背景:糖尿病(DM)患者的肌肉健康常常被忽视,导致肌肉萎缩.DM中活性氧的增加可以降低抗氧化酶,例如超氧化物歧化酶-1(SOD-1)和-2(SOD-2),并抑制钙调磷酸酶(CN)和PGC-1α信号通路。已知绿原酸(CGA)是CN和PGC-1α的有效抗氧化剂和活化剂。本研究旨在确定CGA对SOD-1,SOD-2,CN和PGC-1αmRNA表达的影响,以抑制DM向肌肉萎缩的进展。
方法:本研究在解剖学,医学院,公共卫生,和护理,GadjahMada大学于7月20日开始,2020年。将24只雄性Wistar大鼠随机分为6组,每组4只。即,control,DM1.5个月(DM1.5),和DM2个月(DM2);和以三种不同剂量的CGA治疗的DM组,即CGA1(12.5毫克/千克体重),CGA2(25毫克/千克体重),和CGA3(50mg/kgBW)。对照组仅注射生理盐水,而链脲佐菌素腹腔注射诱导糖尿病模型。测量血糖水平两次(糖尿病诱导后一周和终止前)。收集比目鱼肌组织,采用RT-PCR方法分析SOD-1,SOD-2,CN和PGC-1α的mRNA表达。此外,对组织样本进行CN和苏木精-伊红(HE)免疫组织化学染色,在光学显微镜下进行形态学分析.
结果:与DM2组相比,CGA1组的SOD-1和SOD-2的mRNA表达相对较高。CGA1组CN的mRNA表达显著高于DM2组(p=0.008)。CGA1组PGC-1α的mRNA表达明显高于DM2组(p=0.025)。免疫组化染色显示CGA1组CN免疫阳性表达较其他组更为明显。苏木精-伊红染色显示CGA1组肌肉组织形态与对照组相似。
结论:绿原酸在12.5mg/kg体重的剂量下显示较低的血糖水平,与DM组相比,骨骼肌组织形态良好,SOD-1,SOD-2,CN和PGC-1α的mRNA表达更高。
Muscle health in diabetes mellitus (DM) is often neglected, which leads to muscle wasting. Increased reactive oxygen species in DM could decrease antioxidant enzymes such as superoxide dismutase-1 (SOD-1) and -2 (SOD-2) and inhibit calcineurin (CN) and PGC-1α signalling pathways. Chlorogenic acid (CGA) is known as a potent antioxidant and activators of CN and PGC-1α. This study aimed to determine the effect of CGA on mRNA expressions of SOD-1, SOD-2, CN and PGC-1α in inhibiting the progression of DM to muscle wasting.
This study was conducted at Department of Anatomy, Faculty of Medicine, Public Health, and Nursing, Universitas Gadjah Mada starting on July 20th, 2020. A total of 24 male Wistar rats were randomly divided into six groups (four rats per group), i.e., control, DM 1.5 months (DM1.5), and DM 2 months (DM2); and DM groups treated with CGA in three different doses, namely CGA1 (12.5 mg/kg BW), CGA2 (25 mg/kg BW), and CGA3 (50 mg/kg BW). Control group was only injected with normal saline, while diabetic model was induced by intraperitoneal injection of streptozotocin. Blood glucose levels were measured twice (one week after diabetic induction and before termination). The soleus muscle tissue was harvested to analyse the mRNA expressions of SOD-1, SOD- 2, CN and PGC-1α using RT-PCR. In addition, the tissue samples were stained with immunohistochemistry for CN and haematoxylin-eosin (HE) for morphologic analysis under light microscopy.
The mRNA expressions of SOD-1 and SOD-2 in the CGA1 group were relatively higher compared to the DM2 groups. The mRNA expression of CN in the CGA1 group was significantly higher compared to the DM2 group (p = 0.008). The mRNA expression of PGC-1α in the CGA1 group was significantly higher compared to the DM2 group (p = 0.025). Immunohistochemical staining showed that CNimmunopositive expression in the CGA1 group was more evident compared to the other groups. Haematoxylin-eosin staining showed that muscle tissue morphology in the CGA1 group was similar to that in the control group.
Chlorogenic acid at a dose of 12.5 mg/kg BW shows lower blood glucose level, good skeletal muscle tissue morphology and higher mRNA expressions of SOD-1, SOD-2, CN and PGC-1α compared to the DM groups.