从喜马拉雅雪松松针中提取和纯化总黄酮有助于体外抗肿瘤。

Extraction and purification of total flavonoids from pine needles of Cedrus deodara contribute to anti-tumor in vitro.

作者信息

Shi Xiaofeng, Liu Dongyan, Zhang Junmin, Hu Pengbin, Shen Wei, Fan Bin, Ma Quhuan, Wang Xindi

机构信息

Gansu Provincial Academy of Medical Sciences, Lanzhou, 730050, China.

Key Laboratory of TCM Pharmacology and Toxicology of Gansu Province, Lanzhou, 730030, China.

出版信息

BMC Complement Altern Med. 2016 Jul 26;16:245. doi: 10.1186/s12906-016-1249-z.

DOI:10.1186/s12906-016-1249-z

PMID:27461104

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4962484/

Abstract

BACKGROUND

Cedrus deodara is one of the traditional Chinese medicinal herbs that exhibits a line of biological activities. The current study extracted the total flavonoids from the pine needles of Cedrus deodara (TFPNCD), and investigated its anti-cancer effects in tumor cell lines.

METHODS

The total flavonoids was extracted from pine needles of Cedrus deodara by ethanol hot refluxing and purified by HPD722 macroporous resin. The contents of total flavonoids and the active ingredients of TFPNCD were analyzed through UV and HPLC. MTT assay was used to investigate its inhibitory effect on tumor cell lines. The flow cytometry was employed to determine the apoptosis and cell cycle distribution after treated TFPNCD on HepG2 cells.

RESULTS

The TFPNCD, in which the contents of total flavonoid reached up to 54.28 %, and the major ingredients of myricetin, quercetin, kaempferol and isorhamnetin in TFPNCD were 1.89, 2.01, 2.94 and 1.22 mg/g, respectively. The MTT assays demonstrated that TFPNCD inhibited the growth of HepG2 cells in a dose-dependent manner, with the IC50 values of 114.12 μg/mL. By comparison, TFPNCD inhibited the proliferation to a less extent in human cervical carcinoma HeLa, gastric cancer MKN28 cells, glioma SHG-44 cells and lung carcinoma A549 than HepG2 cells. We found that even at the lower doses, the total flavonoids effectively inhibited the proliferation of HepG2 cells. Comparison of IC50 values implicated that HepG2 cells might be more sensitive to the treatment with total flavonoids. TFPNCD was able to increase the population of HepG2 cells in G0 /G1 phase. Meanwhile, TFPNCD treatment increased the percentage of apoptotic HepG2 cells.

CONCLUSION

These data suggested that TFPNCD might have therapeutic potential in cancer through the regulation of cell cycle and apoptosis.

摘要

背景

雪松是具有一系列生物活性的传统中草药之一。本研究从雪松松针中提取了总黄酮（TFPNCD），并研究了其在肿瘤细胞系中的抗癌作用。

方法

通过乙醇热回流法从雪松松针中提取总黄酮，并用HPD722大孔树脂进行纯化。通过紫外分光光度法和高效液相色谱法分析TFPNCD中总黄酮的含量和活性成分。采用MTT法研究其对肿瘤细胞系的抑制作用。利用流式细胞术检测TFPNCD处理HepG2细胞后的凋亡情况和细胞周期分布。

结果

TFPNCD中总黄酮含量高达54.28%，其中杨梅素、槲皮素、山奈酚和异鼠李素的主要成分含量分别为1.89、2.01、2.94和1.22 mg/g。MTT分析表明，TFPNCD以剂量依赖性方式抑制HepG2细胞的生长，IC50值为114.12 μg/mL。相比之下，TFPNCD对人宫颈癌HeLa细胞、胃癌MKN28细胞、胶质瘤SHG-44细胞和肺癌A549细胞增殖的抑制程度低于对HepG2细胞的抑制程度。我们发现，即使在较低剂量下，总黄酮也能有效抑制HepG2细胞的增殖。IC50值的比较表明，HepG2细胞可能对总黄酮治疗更敏感。TFPNCD能够增加HepG2细胞在G0/G1期的比例。同时，TFPNCD处理增加了HepG2细胞的凋亡百分比。