TLR2和TLR4在糖尿病动脉硬化大鼠模型中的表达

发布时间：2018-07-08 20:18

本文选题：TLR2 + TLR4　；参考：《广西医科大学》2012年硕士论文

【摘要】：背景与目的：近年来,国内糖尿病(diabetes mellitus, DM)患病率迅速升高。慢性大血管病变是糖尿病损害一个主要表现,糖尿病动脉硬化(diabetic arteriosclerosis, DAS)是导致糖尿病患者死亡的重要原因之一。目前关于糖尿病及动脉粥样硬化(athemsclemsis, AS)的发病机制还不是很明确,建立动物模型是实验研究的基础。“损伤反应学说”认为慢性炎症过程和血脂代谢紊乱是AS的两个主要特点。Toll样受体(toll-like receptors, TLR)是近年来发现的天然免疫受体,它们可以通过广泛地特异性地识别病原微生物,偶联信号转导途径,并激活天然免疫细胞,最终导致一系列的免疫炎症反应,有可能在DAS过程中发挥了重要的作用。本课题通过检测糖尿病动脉粥样硬化大鼠模型中TLR2、TLR4的表达,探讨TLR2及TLR4在糖尿病动脉粥样硬化中起到的作用。材料与方法：60只雄性SD大鼠随机分为两组,30只作为正常对照组,其余作为实验对照组。用腹腔注射链脲佐菌素(streptozotocin, STZ)建立糖尿病动脉硬化大鼠模型。高糖高脂喂食后检测血糖、总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白(HDL)、低密度脂蛋白(LDL);用逆转录聚合酶链反应(retrovirus polymerase chain reaction, RT-PCR)方法检测血液中TLR2及TLR4mRNA表达情况,并用凝胶扫描仪进行电泳条带强度分析；用免疫组织化学(immunohistochemical method) SP法检测TLR2及TLR4在动脉组织中的蛋白表达情况；观察动脉并取病变处,送广西医科大学试验中心制作HE切片,光镜下观察动脉壁改变。结果： 1.动物模型制造连续3次STZ注射完毕后,与对照组相比,模型组大鼠血糖明显增高(P0.01)；高糖高脂喂食8周之后,模型组大鼠血脂四项,明显增高(P0.01),造模成功率为93.33%(28/30)。 2. RT-PCR检测28只糖尿病动脉硬化大鼠血浆中TLR2mRNA的阳性率为100%(28/28),相对应的正常组大鼠血浆阳性率为40%(12/30),两组比较均有显著差异(P0.01)。检测28只糖尿病动脉硬化大鼠血浆中TLR4mRNA的阳性率为100%(28/28),相对应的正常组大鼠血浆阳性率为23.33%(7/30),两组比较均有显著差异(P0.01)。 3.免疫组化检测28只糖尿病动脉硬化大鼠血浆中TLR2蛋白的阳性率为89.28%(25/28),对照组大鼠血浆阳性率为13.33%(4/30),两组比较均有显著差异(P0.01)。检测28只糖尿病动脉硬化大鼠血浆中TLR4蛋白的阳性率为92.86%(26/28),对照组大鼠血浆阳性率为6.67%(2/30),两绀比较均有显著差异(P0.01)。 4.经Spearmann、Pearson等级相关分析TLR2和TLR4的mRNA和蛋白表达水平呈正相关关系(P0.01)。 5.病理切片主动脉肉眼可见动脉管肥大,镜下观具有糖尿病动脉硬化的早期改变,如纤维组织增生、有核细胞增多,表层及中膜层结构变疏松。结论：研究结果显示,TLR2和TLR4在糖尿病动脉硬化大鼠主动脉和血浆中高表达,这种表达与高血糖及血脂代谢紊乱有关。对两者的联合检测尤其是早期检测有助于提早发现糖尿病动脉硬化改变,为以后的临床治疗提供一定的帮助。
[Abstract]:BACKGROUND & OBJECTIVE : In recent years , the prevalence of diabetes mellitus ( DM ) is increasing rapidly . The pathogenesis of diabetes and atherosclerosis is one of the most important causes of diabetes . Toll - like receptors ( TLR ) are two main characteristics of AS . Toll - like receptors ( TLR ) are two main characteristics of AS . Toll - like receptors ( TLR ) have played an important role in the pathogenesis of diabetes mellitus . Toll - like receptors ( TLR ) can play an important role in the process of DAS .

Materials and Methods : Sixty male SD rats were randomly divided into two groups , 30 were used as control group , and the rest were used as experimental control group . Blood glucose , total cholesterol ( TC ) , triglyceride ( TG ) , high density lipoprotein ( HDL ) and low density lipoprotein ( LDL ) were detected after high glucose and high fat feeding . The expression of TLR2 and TLR4 mRNA in blood was detected by reverse transcription polymerase chain reaction ( RT - PCR ) .
Immunohistochemical method was used to detect the expression of TLR2 and TLR2 in arterial tissue .
The artery was observed and the lesion was taken , HE section was produced at the experimental center of Guangxi Medical University , and the changes of arterial wall were observed under light microscope .

Results :

1 . The blood glucose of the model group was significantly higher than that of the control group ( P0.01 ) .
After 8 weeks of high glucose and high fat diet , the blood lipids in the model group increased significantly ( P0.01 ) , and the success rate of the model group was 93.33 % ( 28 / 30 ) .

2 . The positive rate of TLR2 mRNA in plasma of 28 diabetic arteriosclerosis rats was 100 % ( 28 / 28 ) and 40 % ( 12 / 30 ) respectively . The positive rate of TLR4 mRNA was 100 % ( 28 / 28 ) in 28 diabetic arteriosclerosis rats .

3 . The positive rate of TLR2 protein in plasma of 28 diabetic arteriosclerosis rats was 89.28 % ( 25 / 28 ) and 13.33 % ( 4 / 30 ) in control group .

4 . There was positive correlation between TLR2 and TLR2 mRNA and protein expression levels ( P0.01 ) .

5 . The vascular hypertrophy of the aorta is seen in the pathological section of the aorta , and the view of the mirror has the early change of the diabetic arteriosclerosis , such as fibrous tissue hyperplasia , the increase of nucleated cells , the change of the superficial layer and the middle membrane layer .

Conclusion : The results show that TLR2 and TLR2 are highly expressed in aorta and plasma of diabetic arteriosclerosis rats . This expression is related to hyperglycemia and lipid metabolism disorder . Combined detection of both is helpful for early detection of diabetic arteriosclerosis and provides some help for later clinical treatment .
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R587.1;R-332

【参考文献】