Treatment of mice with stem bark extract of Aphanamixis polystachya reduces radiation-induced chromosome damage

Purpose: Normal tissue radiosensitivity is the major limiting factor in radiotherapy of cancer. The use of phytochemicals may reduce the adverse effects of radiation in normal tissue. The effect of ethyl acetate fraction of Aphanamixis polystachya (EAP) was investigated on the radiation-induced chromosome damage in the bone marrow cells of Swiss albino mice exposed to various doses of γ-radiation.

Materials and methods: The mice were divided into two groups, one group was exposed to 0, 1, 2, 3, 4 or 5 Gy of γ-radiation, while another group received 7.5 mg/kg body weight (BW) of EAP 1 h before exposure to 0, 1, 2, 3, 4 or 5 Gy of γ-radiation. Various asymmetrical chromosome aberrations were studied in the bone marrow cells of mice at 12, 24 or 48 h post-irradiation. To understand the mechanism of action of the free radical scavenging activity of 0, 5, 10, 20, 30, 40, 50, 60 or 70 μg/ml EAP, assays were carried out in vitro.

Results: Irradiation of mice to different doses of gamma radiation caused a dose dependent elevation in the frequency of aberrant cells and chromosome aberrations like chromatid breaks, chromosome breaks, dicentrics, acentric fragments and total aberrations at all the post-irradiation times studied. The maximum asymmetrical aberrations were scored at 24 h post-irradiation except chromatid breaks that were highest at 12 h post-irradiation. A maximum number of polyploid and severely damaged cells (SDC) were recorded at 24 h post-irradiation in the SPS + irradiation group. Treatment of mice with 7.5 mg/kg BW of EAP before exposure to 1 – 5 Gy of whole body gamma-radiation significantly reduced the frequencies of aberrant cells and chromosomal aberrations like acentric fragments, chromatid and chromosome breaks, centric rings, dicentrics and total aberrations at all post-irradiation scoring times (p < 0.01). The EAP showed a concentration dependent scavenging of hydroxyl, superoxide, 2,2′-diphenyl-1-picryl hydrazyl (DPPH) radicals and the 2,2-azino-bis-3-ethyl benzothiazoline-6-sulphonic acid (ABTS) cation radicals in vitro. EAP treatment also reduced lipid peroxidation in bone marrow cells in a concentration dependent manner.

Conclusion: Our study demonstrates that EAP protects mouse bone marrow cells against radiation-induced chromosomal aberrations and this reduction in radiation-induced chromosome damage may be due to free radical scavenging and reduction in lipid peroxidation. The radioprotection by EAP is best comparable to that of protection demonstrated by the grape fruit flavonone, naringin, in our earlier studies in mouse bone marrow cells.     

高效液相色谱法测定山核桃外蒲壳中4,8-二羟基-1-四氢萘酮的含量

目的采用高效液相色谱法(high performance liquid chromatography,HPLC)测定山核桃外蒲壳中4,8-二羟基-1-四氢萘酮的含量。方法采用HPLC,色谱柱为Shim-pack C18(250mm×4.6mm,5μm),以甲醇-水(40∶60)为流动相,流速为0.8ml/min,检测波长为333nm,柱温为25℃。结果 4,8-二羟基-1-四氢萘酮在1.13～20.34μg/ml范围内与峰面积呈良好线性关系,r=0.999 9,平均回收率为101.8%,相对标准偏差为1.21%(n=6)。结论所建立的HPLC准确可靠、结果稳定、重现性好,可用于山核桃外蒲壳4,8-二羟基-1-四氢萘酮的含量测定。     

高效液相色谱法测定山核桃外蒲壳中4,8-二羟基-1-四氢萘酮的含量

目的 采用高效液相色谱法(high performance liquid chromatography,HPLC)测定山核桃外蒲壳中4,8-二羟基-1-四氢萘酮的含量.方法 采用HPLC,色谱柱为Shim-pack C18 (250 mm×4.6 mm,5 μm),以甲醇-水(40∶60)为流动相,流速为0.8 ml/min,检测波长为333 nm,柱温为25 ℃.结果 4,8-二羟基-1-四氢萘酮在1.13～20.34 μg/ml范围内与峰面积呈良好线性关系,r=0.999 9,平均回收率为101.8%,相对标准偏差为1.21%(n=6).结论 所建立的HPLC准确可靠、结果稳定、重现性好,可用于山核桃外蒲壳4,8-二羟基-1-四氢萘酮的含量测定.     

山核桃叶球松素制备工艺研究

目的 优化山核桃叶球松素的制备工艺,为进一步开发利用山核桃叶提供理论依据。方法 采用单因素试验,山核桃叶通过乙醇回流提取,氢氧化钠变性,聚酰胺吸附并洗脱,石油醚萃取,乙醇结晶等步骤,以球松素作为检测指标,进行RP-HPLC测定。结果 山核桃叶球松素的制备工艺为山核桃叶粉末用10倍量的95%乙醇回流提取2次,每次30 min,以体积比400：5的比例往提取液中加入1 mol·L^-1氢氧化钠溶液,静置过夜取上清,以液质比20：1的比例与聚酰胺混合并旋蒸至干燥,先用15倍量聚酰胺体积的水洗去杂质,再用15倍量聚酰胺体积的20%乙醇溶液洗去杂质,最后用15倍量聚酰胺体积的60%乙醇溶液解吸附;收集60%乙醇溶液洗脱液,浓缩至干后,获得球松素含量在20%左右的粗产物,以液质比50：3的比例将此粗产物溶解于甲醇溶液中,加入等体积石油醚,连续萃取10次,收集石油醚产物,再将此产物用乙醇进行结晶,获得纯度>90%的球松素。结论 该方法稳定可行,适合工业生产;氢氧化钠变性、聚酰胺纯化及结晶是制备高纯度球松素的关键步骤,为球松素大规模制备奠定了基础,也为山核桃叶的开发利用提供了理论依据。     

Preliminary study of the anti-inflammatory activity of hexane extract and fractions from Bursera simaruba (Linneo) Sarg. (Burseraceae) leaves

Bursera simaruba (L.) Sarg. leaves hexane extracts display anti-inflammatory activity on the adjuvant-carrageenan-induced inflammation in rats. In order to isolate and identify the active compounds of the hexane extract, we performed a preliminary phytochemical study and a bioassay-directed fractionation using the carrageenan-induced paw oedema test in mice. From the nine fractions (A-I) obtained, of an initial chromatographic separation, fractions A and E (doses equivalents to 1.50 g dry plant/kg body weight) showed the strongest anti-inflammatory activity comparable to that of the reference drug phenylbutazone (80 mg/kg). The isolation and characterization of 3-methylene-7,11,15-trimethylhexadec-1-ene (neophytadiene) (1), ergost-5-en-3beta-ol (2), 24S-stigmast-5,22E-dien-3beta-ol (3), 24S-stigmast-5-en-3beta-ol (4) and alpha-amyrin (5), from these fractions is reported.     

HPLC同时测定山核桃外蒲壳中球松素和大黄酚含量

目的:建立同时测定山核桃外蒲壳中球松素和大黄酚含量的分析方法。方法:采用HPLC法,色谱柱为Shim-pack C18(250 mm×4.6 mm,5μm),以甲醇-0.1%磷酸水溶液(79∶21)为流动相,流速0.8 mL·min-1,检测波长286 nm,柱温25℃。结果:山核桃外蒲壳中球松素和大黄酚的线性范围分别为4.12～25.7μg·mL-1和1.01～13.4μg·mL-1,平均加样回收率(n=6)分别为98.8%和98.0%。结论:该方法准确可靠、结果稳定、重复性好,为评价山核桃外蒲壳的质量提供依据。     

Screening and Selection of Plants by Positive Pharmacologic Effect on Jejunum Muscular Contractility

Eight medicinal plant extracts (Bidens pilosa Linn, Eysenhardtia polystachya Sarg, Lepidium virginicum L., Litsea glaucescens H.B.K., Mirabilis jalapa Linn., Satureja macrostema Brieq., Sophora secundiflora Lag, and Tagetes micrantha Cav.) were tested for their effect on jejunum muscular contractility. All showed inhibitory activity and extracts of M. jalapa and S. macrostema exhibited highest activity with inhibitory concentrations (IC₅₀) of 18 and 73?µg/mL, respectively. Thus, we could corroborate some common traditional uses afforded to these plants. Flowers (petals, calices, and buds) of M. jalapa were the most active part of the plant. We obtained a semipurified sample of the flowers that was more or less five times more active than the crude extract. Furthermore, β-sitosterol, a compound reported as constitutive of M. jalapa, showed no effect on jejunum contractility.     

山核桃外蒲壳化学成分及抗肿瘤活性研究

目的:研究山核桃外蒲壳的化学成分和抗肿瘤活性。方法:采用硅胶柱色谱、Sephadex LH-20分离化学成分,根据理化性质和波谱数据确定化合物结构,并对得到的化合物进行细胞毒活性研究。结果:从山核桃外蒲壳中共分离鉴定了5个化合物,分别为:大黄酚(1)、大黄素甲醚(2)、β-谷甾醇(3)、球松素(乔松酮)(4)、4,8-二羟基-1-四氢萘酮(5)。结论:其中,化合物2、5为首次从山核桃属植物中分离得到;在MTT抗肿瘤实验中,化合物1、4、5均具有一定的抗肿瘤活性。     

山核桃叶中总黄酮含量测定

目的:建立山核桃叶中总黄酮的含量测定方法。方法:以槲皮苷为对照品,采用AlCl3显色紫外分光光度法,结合单因素试验和正交试验对该体系进行系统优化研究,测定山核桃叶中总黄酮的含量。结果:AlCl3显色最佳条件为:2.0mL体系中加入pH4.7的HAc-NaAc缓冲液0.5mL,10%AlCl3醇溶液0.4mL,蒸馏水0.1mL,常温反应20min,416 nm处测定吸光度值。槲皮苷标准品含量在2.6～14.3μg.mL-1范围内与吸光度呈良好的线性关系(r=0.998),测得充分提取的总黄酮平均含量为5.89%,RSD为1.14%(n=3),平均回收率为99.1%,RSD为1.27%(n=6)。结论:AlCl3显色紫外分光光度法操作简便、准确、灵敏、重现性好,可用于山核桃叶总黄酮含量的测定。     

Antidiarrheal and Antispasmodic Activities of Buddleja polystachya are Mediated Through Dual Inhibition of Ca++ Influx and Phosphodiesterase Enzyme

This study describes the antidiarrheal and antispasmodic activities of the hydro‐alcoholic extract of Buddleja polystachya (Bp.Cr) with possible mode of action explored along with activity‐directed fractionation. Bp.Cr and its aqueous (Bp.Aq) and organic fractions, petroleum ether (Bp.Pet), dichloromethane (Bp.DCM), ethylacetate (Bp.EtAc) and butanol (Bp.But), were tested using the in‐vivo and in‐vitro assays. The crude extract (100–300 mg/kg) showed 20 and 60% protection of castor oil‐induced diarrhea in mice. In isolated rabbit jejunum, Bp.Cr like papaverine inhibited spontaneous and high K⁺ (80 mM)‐induced contractions equi‐potently. In guinea‐pig ileum, Bp.Cr showed a moderate spasmogenic effect. The activity‐directed fractionation revealed that the spasmolytic activity was concentrated in the organic fractions and spasmogenic component in the aqueous fraction. Amongst the organic fractions, BP.DCM and Bp.Pet inhibited spontaneous and high K⁺‐induced contractions equi‐potently, while Bp.But, like verapamil was more potent against high K⁺. The crude extract and its organic fractions caused rightward shift in the Ca⁺⁺‐concentration response curves (CRCs), similar to verapamil, and all except Bp.But potentiated the isoprenaline‐inhibitory CRCs to the left, similar to papaverine. The results of this study indicate that the crude extract of B. polystachya possesses antidiarrheal and antispasmodic activities, mediated possibly through dual inhibition of Ca⁺⁺ influx and phospodiesterase enzyme. Copyright © 2015 John Wiley & Sons, Ltd.