青蒿琥酯基于TGF-β/Smad通路减轻2型糖尿病大鼠脾脏纤维化的机制研究

Mechanism of artesunate attenuating spleen fibrosis in type 2 diabetic rats based on TGF-β/Smad pathway

探讨青蒿琥酯(artesunate,ART)对2型糖尿病(type 2 diabetes mellitus,T2DM)大鼠脾脏纤维化的防治作用,并明确其相关分子机制,为T2DM脾脏损伤防治提供实验依据。50只SD实验大鼠分为正常组和模型组,模型组高糖高脂喂养8周后,注射链脲佐菌素构建T2DM模型,再随机分为糖尿病组、二甲双胍治疗组、青蒿琥酯治疗组、联合治疗组,连续用药干预4周后取大鼠脾脏,称脾脏重量;HE、Masson染色观察脾脏病理变化;免疫组织化学法和Western blot检测脾脏组织中转化生长因子β1(transforming growth factor-β,TGF-β1)、重组人蛋白2(smad family member 2,Smad2)、重组人蛋白7(smad family member 7,Smad7)、Ⅰ型胶原(CollagenⅠ)表达;qRT-PCR检测脾脏组织中TGF-β1、Smad2、Smad7、CollagenⅠ的mRNA表达量。与正常组比较,糖尿病组大鼠脾脏指数下降,脾脏细胞排列紊乱,大量纤维化增生。与糖尿病组比较,各用药干预组的脾脏损伤和纤维化减轻,脾脏组织中TGF-β1、Smad2、CollagenⅠ蛋白和mRNA含量显著降低、Smad7明显上升。以上结果说明ART能够减轻T2DM大鼠脾脏损伤,抑制脾脏纤维化增生,其机制可能与抑制TGF-β/Smad信号通路有关。

To investigate the preventive and therapeutic effects of artesunate(ART)on splenic fibrosis in type 2 diabetes mellitus(T2DM)rats,and to reveal its related molecular mechanisms,providing a experimental basis for the prevention and treatment of splenic injury in T2DM.Fifty SD rats were randomly divided into control group and model group.After eight-week high glucose and high fat feeding in the model group,streptozotocin was injected to make a T2DM model,and then they were randomly divided into diabetes group,metformin intervention group,artesunate intervention group,metformin and artesunate intervention group.After four-week treatment,the spleen tissues of rats were harvested.The spleen was weighed;HE,Masson staining were used to observe the pathological changes of the spleen;Immunohistochemistry,Western blot were used to detect the expression of transforming growth factor-β(TGF-β1),SMAD family member 2(Smad2),SMAD family member 7(Smad7),and type I collagen in the spleen tissues;And qRT-PCR was used to detect the mRNA levels of TGF-β1,Smad2,Smad7,and collagen I in the spleen tissues.The spleen index decreased.The structure of spleen was damaged and disorganized.Compared with control group,there were fibrosis and hyperplasia in spleen observed in diabetes group.Compared with the diabetes group,the spleen injury and fibrosis were attenuated in drug intervention group.The expression and mRNA contents of TGF-β1,Smad2,and collagen I in the spleen tissues significantly decreased.The expression and mRNA content of Smad7 significantly increased.ART ameliorated splenic injury and inhibited splenic fibrosis in T2DM rats,and the mechanism of action may be related to the inhibition of TGF-β/Smad signaling pathway.