT-PCR)方法檢測各組大鼠骨骼肌組織PI-3K和GLUT4mRNA水平的表達。全部數據采用SPSS19.0軟件進行統(tǒng)計學分析。 結果 1．高脂飼料喂養(yǎng)組的體重較普通飼料組顯著增加，差異有統(tǒng)計學意義（P0.05），高脂飼料組與普通飼料組大鼠的空腹血糖對比，無顯著性差異（P0.05），但予小劑量STZ（30mg.kg-1）腹腔注射后，高脂飼料組血糖水平可見明顯升高，與普通飼料組相比，差異有統(tǒng)計學意義(P0.05)；實驗中共成模36只，成模率為：36/48=75%，提示成功復制T2DM動物模型。 2.藥物干預10w后，羅格列酮組及模型組大鼠的體重同空白組相比均有顯著性增加（P0.05），化濁顆粒組與模型組相比，體重低于模型組，差異有統(tǒng)計學意義（P0.05），提示：化濁顆粒對肥胖的T2DM大鼠體重的增長有一定的降低作用。 3.造模成功的大鼠與空白組對比，F(xiàn)BG、2hPG、FINS水平均有升高，差異有統(tǒng)計學意義（P0.05），藥物干預10w后，化濁顆粒組與羅格列酮組均能降低大鼠FBG、2hPG及FINS水平，與模型組對比，差異具有統(tǒng)計學意義（P0.05）；化濁顆粒組與羅格列酮組之間對比，無顯著性差異（P0.05）。 4.RT-PCR結果：化濁顆粒與羅格列酮干預的T2DM大鼠組，其骨骼肌組織PI-3K及GLUT4的表達水平均較模型組，顯著增加（P0.05）。模型組較空白組PI-3K、GLUT4的表達水平呈低表達（P0.05）。 結論 1.化濁顆粒通過提高胰島素敏感性，達到改善T2DM胰島素抵抗的作用； 2.化濁顆粒提高胰島素敏感性的作用，與激活骨骼肌組織胰島素信號傳導通路中PI-3K和GLUT4mRNA的表達有關。
[Abstract]:Objective: To observe the effect of Hua Zhuo Granule on the expression of phosphatidylinositol 3 kinase (phosphatidylinositol-3-kinase, PI-3K) and glucose transport 4 (glucose Transporter4, GLUT4) in the skeletal muscle of type 2 diabetic rats. To prepare the model of type 2 diabetic rats and to intervene the turbidity granules in T2DM rats. Is it possible to improve the molecular mechanism of type 2 diabetes by affecting the activation of PI-3K and GLUT4 signaling pathways and further improving insulin resistance and preventing the molecular mechanism of type 2 diabetes, and validates the scientific and rational effect of detoxification and turbidity method to improve insulin resistance.
Methods: 60 SPF grade health, male Wistar rats (4 month old, 200 + 20g) were fed for 1 weeks, and 12 were randomly selected as blank control. The remaining 48 were used as models and fed with high fat feed for 4 weeks. High fat diet was used to feed low dose streptozotocin (streptozotocin, STZ, 30mg.kg-1, continuous). After 3 days of administration, the rat model of type 2 diabetes was established by intraperitoneal injection. After 72h, 12h was fasted and water was forbidden. The fasting blood glucose and OGTT2 hour blood sugar of the rat tail vein were measured. If the FBG of the rats was greater than 7.0mmol/L, OGTT (2hPG) was more than 11.1mmol/L after 72h, the rats were randomly divided into the turbidity granule group (2.025g/kg) and Roger. The group (0.36mg/kg), the model group (1ml/100g), continued high fat diet and fed with the blank group (1ml/100g). The model group and the blank group were gavage with the normal saline, and observed the general state of the rats, the weight and the intake of food. After the intervention of 10W, 10% chloral chloral anaesthetized rats were given, and FBG, OGTT (2hPG) was detected in rats. The level of FINS and the expression of PI-3K and GLUT4mRNA in skeletal muscle tissues of each group were detected by reverse transcription polymerase chain reaction (RT-PCR). All data were statistically analyzed by SPSS19.0 software.
Result
1. the body weight of the high fat feed group was significantly higher than that of the ordinary diet group, the difference was statistically significant (P0.05). There was no significant difference in the fasting blood glucose of the high fat diet group and the normal diet group (P0.05), but the blood glucose level in the high fat diet group was obviously increased after the low dose STZ (30mg.kg-1) intraperitoneal injection, compared with the ordinary diet group. The difference was statistically significant (P0.05). There were 36 models in the experiment, and the rate of mold formation was 36/48=75%, indicating that the T2DM animal model was successfully replicated.
2. after 10W intervention, the weight of the rats in the rosiglitazone group and the model group were significantly higher than that in the blank group (P0.05). The body weight was lower than the model group, and the difference was statistically significant (P0.05). The results suggested that the granules of turbidity had a certain effect on the weight growth of obese T2DM rats.
3. compared with the blank group, the levels of FBG, 2hPG and FINS were higher than those in the blank group. The difference was statistically significant (P0.05). After the intervention of 10W, the level of FBG, 2hPG and FINS in the group of turbidity particles and rosiglitazone could reduce the level of FBG, 2hPG and FINS, compared with the model group, the difference had the significance of unified planning (P0.05), and the comparison between the turbidity particle group and the rosiglitazone group was compared with the model group. There was no significant difference (P0.05).
4.RT-PCR results: the expression level of PI-3K and GLUT4 in skeletal muscle tissue was significantly increased in T2DM rats treated with turbidity and rosiglitazone (P0.05). The expression level of GLUT4 in the model group was lower than that in the blank group (P0.05).
conclusion
1. Hua Hu granule can improve insulin resistance by enhancing insulin sensitivity in T2DM.
2. The effect of Huazhuo Granule on insulin sensitivity is related to the activation of PI-3K and GLUT4 mRNA expression in insulin signal transduction pathway of skeletal muscle tissue.
【學位授予單位】：甘肅中醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R259;R285.5

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