Al-Shaqha Waleed M, Khan Mohsin, Salam Nasir, Azzi Arezki, Chaudhary Anis Ahmad
College of Medicine, Al-Imam Mohammad Ibn Saud Islamic University (IMSIU), Riyadh, 13317-7544, Kingdom of Saudi Arabia (KSA).
Department of Energy and Environmental sciences, Chaudhary Devi Lal University, Sirsa, Haryana, 125055, India.
BMC Complement Altern Med. 2015 Oct 21;15:379. doi: 10.1186/s12906-015-0899-6.
Catharanthus roseus is an important Ayurvedic medication in traditional medicine. It is potentially used in countries like India, South Africa, China and Malaysia for the healing of diabetes mellitus. Although, the molecular mechanisms behind this effect are yet to be exclusively explored. Due to the great antidiabetic and hyperlipidemic potential of c. roseus, we hypothesized that the insulin mimetic effect of ethanolic extract of c. roseus might add to glucose uptake through improvement in the expression of genes of the glucose transporter (GLUT) family messenger RNA (mRNA) in liver.
STZ-induced diabetic rats treated by ethanolic extract of c. roseus 100 mg/kg and 200 mg/kg; and one group treated with Metformin (100 mg/kg). After final administration of treatment of 4 weeks, blood samples were collected under fasting conditions, and the body weights (BWs) were measured. Total RNA from liver was extracted with the Qiagen RNEasy Micro kit (GERMANY) as described in the manufacturer's instructions. First-strand complementary DNA (cDNA) was synthesized at 40 °C by priming with oligo-dT12-18 (Invitrogen, USA) and using Super ScriptII reverse transcriptase according to the protocol provided by the manufacturer (Invitrogen, USA). Real-time polymerase chain reaction (PCR) amplifications for GLUT-4 (gene ID: 25139) were conducted using Light-Cycler 480 (Roche, USA) with the SyBr® I nucleic acid stain (Invitrogen, USA) according to the manufacturer's instructions. Polymerase chain reaction products of β-actin primer gene were used as an internal standard.
The proposed study was framed to look at the antidiabetic efficacy of ethanolic extract of c. roseus and an expression of GLUT-2 and GLUT-4 gene in streptozotocin induced diabetic wistar rats. The doses were administered orally at a rate of 100 and 200 mg/kg and detrain the glucose transport system in liver for 4 weeks. The observed results showed a good positive correlation between intracellular calcium and insulin release levels in isolated islets of Langerhans. The supplementation of ethanolic extract of c. roseus significantly amplified the expression of GLUT gene mRNA by Real Time PCR in liver of diabetic rats.
We conclude that the observed antidiabetic effect of c. roseus on STZ induced diabetes was a result of complex mechanisms of GLUT gene mRNA expression. The findings are very encouraging and greatly advocate its candidature for the design of a novel herbal drug to cure deadly diabetes.
长春花是传统医学中一种重要的阿育吠陀药物。在印度、南非、中国和马来西亚等国家,它被潜在地用于治疗糖尿病。尽管如此,这种作用背后的分子机制尚未得到专门研究。由于长春花具有巨大的抗糖尿病和降血脂潜力,我们推测长春花乙醇提取物的胰岛素模拟作用可能通过改善肝脏中葡萄糖转运蛋白(GLUT)家族信使核糖核酸(mRNA)基因的表达来增加葡萄糖摄取。
用100mg/kg和200mg/kg的长春花乙醇提取物处理链脲佐菌素诱导的糖尿病大鼠;另一组用二甲双胍(100mg/kg)处理。在最后一次给药4周后,在禁食条件下采集血样,并测量体重(BW)。按照制造商说明书,使用Qiagen RNEasy Micro试剂盒(德国)从肝脏中提取总RNA。按照制造商(美国Invitrogen)提供的方案进行操作,以oligo-dT12-18(美国Invitrogen)为引物,在40°C下使用Super ScriptII逆转录酶合成第一链互补DNA(cDNA)。使用Light-Cycler 480(美国罗氏)和SyBr® I核酸染料(美国Invitrogen),按照制造商说明书对GLUT-4(基因ID:25139)进行实时聚合酶链反应(PCR)扩增。β-肌动蛋白引物基因的聚合酶链反应产物用作内标。
本研究旨在观察长春花乙醇提取物的抗糖尿病疗效以及链脲佐菌素诱导的糖尿病Wistar大鼠中GLUT-2和GLUT-4基因的表达。以100和200mg/kg的剂量口服给药,并在4周内抑制肝脏中的葡萄糖转运系统。观察结果显示,在分离的胰岛中,细胞内钙与胰岛素释放水平之间存在良好的正相关。补充长春花乙醇提取物通过实时PCR显著增强了糖尿病大鼠肝脏中GLUT基因mRNA的表达。
我们得出结论,长春花对链脲佐菌素诱导的糖尿病的抗糖尿病作用是GLUT基因mRNA表达复杂机制的结果。这些发现非常令人鼓舞,并大力支持其作为一种新型草药药物治疗致命糖尿病的候选药物。