Liu Xi, Zhu Liancai, Tan Jun, Zhou Xuemei, Xiao Ling, Yang Xian, Wang Bochu
Key Laboratory of Biorheological Science and Technology (Chongqing University), Ministry of Education, College of Bioengineering, Chongqing University, No, 174, Shapingba Main Street, Chongqing 400030, China.
BMC Complement Altern Med. 2014 Jan 10;14:12. doi: 10.1186/1472-6882-14-12.
In Chinese traditional medicine, Agrimonia pilosa Ledeb (APL) exhibits great effect on treatment of type 2 diabetes mellitus (T2DM), however its mechanism is still unknown. Considering that T2DM are correlated with postprandial hyperglycemia and oxidative stress, we investigated the α-glucosidase inhibitory activity and the antioxidant activity of flavonoid compound (FC) and triterpenoid compound (TC) from APL.
Entire plants of APL were extracted using 95% ethanol and 50% ethanol successively. The resulting extracts were partitioned and isolated by applying liquid chromatography using silica gel column and Sephadex LH 20 column to give FC and TC. The content of total flavonoids in FC and the content of total triterpenoids in TC were determined by using UV spectrophotometry. HPLC analysis was used to identify and quantify the monomeric compound in FC and TC. The α-glucosidase inhibitory activities were determined using the chromogenic method with p-nitrophenyl-α-D-glucopyranoside as substrate. Antioxidant activities were assessed through three kinds of radical scavenging assays (DPPH radical, ABTS radical and hydroxyl radical) & β-carotene-linoleic acid assay.
The results indicate FC is abundant of quercitrin, and hyperoside, and TC is abundant of 1β, 2β, 3β, 19α-tetrahydroxy-12-en-28-oic acid (265.2 mg/g) and corosolic acid (100.9 mg/g). The FC & the TC have strong α-glucosidase inhibitory activities with IC50 of 8.72 μg/mL and 3.67 μg/mL, respectively. We find that FC show competitive inhibition against α-glucosidase, while the TC exhibits noncompetitive inhibition. Furthermore, The FC exhibits significant radical scavenging activity with the EC50 values of 7.73 μg/mL, 3.64 μg/mL and 5.90 μg/mL on DPPH radical, hydroxyl radical and ABTS radical, respectively. The FC also shows moderate anti-lipid peroxidation activity with the IC50 values of 41.77 μg/mL on inhibiting β-carotene bleaching.
These results imply that the FC and the TC could be responsible for the good clinical effects of APL on T2MD through targeting oxidative stress and postprandial hyperglycaemia. So APL may be good sources of natural antioxidants and α-glucosidase inhibitors exhibiting remarkable potential value for the therapy of T2DM.
在中医中,仙鹤草对2型糖尿病(T2DM)的治疗具有显著疗效,但其作用机制尚不清楚。鉴于T2DM与餐后高血糖和氧化应激相关,我们研究了仙鹤草中黄酮类化合物(FC)和三萜类化合物(TC)的α-葡萄糖苷酶抑制活性和抗氧化活性。
依次用95%乙醇和50%乙醇提取仙鹤草全草。所得提取物通过硅胶柱和葡聚糖LH 20柱进行液相色谱分离,得到FC和TC。采用紫外分光光度法测定FC中总黄酮含量和TC中总三萜含量。用HPLC分析鉴定和定量FC和TC中的单体化合物。以对硝基苯基-α-D-吡喃葡萄糖苷为底物,采用显色法测定α-葡萄糖苷酶抑制活性。通过三种自由基清除试验(DPPH自由基、ABTS自由基和羟基自由基)及β-胡萝卜素-亚油酸试验评估抗氧化活性。
结果表明,FC富含槲皮苷和金丝桃苷,TC富含1β,2β,3β,19α-四羟基-12-烯-28-酸(265.2mg/g)和科罗索酸(100.9mg/g)。FC和TC具有较强的α-葡萄糖苷酶抑制活性,IC50分别为8.72μg/mL和3.67μg/mL。我们发现FC对α-葡萄糖苷酶表现出竞争性抑制,而TC表现出非竞争性抑制。此外,FC对DPPH自由基、羟基自由基和ABTS自由基的清除活性显著,EC50值分别为7.73μg/mL、3.64μg/mL和5.90μg/mL。FC在抑制β-胡萝卜素漂白方面也表现出中等程度的抗脂质过氧化活性,IC50值为41.77μg/mL。
这些结果表明,FC和TC可能通过靶向氧化应激和餐后高血糖,对仙鹤草治疗T2MD的良好临床效果起作用。因此,仙鹤草可能是天然抗氧化剂和α-葡萄糖苷酶抑制剂的良好来源,在T2DM治疗中具有显著的潜在价值。