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目的通过链脲佐菌素(STZ)腹腔注射,建立胰岛素抵抗大鼠模型,探讨胰岛素抵抗大鼠Klotho的表达变化与死亡率的关系。方法出生当日Wistar大鼠80只,随机选取20只为正常对照组(CN组,n=20),其余60只腹腔注射STZ复制胰岛素抵抗模型。选取40只设立胰岛素抵抗组(IR组,n=20)、氨基胍联合川芎嗪治疗组(TMP+AG组,n=20)。分别在造模8、24、32周短尾取空腹血检测血糖(PG)、糖化血清蛋白(GSP)。造模32周后,取其肾脏实时荧光定量RT-PCR,检测Klotho mRNA的表达,Western blot法和免疫组织化学染色方法检测Klotho蛋白表达。结果与胰岛素抵抗组相比,氨基胍联合川芎嗪治疗组大鼠肾脏中Klotho表达明显上升,死亡率明显降低,差异有统计学意义(P<0.05);与正常对照组相比,胰岛素抵抗组Klotho表达明显降低,死亡率明显升高,差异有统计学意义(P<0.05)。结论胰岛素抵抗大鼠肾脏Klotho的表达水平与死亡率相关。
Objective To establish a rat model of insulin resistance by intraperitoneal injection of streptozotocin (STZ) and investigate the relationship between Klotho expression and mortality in insulin resistant rats. Methods 80 Wistar rats were born on the day of birth. Twenty rabbits were randomly selected as normal control group (CN group, n = 20). The remaining 60 rabbits were injected intraperitoneally with STZ replication insulin resistance model. Forty insulin resistance group (IR group, n = 20) and aminoguanidine combined with ligustrazine group (TMP + AG group, n = 20) were selected. Blood glucose (PG) and glycosylated serum protein (GSP) were determined by fasting blood samples at 8, 24 and 32 weeks after modeling. After 32 weeks of modeling, real-time fluorescent quantitative RT-PCR of kidney was used to detect the expression of Klotho mRNA. Klotho protein expression was detected by Western blot and immunohistochemical staining. Results Compared with insulin resistance group, the expression of Klotho in the kidney of aminoguanidine plus ligustrazine group was significantly increased, the mortality rate was significantly lower (P <0.05); compared with the normal control group, insulin resistance group Klotho expression was significantly reduced, the mortality was significantly increased, the difference was statistically significant (P <0.05). Conclusion The expression of Klotho in the kidney of insulin resistance rats is related to the mortality rate.