金黄色葡萄球菌SsbA蛋白(SaSsbA)是一种单链DNA结合蛋白(SSB),是DNA复制和细胞存活所必需的,因此,它是潜在的抗病原体化疗的有吸引力的靶标。在这项研究中,我们制备了从100%丙酮中提取的紫癜的茎提取物,以研究其对SaSsbA的抑制作用。此外,这种提取物对存活的细胞毒性作用,凋亡,扩散,还检查了B16F10黑素瘤细胞的迁移。最初,杨梅素,槲皮素,山奈酚,二氢槲皮素,二氢山奈酚,芦丁,儿茶素,β-amyrin,冬凌草甲素,硫黄素T,报春碱,和硫黄素S被用作针对SaSsbA的可能抑制剂。在这些化合物中,二氢山奈酚和冬凌草甲素能够抑制SaSsbA的ssDNA结合活性,IC50值分别为750±62和2607±242μM。鉴于二氢山奈酚和冬凌草甲素的抑制作用较差,我们筛选了紫癜的提取物,米兰达,和Pliniacauliflora用于SaSsbA抑制剂。紫藤茎提取物具有较高的抗SaSsbA活性,IC50值为4.0±0.3μg/mL。使用气相色谱-质谱法鉴定了紫癜茎提取物中最丰富的化合物。该提取物中最丰富的前五名是driman-8,11-二醇,脱氧丝胶内酯,stigmast-5-en-3-ol,apocynin,和α-amyrin.使用MOE-Dock工具,这些化合物的结合模式,以及二氢山奈酚和冬凌草甲素,对SaSsbA进行了阐明,并计算了它们的结合能。根据S分数,这些化合物的结合能力依次为:脱氧丝氨酸内酯>二氢山奈酚>apocynin>driman-8,11-diol>stigmast-5-en-3-ol>冬凌草甲素>α-amyrin。将B16F10细胞与浓度为100μg/mL的紫癜茎提取物一起孵育,导致死亡率为76%,迁移减少了95%,抑制99%的增殖和集落形成,诱导细胞凋亡,这在96%的B16F10细胞中观察到。总的来说,本研究中的集体数据表明紫菜茎提取物在进一步医学应用中的药理潜力。
The Staphylococcus aureus SsbA protein (SaSsbA) is a single-stranded DNA-binding protein (SSB) that is categorically required for DNA replication and cell survival, and it is thus an attractive target for potential antipathogen chemotherapy. In this study, we prepared the stem extract of Sarracenia purpurea obtained from 100% acetone to investigate its inhibitory effect against SaSsbA. In addition, the cytotoxic effects of this extract on the survival, apoptosis, proliferation, and migration of B16F10 melanoma cells were also examined. Initially, myricetin, quercetin, kaempferol, dihydroquercetin,
dihydrokaempferol, rutin, catechin, β-amyrin, oridonin, thioflavin T, primuline, and thioflavin S were used as possible inhibitors against SaSsbA. Of these compounds,
dihydrokaempferol and oridonin were capable of inhibiting the ssDNA-binding activity of SaSsbA with respective IC50 values of 750 ± 62 and 2607 ± 242 μM. Given the poor inhibition abilities of
dihydrokaempferol and oridonin, we screened the extracts of S. purpurea, Nepenthes miranda, and Plinia cauliflora for SaSsbA inhibitors. The stem extract of S. purpurea exhibited high anti-SaSsbA activity, with an IC50 value of 4.0 ± 0.3 μg/mL. The most abundant compounds in the stem extract of S. purpurea were identified using gas chromatography−mass spectrometry. The top five most abundant contents in this extract were driman-8,11-diol, deoxysericealactone, stigmast-5-en-3-ol, apocynin, and α-amyrin. Using the MOE-Dock tool, the binding modes of these compounds, as well as
dihydrokaempferol and oridonin, to SaSsbA were elucidated, and their binding energies were also calculated. Based on the S scores, the binding capacity of these compounds was in the following order: deoxysericealactone >
dihydrokaempferol > apocynin > driman-8,11-diol > stigmast-5-en-3-ol > oridonin > α-amyrin. Incubation of B16F10 cells with the stem extract of S. purpurea at a concentration of 100 μg/mL caused deaths at the rate of 76%, reduced migration by 95%, suppressed proliferation and colony formation by 99%, and induced apoptosis, which was observed in 96% of the B16F10 cells. Overall, the collective data in this study indicate the pharmacological potential of the stem extract of S. purpurea for further medical applications.