Universitas Muhammadiyah Surabaya, Faculty of Health Sciences, Department of Physiotherapy, Surabaya, Indonesia.
Universitas Ahmad Dahlan, Faculty of Medicine, Department of Anatomy, Yogyakarta, Indonesia.
Med J Malaysia. 2024 Aug;79(Suppl 4):23-30.
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.
糖尿病（DM）患者的肌肉健康常常被忽视，这导致肌肉萎缩。DM 中活性氧的增加会降低超氧化物歧化酶-1（SOD-1）和 -2（SOD-2）等抗氧化酶，并抑制钙调神经磷酸酶（CN）和过氧化物酶体增殖物激活受体 γ 共激活因子 1α（PGC-1α）信号通路。绿原酸（CGA）是一种有效的抗氧化剂，可激活 CN 和 PGC-1α。本研究旨在确定 CGA 对 SOD-1、SOD-2、CN 和 PGC-1α mRNA 表达的影响，以抑制 DM 向肌肉萎缩的进展。
本研究于 2020 年 7 月 20 日在日惹加查马达大学医学、公共卫生和护理学院解剖系进行。总共 24 只雄性 Wistar 大鼠被随机分为六组（每组 4 只大鼠），即对照组、DM 1.5 个月（DM1.5）和 DM 2 个月（DM2）；和 DM 组分别用三种不同剂量的 CGA 治疗，即 CGA1（12.5mg/kgBW）、CGA2（25mg/kgBW）和 CGA3（50mg/kgBW）。对照组仅注射生理盐水，而糖尿病模型则通过腹腔注射链脲佐菌素诱导。在糖尿病诱导后一周和终止前两次测量血糖水平。使用 RT-PCR 分析比目鱼肌组织中 SOD-1、SOD-2、CN 和 PGC-1α 的 mRNA 表达。此外，还使用免疫组织化学法对 CN 进行染色，并用苏木精-伊红（HE）对光镜下的组织样本进行形态分析。
CGA1 组的 SOD-1 和 SOD-2 的 mRNA 表达相对较高。与 DM2 组相比，CGA1 组的 CNmRNA 表达明显升高（p=0.008）。与 DM2 组相比，CGA1 组的 PGC-1αmRNA 表达明显升高（p=0.025）。免疫组织化学染色显示，与其他组相比，CGA1 组的 CN 免疫阳性表达更为明显。苏木精-伊红染色显示，CGA1 组的肌肉组织形态与对照组相似。
与 DM 组相比，绿原酸 12.5mg/kgBW 可降低血糖水平，具有良好的骨骼肌组织形态，SOD-1、SOD-2、CN 和 PGC-1α 的 mRNA 表达更高。
