Yun Keli, Bai Jian-Hai, Wang ZhenYu
Department of Food science and Engineering, School of Chemistry and Chemical Engineering, Harbin Institute of Technology, Harbin, P.R. China.
Department of Ophthalmology, North China University of Science and Technology Affiliated Hospital, Tangshan 064300, P.R. China.
Pharmacogn Mag. 2017 Oct-Dec;13(52):712-718. doi: 10.4103/pm.pm_409_16. Epub 2017 Nov 13.
The aim of this study was to evaluate radioprotective effect of extracts of bark and its fractions on rat splenocytes by using bioassay-guided isolation in order to obtain the best active fraction.
bark was ground and extracted with water, 40% acetone, 95% ethanol. Bio-guided assay was selected as an evaluation method to further fractionate radioprotective component from bark extract. Total phenolic and flavonoid contents in fractions were also measured. Rat splenocytes were prepared by using mechanical trituration method. DNA damage was assessed as comet parameters (tail DNA%, tail length, tail moment, olive tail moment). The levels of malondialdehyde (MDA), and activity of superoxide dismutase (SOD), catalase (CAT) in cultured rat splenocytes were also measured.
The radioprotective effects decreased from rutin >95% ethanol extracts of bark (95EEP) >40AEP > WEP. The stimulating effects decreased from rutin > n-butanol extract (NBE) > EAE. The results demonstrate that there exists toxic ingredients (PEE and dichloromethane extract), proliferative-promoting, radioprotective component (EAE and NBE) in 95EEP. fraction eluted from n-butanol fractions of 95EEP with 50% methanol solution (NBEPKB-50ME), a fraction of NBE result from bio-guided isolation, demonstrates good radioprotective efficacy on rat splenocytes. NBEPKB-50ME pretreated rat splenocytes demonstrated progressively reduced levels of MDA when compared with γ-ray exposed cells. Different dose of NBEPKB-50ME pretreatment with 8 Gy-irration showed an increase in enzymatic antioxidant.
Proliferative-promoting efficacy, radioprotective effect of different solvents extracts of the bark of were investigated in this work. NBEPKB-50ME was the best elution in NBE, especially in restoring SOD, CAT activities, content of GSH, decreasing DNA damage.
The radioprotective effects decreased from rutin > 95EEP > 40AEP > WEP. The extract of Petroleum ether, dichloromethane extract (DME) of 95% ethanol extract of (PEE, DME) show toxic effect on rat splenocytes. The extract of Ethyl acetate, n-butanol extract of 95% ethanol extract of (EAE, NBE) show proliferative-promoting, radioprotective effect on rat splenocytesSingle-cell gel electrophoresis was used to evaluate the spleen cell DNA damage parameters affected by gamma-radiation and addition of best component NBEPKB-50Me from extract of barkNBEPKB-50ME pretreatment with 8 Gy-irradiation showed an increase in enzymatic antioxidant capacity. NBEPKB-50ME pretreated (80, 160, 320, 480 mg/ml) rat splenocytes demonstrated progressively reduced levels of MDA when compared with g-ray exposed cells. MDA: Malondialdehyde; SOD: Superoxide dismutase; CAT: Catalase; PEE: Petroleum ether Extract; DME: Dichloromethane extract; EAE: Ethyl acetate extract; NBE: n-butanol extract; WAP: Water extracts of bark; 40AEP: 40% acetone extracts of bark; 95EEP: 95% ethanol extracts of bark; TPC: Total phenolic content; TFC: Total flavonoid content; NBEPKB-50ME: Fraction eluted from n-Butanol fractions of 95EEP with 50% methanol solution.
本研究旨在通过生物测定导向分离法评估[植物名称]树皮提取物及其各馏分对大鼠脾细胞的辐射防护作用，以获得最佳活性馏分。
将[植物名称]树皮研磨后分别用水、40%丙酮、95%乙醇进行提取。选择生物导向测定法作为评估方法，从[植物名称]树皮提取物中进一步分离辐射防护成分。同时测定各馏分中的总酚和黄酮含量。采用机械研磨法制备大鼠脾细胞。通过彗星参数（尾DNA%、尾长、尾矩、橄榄尾矩）评估DNA损伤情况。还测定了培养的大鼠脾细胞中丙二醛（MDA）水平、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）的活性。
辐射防护效果从芦丁> [植物名称]树皮95%乙醇提取物（95EEP）> 40%丙酮提取物（40AEP）>水提取物（WEP）依次降低。刺激作用从芦丁>正丁醇提取物（NBE）>乙酸乙酯提取物（EAE）依次降低。结果表明，95EEP中存在有毒成分（石油醚提取物和二氯甲烷提取物）、增殖促进和辐射防护成分（EAE和NBE）。用50%甲醇溶液从95EEP的正丁醇馏分中洗脱得到的馏分（NBEPKB - 50ME），这是生物导向分离得到的NBE的一个馏分，对大鼠脾细胞显示出良好的辐射防护效果。与γ射线照射的细胞相比，NBEPKB - 50ME预处理的大鼠脾细胞中MDA水平逐渐降低。不同剂量的NBEPKB - 50ME预处理8 Gy照射的细胞显示酶抗氧化剂增加。
本研究考察了[植物名称]树皮不同溶剂提取物的增殖促进效果和辐射防护作用。NBEPKB - 50ME是NBE中最佳洗脱馏分，尤其在恢复SOD、CAT活性、谷胱甘肽含量、减少DNA损伤方面表现突出。
辐射防护效果从芦丁> 95EEP > 40AEP > WEP依次降低。[植物名称]的石油醚提取物、95%乙醇提取物的二氯甲烷提取物（PEE、DME）对大鼠脾细胞显示出毒性作用。[植物名称]的乙酸乙酯提取物、95%乙醇提取物的正丁醇提取物（EAE、NBE）对大鼠脾细胞显示出增殖促进和辐射防护作用。采用单细胞凝胶电泳评估γ射线辐射及添加[植物名称]树皮提取物中最佳成分NBEPKB - 50Me对脾细胞DNA损伤参数的影响。8 Gy照射下NBEPKB - 50ME预处理显示酶抗氧化能力增加。与γ射线照射的细胞相比，NBEPKB - 50ME预处理（80、160、320、480 mg/ml）的大鼠脾细胞中MDA水平逐渐降低。MDA：丙二醛；SOD：超氧化物歧化酶；CAT：过氧化氢酶；PEE：石油醚提取物；DME：二氯甲烷提取物；EAE：乙酸乙酯提取物；NBE：正丁醇提取物；WAP：[植物名称]树皮水提取物；40AEP：[植物名称]树皮40%丙酮提取物；95EEP：[植物名称]树皮95%乙醇提取物；TPC：总酚含量；TFC：总黄酮含量；NBEPKB - 50ME：用50%甲醇溶液从95EEP的正丁醇馏分中洗脱得到的馏分
