青蒿琥酯通过PI3K/GSK-3β通路对1型糖尿病小鼠胰岛素抵抗的改善作用研究

Effect of artesunate on insulin resistance in type 1 diabetic mouse through PI3K/GSK-3β pathway

关键词:  1型糖尿病  青蒿琥酯  胰岛素抵抗  PI3K/GSK3β信号通路

Key words:  type 1 diabetes  artesunate  insulin resistance  PI3K/GSK3β signaling pathway

摘要:

[目的] 观察青蒿琥酯对1型糖尿病小鼠胰岛素抵抗的影响，以及探究其作用机制。[方法] 取Balb/c小鼠腹腔注射200 mg/kg STZ建立1型糖尿病小鼠模型，随机分为模型组、青蒿琥酯组和胰岛素组。另随机选10只Balb/c小鼠为健康对照组，腹腔注射等量的生理盐水。建模成功后，青蒿琥酯组灌胃100 mg/kg青蒿琥酯，健康对照组和模型组灌胃等量的生理盐水，每日1次，共4周，胰岛素组皮下注射4 U/kg甘精胰岛素注射液，每日1次，连续5 d。检测各组小鼠空腹血糖（FBG）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）、肝脏三酰甘油（TG）、丙二醛（MDA）、血清游离脂肪酸（FFA）；苏木精-伊红（HE）染色观察小鼠胰岛病理学；蛋白免疫印迹法（Western blot）检测各组兔抗鼠磷脂酰肌醇3-激酶（PI3K）、磷酸化磷脂酰肌醇3-激酶（p-PI3K）、蛋白激酶B（Akt）、磷酸化蛋白激酶B（p-Akt）、糖原合成酶激酶-3（GSK3β）、磷酸化糖原合成酶激酶-3（p-GSK3β）和谷氨酰胺合成酶（GS）蛋白表达水平。[结果] 与健康对照组比较，模型组的FBG、FINS、HOMA-IR、TG、MDA和FFA含量显著增加（P＜0.05），p-PI3K/PI3K和p-Akt/Akt的蛋白比值以及GS蛋白表达量显著降低（P＜0.05），p-GSK3β/GSK3β蛋白表达量比值显著增加（P＜0.05）。与模型组比较，青蒿琥酯组、胰岛素组的FBG、FINS、HOMA-IR、TG、MDA和FFA含量显著降低（P＜0.05），p-PI3K/PI3K和p-Akt/Akt的蛋白比值以及GS蛋白表达量显著增加（P＜0.05），p-GSK3β/GSK3β蛋白表达量比值显著降低（P＜0.05）。[结论] 青蒿琥酯可显著降低1型糖尿病小鼠胰岛素抵抗，降低葡萄糖含量，其作用机制可能抑制PI3K/GSK3β信号通路传导有关。

Abstract:

[Objective] To observe the effects of artesunate on insulin resistance in type 1 diabetic mice,and to explore its mechanism of action. [Methods] Balb/c mice were injected intraperitoneally with 200 mg/kg STZ to establish a type 1 diabetic mouse model,and they were randomly divided into model group,artesunate group and insulin group. In addition,10 Balb/c mice were randomly selected as the healthy control group,and the same amount of normal saline was injected intraperitoneally. After successful modeling,the artesunate group was intragastrically administered with 100 mg/kg artesunate,and the healthy control group and model group were intragastrically administered with the same amount of normal saline once a day for 4 weeks,in the insulin group,4 U/kg insulin glargine injection was subcutaneously injected once a day for 5 consecutive days. Fasting blood glucose (FBG),fasting insulin (FINS),insulin resistance index (HOMA-IR),liver triglycerides (TG),malondialdehyde (MDA),and serum free fatty acids (FFA) were detected in each group of mice. HE staining was used to observe the pathology of mouse pancreatic islets. The expression levels of PI3K,p-PI3K,Akt,p-Akt,GSK3β,p- GSK3β and GS protein in each group were detected by Western blot. [Results] Compared with the healthy control group,the content of FBG,FINS,HOMA-IR,TG,MDA and FFA in the model group was increased significantly (P<0.05),the protein ratio of p-PI3K/PI3K and p-Akt/Akt and the expression of GS protein was decreased significantly (P<0.05),the expression ratio of p-GSK3β/GSK3β protein was significantly increased (P＜0.05). Compared with the model group,the contents of FBG,FINS,HOMA-IR,TG,MDA and FFA in the artesunate group and the insulin group were significantly reduced (P<0.05),the protein ratio of p-PI3K/PI3K and the expression level of p-Akt/Akt and GS protein were increased significantly (P<0.05),and the ratio of p-GSK3β/GSK3β protein expression level wass decreased significantly (P<0.05). [Conclusion] Artesunate can significantly reduce insulin resistance and glucose content in type 1 diabetic mice. Its mechanism of action maybe related to the inhibition of PI3K/GSK3β signaling pathway.