石菖蒲水提物对PC12细胞增殖及细胞突起的作用

目的研究石菖蒲水提物对PC12细胞增殖及细胞突起的作用,观察药物是否具有神经营养功能或促进细胞分化的功能。方法采用细胞计数试剂盒检测不同浓度的石菖蒲水提物对细胞活力的影响;采用Image J软件统计PC12细胞的梭型细胞率、细胞分化率、细胞突起长度及阳性细胞分化率,以评价石菖蒲水提物对细胞分化的影响。结果石菖蒲水提物在完全培养条件和低血清培养条件下对PC12细胞增殖有显著的促进作用,对细胞突起的长出、分支、延伸有明显的抑制作用。结论石菖蒲水提物具有一定的神经营养功能。     

Inhibitory effects of Acorus calamus extracts on mast cell-dependent anaphylactic reactions using mast cell and mouse model

Ethnopharmacological relevance

Acorus calamus Linn. (Araceae) is a traditional herbal plant used for centuries to treat various allergic symptoms including asthma and bronchitis.

Aim of the study

The present study was focused to provide a pharmacological basis for the traditional use of Acorus calamus in allergic symptoms using the mast cell-dependent anaphylactic reactions in in vitro and in vivo models.

Materials and methods

Cell viabilities were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Dinitrophenyl-human serum albumin (DNP-HSA) induced β-hexosaminidase and interleukin (IL)-4 productions in IgE-sensitized rat basophilic leukaemia (RBL-2H3) cells were measured by enzymatic assay and enzyme-linked immunosorbent assay (ELISA). Passive cutaneous anaphylaxis (PCA) reaction mouse model was implemented for in vivo studies.

Results

Hot water (HW), butylene glycol (BG), hexane (HE) and steam distilled (SD) extracts of Acorus calamus showed different cytoxicity levels evaluated in RBL-2H3 cells. Sub-toxic doses of HW extract suppressed the β-hexosaminidase secretion and IL-4 production significantly and dose dependently in DNP-HSA induced IgE-sensitized RBL-2H3 cells compared to other extracts of Acorus calamus. Further, in vivo studies also revealed that the HW extract significantly inhibited the PCA reaction in mouse compared to the normal control group.

Conclusion

HW extract of Acorus calamus most effectively inhibited degranulation and IL-4 secretion in DNP-HSA-stimulated RBL-2H3 cells and also reduced the mast cell-mediated PCA reaction in mouse, providing a therapeutic evidence for its traditional use in ameliorating allergic reactions.     

石菖蒲内生真菌灰黄青霉Z005次级代谢产物的研究

目的 研究药用植物石菖蒲内生真菌灰黄青霉Penicillium griseofulvum Z005的次级代谢产物。方法 利用包括硅胶、ODS、Sephadex LH-20、半制备HPLC等色谱方法,对目标菌株的大米固体发酵提取物进行系统的分离纯化,并结合理化常数与现代的谱学方法进行化合物的结构鉴定。结果 从石菖蒲内生真菌灰黄青霉Z005固体发酵产物中分离鉴定了1个新化合物和12个已知化合物。分别为灰黄青霉呋喃醇A(1)、chaetominine(2)、dehydrocyclopeptine(3)、1-methylcyclopenin(4)、4-(2-羟基丁炔氧基)苯甲酸（5）、penipratynolene(6)、hypofuran B(7)、lobechine(8)、4-(2-甲酰基-5-甲氧基甲基-1H-吡咯-1-基)丁酸（9）、2-呋喃甲醇-(5’→11)-1,3-环戊二烯-[5,4-c]-1H-邻二氮杂萘（10）、N-乙酰色胺（11）、1H-吲哚-3-基甲基氨基甲酸甲酯（12）和顺式阿魏酸甲酯（13）。结论 灰黄青霉Z005为首次从石菖蒲健康组织中分离获得,化合物1为新的苯并呋...     

石菖蒲加冰片对脑皮质神经细胞保护作用及其伍用配比的研究

目的 观察石菖蒲加冰片不同配比对新生大鼠脑皮质神经细胞的保护作用，从而筛选出较佳配比。方法 培养脑皮质神经细胞，造缺氧模型，观察给药组与对照组的bcl-2、bax、caspase-3、白细胞介素-6(IL-6)等指标及原位细胞凋亡情况。结果 药物组能降低促凋亡基因caspase-3、bax的表达；而提高抑制凋亡基因bcl-2的表达，减少脑皮质神经细胞凋亡；减少IL-6的生成。石菖蒲加冰片不同配比明显比单纯石菖蒲挥发油组和单纯冰片组的作用效果好，其中高、中、低配比组之间比较，高配比组作用效果最好。结论 石菖蒲加冰片能减少脑皮质神经细胞凋亡，对脑皮质神经细胞有保护作用，尤以高配比组(冰片：挥发油=2：1．46)作用更明显，为较佳配伍。     

固相微萃取技术/气相色谱/质谱分析水菖蒲挥发性化学成分

目的:分析水菖蒲挥发性化学成分。方法:利用固相微萃取/气相色谱/质谱(SPME/GC/MS)联用技术对水菖蒲挥发性成分进行研究。结果:共鉴定出69种化学成分,占挥发性总成分的81.75%。结论:水菖蒲主要挥发性化学成分是1,4-顺-1,7-顺-菖蒲烯酮(11.72%),2-β-异丙烯基-6-α-异丙烯基-3-β-甲基-3-α-乙烯基环己酮(6.04%),β-倍半水芹烯(5.71%),α-佛手柑油烯(4.58%),γ-紫穗槐烯(4.11%),γ-杜松烯(4.07%),顺-甲基-异丁香油酚(4.03%)等。     

优选细辛醚组合对β-淀粉样蛋白诱导PC12细胞损伤的保护作用

目的筛选石菖蒲有效成分α-细辛醚、β-细辛醚、丁香酚保护PC12细胞免受β-淀粉样蛋白毒性损伤作用的优化组合及其剂量。方法采用正交实验设计,以细胞存活率和凋亡率为指标,筛选3种成分联合作用的优化比例,并探讨此组合的优化剂量。结果优化组合为:α-细辛醚为0μM、β-细辛醚为10μM、丁香酚为3μM,较β-细辛醚、丁香酚单独发挥作用的所需浓度低(仅为各自单个成分剂量的1/5、1/20,但已能发挥显著作用)。结论石菖蒲有效成分的适当配比,能更有效地保护PC12细胞免受β淀粉样蛋白毒性损伤。     

石菖蒲挥发油香熏辅助治疗老年性痴呆大鼠的实验研究

采用D-乳糖和氯化铝致大白鼠老年性痴呆模型,分别用脑复康、脑复康加石菖蒲挥发油治疗观察,结果显示石菖蒲挥发油加脑复康能明显改善模型大鼠学习记忆能力,提高脑组织重量、提高脑组织超氧化物歧化酶含量、血清睾酮含量,降低模型大鼠血清乙酰胆硷脂酶水平、脑组织丙二醛含量.其疗效优于对照组和单纯脑复康组.提示石菖蒲外嗅香熏辅助治疗对于阻止脑萎缩,防治老年性痴呆具有重要的作用.     

羌菖合剂对右脑供血不足大鼠外周血红细胞的影响

目的 研究羌菖合剂对脑供血不足大鼠外周红细胞的作用,探讨其改善血液流变性的原理。方法 手术结扎大白鼠右侧颈总动脉造成脑供血不足;术后3、7天仪器分析红细胞及其流变性参数。结果 (1)术后3d,对照组大鼠外周血红细胞数(RBC)、血红蛋白(HGB)和平均红细胞体积(MCV)与假术组无明显差异,平均细胞血红蛋白(MCH)、血球比积(HCT)和RBC聚集指数呈增高趋势,平均细胞血红蛋白浓度(MCHC)明显增大(P<0.05);羌菖合剂10g/kg治疗3d可显著防止上述改变(P<0.05～P<0.001)。(2)术后7d,对照组外周血红细胞及其血红蛋白量均比假术组无明显差异,但RBC刚性指数和聚集指数则呈进一步升高趋势;羌菖合剂治疗组MCHC、RBC刚性指数和聚集指数均显著低于对照组(P<0.05)。结论 羌菖合剂治疗能防止脑供血不足大鼠外周血红细胞改变;降低MCHC,降低RBC刚性指数和聚集指数,从而改善RBC变形能力和流变性。     

Protective effect of acorus calamus against acrylamide induced neurotoxicity

Exposure of rats to acrylamide (ACR) caused hind limb paralysis in 58% of the animals on day 10 and decreased behavioural parameters, namely distance travelled, ambulatory time, stereotypic time and basal stereotypic movements compared with the control group. These rats also had a decrease in the reduced glutathione (GSH) content and glutathione-S-transferase (GST) activity in the corpus striatum and an increase in striatal dopamine receptors, as evident by an increase in the binding of 3H-spiperone to striatal membranes. Treatment with the ethanol:water (1:1) extract of the rhizomes of Acorus calamus (AC-002) increased the GSH content and GST activity in the corpus striatum while insignificant changes were observed in other parameters. Rats treated with ACR and AC-002 in combination had a lower incidence of paralysis (18%) compared with those treated with ACR alone on day 10 of the experiment. The rats also showed a partial recovery in other behavioural parameters. The levels of GSH content and GST activity increased in the corpus striatum, while the dopamine receptors decreased compared with the ACR treated rats. The results suggest that the neurobehavioural changes produced by ACR may be prevented following treatment with Acorus calamus rhizomes.     

石菖蒲对缺血再灌注脑损伤大鼠神经细胞凋亡的影响

目的 :观察石菖蒲对缺血再灌注脑损伤大鼠神经细胞凋亡影响的有效部位。方法 :提取石菖蒲 (去油水提液、含油水提液、挥发油、β -细辛醚 ) ,灌胃给药 7d ,1次 /d ,观察大鼠脑皮质和海马神经细胞凋亡的情况。结果 :β -细辛醚有明显抑制大鼠脑皮质神经细胞凋亡的作用 ,石菖蒲组分 (含油水提液、挥发油、β -细辛醚 )也有减少大鼠海马神经细胞凋亡的作用。β -细辛醚能抑制大鼠脑皮质和海马神经细胞Bax基因表达。石菖蒲挥发油和 β -细辛醚能增强大鼠脑皮质神经细胞Bcl -X基因的表达。结论 :石菖蒲挥发油尤其是 β -细辛醚有抑制大鼠神经细胞凋亡的作用