化浊颗粒改善2型糖尿病大鼠胰岛素抵抗与PI-3K、GLUT4mRNA表达的影响

发布时间：2018-08-07 08:30

【摘要】：目的：观察化浊颗粒对2型糖尿病模型大鼠磷脂酰肌醇3激酶(phosphatidylinositol-3-kinase,PI-3K)、葡萄糖转运4(glucose transporter4,GLUT4)在骨骼肌组织中表达的影响。制备2型糖尿病大鼠模型，以中药方制成的化浊颗粒干预T2DM大鼠，，来探讨化浊颗粒是否可通过影响PI-3K及GLUT4的信号通路的激活，进而改善胰岛素抵抗，防治2型糖尿病的分子机制。验证解毒化浊法改善胰岛素抵抗的科学性和合理性。为开发中药胰岛素增敏剂提供实验依据，进而为中医药防治胰岛素抵抗奠定基础。方法：选取60只SPF级健康、雄性wistar大鼠（4月龄，200±20g），适应性喂养1周，予普通饲料喂养，从中随机抽出12只作为空白对照，余下48只作为造模组，予高脂饲料喂养4周，采用高脂饲料喂养联合小剂量链脲佐菌素（streptozotocin,STZ,30mg.kg-1,连续给药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，测定大鼠尾静脉空腹血糖及OGTT2小时血糖。如果72h后造模大鼠的FBG≥7.0mmol/L，OGTT（2hPG）≥11.1mmol/L，即为造模成功。将造模成功大鼠随机分为化浊颗粒组(2.025g/kg)，罗格列酮组(0.36mg/kg)，模型组(1ml/100g)，继续高脂饲料喂养，并与空白组(1ml/100g)一起予以相应药物灌胃治疗，其中模型组与空白组均予以生理盐水灌胃，观察大鼠一般状态，体重及摄食量，干预10w以后，予10%水合氯醛麻醉大鼠，检测大鼠FBG、OGTT（2hPG）、FINS水平，并应用反转录聚合酶链式反应(RT-PCR)方法检测各组大鼠骨骼肌组织PI-3K和GLUT4mRNA水平的表达。全部数据采用SPSS19.0软件进行统计学分析。结果 1．高脂饲料喂养组的体重较普通饲料组显著增加，差异有统计学意义（P0.05），高脂饲料组与普通饲料组大鼠的空腹血糖对比，无显著性差异（P0.05），但予小剂量STZ（30mg.kg-1）腹腔注射后，高脂饲料组血糖水平可见明显升高，与普通饲料组相比，差异有统计学意义(P0.05)；实验中共成模36只，成模率为：36/48=75%，提示成功复制T2DM动物模型。 2.药物干预10w后，罗格列酮组及模型组大鼠的体重同空白组相比均有显著性增加（P0.05），化浊颗粒组与模型组相比，体重低于模型组，差异有统计学意义（P0.05），提示：化浊颗粒对肥胖的T2DM大鼠体重的增长有一定的降低作用。 3.造模成功的大鼠与空白组对比，FBG、2hPG、FINS水平均有升高，差异有统计学意义（P0.05），药物干预10w后，化浊颗粒组与罗格列酮组均能降低大鼠FBG、2hPG及FINS水平，与模型组对比，差异具有统计学意义（P0.05）；化浊颗粒组与罗格列酮组之间对比，无显著性差异（P0.05）。 4.RT-PCR结果：化浊颗粒与罗格列酮干预的T2DM大鼠组，其骨骼肌组织PI-3K及GLUT4的表达水平均较模型组，显著增加（P0.05）。模型组较空白组PI-3K、GLUT4的表达水平呈低表达（P0.05）。结论 1.化浊颗粒通过提高胰岛素敏感性，达到改善T2DM胰岛素抵抗的作用； 2.化浊颗粒提高胰岛素敏感性的作用，与激活骨骼肌组织胰岛素信号传导通路中PI-3K和GLUT4mRNA的表达有关。
[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.
【学位授予单位】：甘肃中医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R259;R285.5

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