苦参碱类生物碱的镇痛作用研究进展

苦参碱类生物碱（包括有苦参碱、氧化苦参碱、氧化槐定碱、槐果碱、氧化槐果碱等）具有镇痛作用,镇痛特征是以中枢性为主、无成瘾性和耐药性,综述其镇痛作用及作用机制研究进展。苦参碱类生物碱的中枢性镇痛作用机制可能是：激活大麻素受体-2和上调电压门控钙离子N-型通道表达,促进抑制性神经递质（γ-氨基丁酸、甘氨酸）合成和释放,下调GAT-1表达,使突触间隙γ-氨基丁酸浓度升高以及上调γ-氨基丁酸-A受体表达,从而增强γ-氨基丁酸能神经功能。γ-氨基丁酸能神经功能的增强可抑制兴奋性神经递质谷氨酸过度表达并下调NMDA受体和蛋白激酶Cγ表达,降低NMDA受体对谷氨酸的兴奋性,从而下调电压门控钙离子L-型通道的表达,抑制钙离子内流,阻滞CaMKⅡ/CREB通路,减轻炎症反应和产生镇痛作用。     

外翻肠囊法研究氧化苦参碱磷脂复合物的肠吸收特性

目的 通过对比研究氧化苦参碱及其磷脂复合物大鼠离体肠吸收情况,考察磷脂复合物对氧化苦参碱肠吸收的影响。方法 采用大鼠离体外翻肠囊模型研究氧化苦参碱磷脂复合物的肠吸收特性。HPLC测定不同剂量的氧化苦参碱及氧化苦参碱磷脂复合物在大鼠十二指肠、空肠、回肠和结肠内的累积吸收量。结果 氧化苦参碱磷脂复合物在不同肠段的吸收较氧化苦参碱均有提高,低剂量组氧化苦参碱磷脂复合物在空肠、结肠处的吸收较氧化苦参碱有显著提高;中剂量组氧化苦参碱磷脂复合物在十二指肠、空肠、结肠处的累积吸收量较氧化苦参碱有显著提高;高剂量组氧化苦参碱磷脂复合物在空肠的吸收较氧化苦参碱有显著提高。结论 与氧化苦参碱相比,磷脂复合物对氧化苦参碱在大鼠肠道的吸收有显著的促进作用。     

A Study on Isolation of Chemical Constituents from Sophora Flavescens Ait. and Their Anti-Glioma Effects

Background

Sophora flavescens Ait. is a traditional Chinese medicine with a long history in China. It is mainly used in the treatment of heat dysentery and similar ailments in the clinical. The objective of this paper was to isolate, purify and identify alkaloids from Sophora flavescens Ait. and to explore their inhibitory effects on C6 glioma cells.

Materials and Methods

Column chromatography, extraction and NMR spectroscopy were used to structurally identify the isolated compounds. MTT assay and flow cytometry were used to detect the inhibitory effect of matrine on C6 cells.

Results

Three compounds were isolated from Sophora flavescens Ait., namely matrine, oxymatrine and lupeol. Different concentrations of matrine solution all had inhibitory effects on growth of C6 cell lines, which showed apparent dose-effect relationship. Compared with the control group, proportion of G0/G1 phase cells increased in each matrine concentration group to a maximum of 79.8%; proportion of S phase cells reduced, and proportion of G2/M phase cells declined slightly to a minimum of 6.3%, suggesting that after the action of matrine proliferation of C6 cells was significantly inhibited and the cells were arrested in the G1 phase.

Conclusion

We concluded that Sophora flavescens Ait. has an inhibitory effect on C6 cell proliferation.     

Extraction and Isolation of Alkaloids of Sophora Alopecuroides and Their Anti-Tumor Effects in H22 Tumor-Bearing Mice

Background

Alkaloids of Sophora alopecuroides have good biological activity, and are widely used in clinical settings, which not only have pharmacological activities of anti-cancer, cancer suppression, as well as the inhibition, and killing of various microorganisms; but also possess extensive pharmacological effects on immune system, nervous system and cardiovascular system. The objective of this paper was to extract and isolate total alkaloids of Sophora alopecuroides (TASA), and to study their anti-tumor effects in H22 tumor-bearing mice.

Materials and Methods

TASA were extracted and isolated using thin-layer chromatography, and column chromatography; and the isolated compounds were analyzed using nuclear magnetic resonance. The inhibitory effects of TASA on tumor in H22-bearing mice were determined by MTT assay.

Results

Three compounds were isolated from Sophora alopecuroides L., which were matrine, oxymatrine and sophoridine, respectively. Meanwhile, mouse H22 sarcoma model was established and different doses of TASA apparently inhibited solid H22-tumor in mice; it inhibited the thymus, and spleen to some extent; the degree of inhibition was more obvious for the spleen.

Conclusion

TASA has an anti-tumor effect in H22 tumor-bearing mice.     

氧化苦参碱对小鼠接触性皮炎的抑制作用

目的:探讨氧化苦参碱(OMT)对小鼠变态反应性接触性皮炎(ACD)的抑制作用.方法:56只BALB/c小鼠随机分成7组,每组8只.除B组(该组小鼠不致敏,也不诱发ACD)外,其他6组均建立ACD模型.A组每d腹腔注射20 ml/kg的PBS,B、C、D、E、F、G组分别每d腹腔注射OMT 50 mg/kg、12.5 mg/kg、25 mg/kg、50 mg/kg和100 mg/kg及氢化可的松25 mg/kg,均给药28 d.于给药第8 d、14 d、29 d测各组小鼠左、右耳厚度,计算2耳差值.实验第1 d、7 d、14 d、21 d、28 d小鼠尾静脉采血三色免疫荧光标记,以流式细胞仪检测各组CD4 CD25 T细胞数.第29 d处死全部小鼠,测量胸腺及脾脏质量.结果:OMT具有强烈抑制DNFB诱发的小鼠耳部ACD作用,且OMT可增加各剂量组小鼠外周血中CD4 CD25 T细胞数,呈剂量依赖性.小剂量的OMT可以使小鼠胸腺和脾脏质量增加,而大剂量的OMT对小鼠脾脏质量的影响不明显,但能降低小鼠胸腺的质量.结论:OMT显著升高小鼠外周血中CD4 CD25 T细胞数,可能是OMT抑制ACD的免疫学机制之一.     

苦参碱和氧化苦参碱对小鼠免疫功能的实验研究

[目的]探讨苦参碱和氧化苦参碱对免疫功能的作用。[方法]采用对T淋巴细胞酯酶染色率和溶血素抗体生成的方法。[结果]苦参碱和氧化苦参碱均可明显减少T淋巴细胞酯酶染色率;但可明显促进溶血素抗体形成。[结论]苦参碱和氧化苦参碱可明显降低T淋巴细胞功能,对体液免疫功能呈增强作用。     

磨积颗粒质量标准的研究

目的建立磨积颗粒的质量标准。方法建立黄芪、苦参的薄层色谱法鉴别方法和苦参碱及氧化苦参碱的高效液相色谱测定方法。结果定性方法能检出黄芪和苦参,且阴性对照无干扰;苦参碱进样量在0.312~0.936μg范围内、氧化苦参碱进样量在0.471~1.412μg范围内呈良好的线性关系,苦参碱平均加样回收率为97.53%,RSD=1.16%,氧化苦参碱平均加样回收率为98.02%,RSD=0.94%。结论该法操作简便、结果准确,重现性好,适用于磨积颗粒的质量控制。     

阳离子树脂分离氧化苦参碱工艺条件研究

目的:优化阳离子树脂分离氧化苦参碱的工艺条件,提高氧化苦参碱得率。方法:通过单因素试验考察了洗脱溶剂种类、洗脱溶液浓度、树脂碱化用氨水pH值等条件对氧化苦参碱得率的影响,并通过正交试验对工艺条件进行优化。结果:最适洗脱溶剂为乙醇溶液,最佳工艺条件为吸附后树脂用等量的pH值为12.5的氨水碱化,用12倍量70%乙醇洗脱。结论:采用最佳工艺条件氧化苦参碱得率由3.83mg/g提高到8.31mg/g,较优化前提高了2.17倍。     

高效液相色谱法测定当归苦参丸中苦参碱和氧化苦参碱

目的建立测定当归苦参丸中苦参碱和氧化苦参碱含量的高效液相色谱法。方法采用Phenemonex C18（250mm×4．6mm，5μm）色谱柱，流动相：乙腈-0．1％三氟醋酸溶液（5：95），流速：1．0mL·min^-1，检测波长：208nm，柱温：35℃。结果苦参碱和氧化苦参碱线性范围分别为0．0512～0．3584μg（r=0．9990）和0．1924～1．3468μg（r=0．9999）；平均回收率分别为98．73%,100．14％（n=6）。结论该方法操作简单，重现性好，检测灵敏度、准确度高，且样品分离效果好。适用于当归苦参丸中苦参碱和氧化苦参碱的含量测定。     

HPLC法测定苦参总碱中苦参碱与氧化苦参碱的含量

目的:采用高效液相色谱法测定苦参总碱中苦参碱和氧化苦参碱的含量。方法:采用 Elite Hypersil NH_2柱(5μm,250mm×4.6 mm),乙腈-无水乙醇-3%磷酸溶液(81:10:9)为流动相,流速:1.0 mL·min~(-1),检测波长220 nm,柱温:25℃,进样量:10 μL。结果:苦参碱、氧化苦参碱的线性范围分别为5.32～53.2μg·mL~(-1)(r=0.9997),65.7～525.6μg·mL~(-1)(r=0.9995)。苦参碱平均加样回收率为99.5%,RSD=1.0%(n=6);氧化苦参碱的平均加样回收率为99.7%,RSD=0.6%(n=6)。结论:本方法操作简便、快速、可靠,可用于控制苦参总碱质量。