Department of Pharmacy, Jashore University of Science and Technology, Jashore, 7408, Bangladesh; School of Biomedical Sciences and Graham Centre for Agricultural Innovation, Charles Sturt University, Boorooma St, Wagga Wagga, NSW, Australia.
School of Biomedical Sciences and Graham Centre for Agricultural Innovation, Charles Sturt University, Boorooma St, Wagga Wagga, NSW, Australia.
J Ethnopharmacol. 2021 Apr 24;270:113781. doi: 10.1016/j.jep.2021.113781. Epub 2021 Jan 7.
Phragmites karka (Retz.) of family Poaceae is a pristine tropical plant that is well known to the local healers for ailments of diabetes, fever, diarrhea and CNS depression but lacks the scientific evidence behind its traditional usage. Hence, we explicated this plant to find the scientific basis of its traditional utilization.
The current study aims to find out the antidiabetic potential, toxicity after oral administration and in vitro cytotoxic activity of aerial parts of the plant on HeLa cells.
The plant was extracted with methanol by maceration and the crude extract was then subjected to solvent partitioning with modified Kupchan method for preparing several fractions. Phytochemical screening and total phenolic content of the plant was first determined through established procedures. Acute toxicity of the plant was studied by orally administering a single high dose (5000 mg/kg) of drug. Cytotoxicity of the methanolic plant extract was determined by measuring the percentage of cell viability on human cervical cancer cell lines, HeLa. In vitro antidiabetic activity was determined through iodine starch and DNSA (3,5-dinitrosalicylic acid) method of α-amylase inhibition. Finally, in vivo oral glucose tolerance test and alloxan induced antidiabetic activity test was performed at 150 and 300 mg/kg body weight doses of plant extract to confirm the in vivo antidiabetic activity.
No mortality was demonstrated by Phragmites karka in the acute toxicity test. However, signs of cellular toxicity was observed and histopathological studies on major organs exhibited necrosis in liver and kidney. In vitro cytotoxicity assay revealed the death of HeLa cells by DCM (dichloromethane) and n-hexane fractions of plant extract at 100 and 10 μg/mL concentrations. The IC value of the fractions were later evaluated by MTT assay (316.1 and 96.7 μg/mL for n-hexane and DCM fractions, respectively). In the iodine starch and DNSA method of α-amylase enzyme inhibitory activity test, substantial inhibition of enzyme was observed with the IC values of 2.05 and 2.08 mg/mL, respectively. In the in vivo antidiabetic activity test, considerable reduction in blood glucose level of diabetic mice was detected in both oral glucose tolerance test and alloxan induced antidiabetic activity test. In addition, the microscopic examination of pancreas showed noticeable recovery of pancreatic β cells and the blood lipid profile analysis represented a significant (p < 0.05) reduction of total cholesterol, LDL (low density lipoprotein) and triglyceride level in plant extract treated mice.
Results of this study reveals that the Phragmites karka extract is toxic at cellular level after oral administration and cytotoxic when tested on HeLa cells. The plant also evidenced hypoglycemic property, possibly through the inhibition of α-amylase enzyme and recovered the pancreatic beta cells along with the improvement of lipid profile of diabetic mice. However, robust studies on this plant is required to isolate the bioactive compounds, elucidate structures and evaluate their mechanism of actions in support of our findings.
Toxicology and Safety, Quality Traditional Medicine.
禾本科植物菰（Retz.）是一种原始的热带植物，当地的治疗师熟知它可以治疗糖尿病、发烧、腹泻和中枢神经系统抑制等疾病，但缺乏其传统用途背后的科学证据。因此，我们对这种植物进行了阐述，以寻找其传统用途的科学依据。
本研究旨在发现该植物地上部分的抗糖尿病潜力、口服后的毒性和体外对 HeLa 细胞的细胞毒性。
采用浸渍法用甲醇提取植物，然后用改良的 Kupchan 法进行溶剂萃取，制备几种馏分。首先通过既定程序确定植物的植物化学筛选和总酚含量。通过口服给予单一大剂量（5000mg/kg）药物来研究植物的急性毒性。通过测量人宫颈癌细胞系 HeLa 细胞的存活率来确定甲醇植物提取物的细胞毒性。通过碘淀粉和 DNSA（3,5-二硝基水杨酸）法测定α-淀粉酶抑制作用，测定体外抗糖尿病活性。最后，在 150 和 300mg/kg 体重剂量的植物提取物进行体内口服葡萄糖耐量试验和链脲佐菌素诱导的抗糖尿病活性试验，以确认体内抗糖尿病活性。
菰在急性毒性试验中没有表现出死亡。然而，观察到细胞毒性的迹象，对主要器官的组织病理学研究显示肝脏和肾脏坏死。体外细胞毒性试验显示植物提取物的二氯甲烷（DCM）和正己烷馏分在 100 和 10μg/mL 浓度下杀死 HeLa 细胞。随后通过 MTT 测定法评估了馏分的 IC 值（正己烷和 DCM 馏分分别为 316.1 和 96.7μg/mL）。在碘淀粉和 DNSA 法测定α-淀粉酶酶抑制活性试验中，观察到酶的显著抑制，IC 值分别为 2.05 和 2.08mg/mL。在体内抗糖尿病活性试验中，在口服葡萄糖耐量试验和链脲佐菌素诱导的抗糖尿病活性试验中，糖尿病小鼠的血糖水平均显著降低。此外，胰腺的显微镜检查显示胰岛β细胞有明显恢复，血液脂质谱分析显示植物提取物治疗组的总胆固醇、低密度脂蛋白（LDL）和甘油三酯水平显著降低（p<0.05）。
本研究结果表明，菰提取物口服后在细胞水平上具有毒性，在 HeLa 细胞上具有细胞毒性。该植物还表现出降血糖作用，可能是通过抑制α-淀粉酶酶，恢复胰岛β细胞，并改善糖尿病小鼠的脂质谱。然而，需要对这种植物进行更深入的研究，以分离出生物活性化合物，阐明其结构，并评估其作用机制，以支持我们的研究结果。
毒理学与安全，传统药物质量。
