【作者】 余静；

【导师】 邱明才；

【作者基本信息】 天津医科大学 ， 内科学， 2010， 硕士

【摘要】 目的：以正常大鼠及无干预治疗的STZ诱导的DM大鼠作为对照,观察不同剂量Piog对STZ诱导的DM大鼠十二指肠的病理组织学变化,IL-10和TNF-α的表达,以及T淋巴细胞数目和比值的影响,并与免疫调节剂环孢菌素A(CsA)及胰岛素干预的大鼠相作比较,了解Piog是否通过影响CD4+/CD8+T淋巴细胞比值,TNF-α与IL-10等细胞因子的表达,来预防和控制糖尿病大鼠十二指肠的器官损伤,为临床上应用Piog控制DM及其并发症提供理论依据。方法：健康雄性SD大鼠96只,随机分为Piog小剂量干预组、Piog小剂量加INS干预组、Piog大剂量干预组、Cs A干预组、Cs A加INS干预组、INS干预组、DM无干预组和CON组等8组。除CON组外,其余均由STZ诱导造成DM模型。造模前及成模后每周检测大鼠体重,每月测定血糖,共饲养16周后处死。处死前取血清和尿检测生化指标。对各组大鼠十二指肠,采用HE染色、AB-PAS染色法观察十二指肠杯状细胞；免疫组织化学法检测各组大鼠十二指肠肌层Cajal间质细胞的表达,CD4+、CD8+T淋巴细胞的表达及TNF-α与IL-10的表达水平。结果：1.成功建立了STZ诱导的DM大鼠模型；2. Piog能明显降低血脂和改善肾功能；3.DM组大鼠十二指肠均有不同程度的粘膜固有层充血,水肿,绒毛损伤,各PIO或CsA治疗组大鼠十二指肠病变均有不同程度减轻；4.DM组大鼠十二指肠粘膜层杯状细胞数目明显低于CON组,各PIO或CsA治疗组大鼠十二指肠粘膜层杯状细胞数目均有不同程度增加；5.DM组大鼠十二指肠肌层Cajal间质细胞的数量减少,各PIO或CsA治疗组大鼠十二指肠粘膜层杯状细胞数目均有不同程度增加；6.DM组大鼠十二指肠CD4+T淋巴细胞数量增加,CD8+T淋巴细胞数量减少,且CD4+/CD8+T淋巴细胞比值增加；各PIO或CsA治疗组CD4+T淋巴细胞数量减少,CD8+T淋巴细胞数量增加,且CD4+/CD8+T淋巴细胞比值降低。7.DM组大鼠十二指肠抑炎因子IL-10的量减少,致炎因子TNF-α的量增加；各PIO或CsA治疗组抑炎因子IL-10的量增加,致炎因子TNF-α的量减少。结论：Piog除有INS增敏和降糖作用外,Piog能明显降低血脂和改善肾功能；糖尿病大鼠十二指肠粘膜T淋巴细胞亚群的变化和细胞因子IL-10,TNF-α的异常表达提示免疫机制和炎症反应参与了十二指肠的病理组织学损伤,引起十二指肠粘膜层杯状细胞,肌层Cajal间质细胞数目减少,导致糖尿病大鼠十二指肠动力障碍。各PIO或CsA治疗能改善十二指肠粘膜T淋巴细胞亚群的变化和细胞因子的异常表达,减轻了器官的炎症反应,增加杯状细胞和Cajal间质细胞数目。因此,Piog不但可作为一种降糖药物,而且可作为一种免疫调节剂,用于预防和控制DM的器官免疫损伤,减少功能障碍的发生。更多还原

【Abstract】 Objective:To investigate the effection of different dosege of the Piog on the function, pathology of the duodenum of the STZ-induced DM rats, as well as the the expression of the IL-10,TNF-aand the number and ratio of T cells on the same organs. And compare its effection with the immunomodulator cyclosporin A (CsA) and the insulin (INS). To find whether Piog has the effection to prevent and ameliorate the duodenum injury of the DM which through the immunizing mechanisms that mediated by the IL-10, TNF-aand ratio between CD4+ and CD8+ T lymphocytes. And provide the theoretical evidence for the usage of the Piog in the clinical practice.Materials and methods:Ninety-six healthy male SD rats were randomly divided into 7 DM groups and 1 healthy control group. According to the interventing mode, the DM groups were made by intravenous administration of STZ and then were divided into low dose Piog group, low dose Piog plus INS group, large dose Piog group, Cs A group, Cs A plus INS group, INS group, and DM group, respectively. The weight and blood glucose of each rat were investigated by routine test each month. After feeding 16 weeks, all rats were sacrificed. The serum was seperated and the urine was obtained to test the hepatic, renal function, triglyceride, insulin level and the urine albumin. The hematoxylin and eosin Stain, AB-PAS Stain were carried out to observe the goblet cells of the duodenum of the rats in different groups. On the same organs, the immunohistochemistry were carried out to detect the number of interstital cells of Cajal, CD4+ and CD8+T cells,deposition of the IL-10 antigen, and TNF-aantigen.Results:1. The STZ-induced DM rat model were successfully established; 2. Piog can significantly lower blood triglyceride and improving kidney function; 3. Congestion,edema and damage to the villus were found in the duodenum of DM group, Compared with the normal and DM control group,the duodenum of the large dose Piog interventing group rat and the CsA interventing DM groups are much better. 4. The number of goblet cells in the duodenum of DM group decreased,while compared with the DM group, Piog interventing group rat and the CsA interventing DM groups, The number of goblet cells in the duodenum increased to some extent.5. The number of Cajal cells in the duodenum of DM group decreased, while compared with the DM group,the Piog interventing group rat and the CsA interventing DM groups, The number of Cajal cells in the duodenum increased to some extent.6. The number of CD4+T cells in the duodenum of DM group increased, while The number of CD8+T cells decreased,the ratio between CD4+and CD8+T lymphocytes increased, compared with the DM group,the Piog interventing group rat and the CsA interventing DM groups, The number of CD4+T cells in the duodenum of DM group decreased, while The number of CD8+T cells increased,the ratio between CD4+ and CD8+ T lymphocytes decreased.7.Decreaed IL-10 and incresed TNF-a were found in DM group, while in the Piog interventing group rat and the CsA interventing DM groups, increaed IL-10 and decresed TNF-a were found.Conclusions:Apart from its insulin sensitization effect, Piog could also lower the blood triglyceride significantly and ameliorate the renal function of the DM rats. Diabetic rat duodenum changes in T lymphocyte subsets and inflammatory cytokines IL-10, TNF-a suggest abnormal expression of the immune system and inflammatory responses involved in the duodenum of pathological tissue damage, reduce of the number mucosal layer of goblet cells, muscularis of interstitial cells of Cajal, resulting in motor dysfunction of diabetic rat duodenum. The PIO, or CsA treatment can improve the duodenal mucosa T lymphocyte subsets and abnormal expression of cytokines, reduced organ inflammation, increased goblet cells and the number of interstitial cells of Cajal.Therefore, Piog is not only a medicine of INS sensitizer, but also is an immunomodulator for the preventing multi-organ immune damage of the DM.更多还原