Zahra Syeda Saniya, Ahmed Madiha, Qasim Muhammad, Gul Bilquees, Zia Muhammad, Mirza Bushra, Haq Ihsan-Ul
Department of Pharmacy, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan.
Institute of Sustainable Halophytes Utilization, University of Karachi, Karachi, 75270, Pakistan.
BMC Complement Altern Med. 2017 Sep 5;17(1):443. doi: 10.1186/s12906-017-1951-5.
The concept of botanical therapeutics has revitalized due to wide importance of plant derived pharmaceuticals. Therefore, the ameliorative characteristics of Ajuga bracteosa were studied.
Total phenolic content, flavonoid content, antioxidant capacity, reducing power and free-radical scavenging activity were determined colorimetrically. Specific polyphenols were quantified by RP-HPLC analysis. Preliminary cytotoxicity was tested using brine shrimp lethality assay while antiproliferative activity against THP-1 and Hep-G2 cell lines was determined by MTT and SRB protocols respectively. Antileishmanial potential was assessed via MTT colorimetric method. To investigate antidiabetic prospect, α-amylase inhibition assay was adopted whereas disc diffusion method was used to detect likely protein kinase inhibitory, antibacterial and antifungal activities.
Among fifteen different extracts, maximum total phenolic content (10.75 ± 0.70 μg GAE/mg DW), total reducing power (23.90 ± 0.70 μg AAE/mg DW) and total antioxidant capacity (11.30 ± 0.80 μg AAE/mg DW) were exhibited by methanol extract with superlative percent extract recovery (17.50 ± 0.80% w/w). Chloroform-methanol extract demonstrated maximum flavonoid content (4.10 ± 0.40 μg QE/mg DW) and ethanol extract exhibited greatest radical scavenging activity (IC 14.40 ± 0.20 μg/ml). RP-HPLC based quantification confirmed polyphenols such as pyrocatechol, gallic acid, resorcinol, catechin, chlorogenic acid, caffeic acid, syringic acid, p-coumaric acid, ferulic acid, vanillic acid, coumarin, sinapinic acid, trans-cinnamic acid, rutin, quercetin and kaempferol. The brine shrimp lethality assay ranked 78.60% extracts as cytotoxic (LC ≤ 250 μg/ml) whereas significant THP-1 inhibition was shown by methanol-acetone extract (IC 4.70 ± 0.43 μg/ml). The antiproliferative activity against Hep-G2 hepatoma cancer cell line was demonstrated by n-hexane, ethylacetate and methanol-distilled water (IC 8.65-8.95 μg/ml) extracts. Methanol extract displayed prominent protein kinase inhibitory activity (MIC 12.5 μg/disc) while n-hexane extract revealed remarkable antileishmanial activity (IC 4.69 ± 0.01 μg/ml). The antidiabetic potential was confirmed by n-hexane extract (44.70 ± 0.30% α-amylase inhibition at 200 μg/ml concentration) while a moderate antibacterial and antifungal activities were unveiled.
The variation in biological spectrum resulted due to use of multiple solvent systems for extraction. We also deduce that the valuable information gathered can be utilized for discovery of anticancer, antileishmanial, antioxidant and antidiabetic bioactive lead candidates.
由于植物源药物的广泛重要性，植物疗法的概念得以复兴。因此，对筋骨草的改善特性进行了研究。
采用比色法测定总酚含量、黄酮含量、抗氧化能力、还原能力和自由基清除活性。通过反相高效液相色谱（RP-HPLC）分析对特定多酚进行定量。使用卤虫致死试验测试初步细胞毒性，同时分别通过MTT和SRB方案测定对THP-1和Hep-G2细胞系的抗增殖活性。通过MTT比色法评估抗利什曼原虫潜力。为研究抗糖尿病前景，采用α-淀粉酶抑制试验，而采用纸片扩散法检测可能的蛋白激酶抑制、抗菌和抗真菌活性。
在15种不同提取物中，甲醇提取物表现出最高的总酚含量（10.75±0.70μg没食子酸当量/mg干重）、总还原能力（23.90±0.70μg抗坏血酸当量/mg干重）和总抗氧化能力（11.30±0.80μg抗坏血酸当量/mg干重），且提取物回收率最高（17.50±0.80% w/w）。氯仿-甲醇提取物表现出最高的黄酮含量（4.10±0.40μg槲皮素当量/mg干重），乙醇提取物表现出最强的自由基清除活性（IC50为14.40±0.20μg/ml）。基于RP-HPLC的定量分析证实了多种多酚的存在，如邻苯二酚、没食子酸、间苯二酚、儿茶素、绿原酸、咖啡酸、丁香酸、对香豆酸、阿魏酸、香草酸、香豆素、芥子酸、反式肉桂酸、芦丁、槲皮素和山奈酚。卤虫致死试验将78.60%的提取物列为具有细胞毒性（LC50≤250μg/ml），而甲醇-丙酮提取物对THP-1表现出显著抑制作用（IC50为4.70±0.43μg/ml）。正己烷、乙酸乙酯和甲醇-蒸馏水提取物（IC50为8.65 - 8.95μg/ml）对Hep-G2肝癌细胞系表现出抗增殖活性。甲醇提取物表现出显著的蛋白激酶抑制活性（MIC为12.5μg/纸片），而正己烷提取物显示出显著的抗利什曼原虫活性（IC50为4.69±0.01μg/ml）。正己烷提取物证实具有抗糖尿病潜力（在200μg/ml浓度下α-淀粉酶抑制率为44.70±0.30%），同时还显示出中等程度的抗菌和抗真菌活性。
由于使用多种溶剂系统进行提取，导致生物活性谱存在差异。我们还推断，所收集的有价值信息可用于发现抗癌、抗利什曼原虫、抗氧化和抗糖尿病生物活性先导化合物。
