桔梗叶绿体比较基因组学分析及系统发育研究

目的 以桔梗Platycodon grandiflorus为材料,分析其叶绿体基因组特征,探究不同地区桔梗叶绿体基因组的差异及桔梗科其他物种的系统发育关系。方法 利用Illumina NovaSeq测序平台对桔梗叶绿体全基因组进行测序,完成其组装、注释和特征分析,采用生物信息学方法对不同地区桔梗进行比较基因组分析和系统发育分析。结果 桔梗叶绿体基因组全长172 770 bp,呈现典型的环状四分体结构,总GC含量为38.10%,注释到139个基因,其中蛋白质编码基因95个,核糖体RNA 8个和转运RNA 36个。经序列分析鉴定出139个SSR位点,大部分重复由A和T组成。该叶绿体基因组密码子偏好性A/U大于G/C。边界分析表明,不同地区桔梗的JLA边界区域存在差异。对比不同地区桔梗叶绿体基因组序列发现21个变异区间,包括ycf1、psbC、rps18和rpoB等编码区以及rpl32-trnL、trnS-psbZ和trnN-ycf1等非编码区。基于最大似然法（maximum likelihood method,ML）对桔梗及其他17种桔梗科植物进行系统发育分析,发现桔梗科物种形成一个单系群,各属物种聚为一束,支持率达100%。结论 桔梗科物种聚为一支与传统相符合,不同地区桔梗叶绿体基因组序列存在显著差异,为后期开展分子鉴定及群体遗传学研究提供提供科学依据。     

桔梗扦插繁殖试验

对桔梗进行扦插繁殖试验，结果看出：用茎基部作插条的成活率最高（５２．２３％），茎梢部作插条成活率很低（１．１１％）；用激素处理可大大提高插条的成活率，其中ＮＡＡ（萘乙酸）１００ｐｐｍ处理３ｈ最显著，比对照组提高２３．３３％；扦插基质湿度过大，会引起插条腐烂。     

一年生桔梗质量研究初探

为了促进中药材GAP的发展，本文采用中国药典法测定了不同产地一年生桔梗的总皂苷含量。结果表明，某些地区的一年生桔梗达到中国药典标准，可以人药，因而各产地应根据实际情况来决定桔梗的采收。     

天王补心丸全方及方中缺桔梗对小鼠镇静催眠作用的影响

目的:探讨中药古方天王补心丸对小鼠中枢神经系统的药理影响及舟楫之剂桔梗在该方剂中的作用.方法:采用小鼠自发活动计数、延长戊巴比妥钠睡眠时间及抗惊厥试验等方法,观察天王补心丸方、缺桔梗的天王补心丸方、天王补心丸中臣药酸枣仁、酸枣仁与桔梗配伍及其桔梗单独使用时的镇静、催眠与抗惊厥作用.结果:天王补心丸有良好的镇静、催眠与抗惊厥作用,但该方去掉桔梗后,镇静、催眠与抗惊厥效果明显下降;天王补心丸中重要成分酸枣仁本身有较好的镇静催眠作用加上桔梗后效果得到加强,但不如天王补心丸全方,桔梗本身没有镇静、催眠与抗惊厥作用.结论:在天王补心丸方剂的配伍中,桔梗作为舟楫之剂确实能载药上浮,增强该方对中枢神经系统的作用.     

Screening and appraisal for immunological adjuvant-active fractions from Platycodon grandiflorum total saponins

In this study, the total saponins from the root of Platycodon grandiflorum (PGS_t) was subjected to D101 macroreticular resin column chromatography to afford four fractions (PGS₃₀, PGS₅₀, PGS₇₅ and PGS₉₅). PGS_t and its four fractions were evaluated and compared for the haemolytic activities and adjuvant potentials on the specific cellular and humoral immune responses of ICR mice against recombinant hepatitis B surface antigen (HBsAg). PGS_t, PGS₃₀, PGS₅₀, PGS₇₅, and PGS₇₅ showed a slight haemolytic effect, with their concentration inducing 50% of the maximum haemolysis (HD₅₀) being 16.13?±?0.81, >200, 17.53?±?0.24, 20.16?±?0.76, 76.31?±?2.20?μg/mL against 0.5% rabbit red blood cell, respectively. PGS_t, PGS₅₀, and PGS₇₅ significantly not only enhanced the Con A-, lipopolysaccharide-, and HBsAg-induced splenocyte proliferation, but promoted the killing activities of natural killer (NK) cells from splenocytes in HBsAg-immunized mice (P?<?0.01 or P?<?0.001). HBsAg-specific IgG, IgG1, IgG2a, and IgG2b antibody levels in serum were also significantly enhanced by PGS_t, PGS₅₀, and PGS₇₅ compared with HBsAg control group (P?<?0.05, P?<?0.01, or P?<?0.001). Moreover, the adjuvant effects of PGS₅₀ and PGS₇₅ on the cellular immune responses and HBsAg-specific IgG2a and IgG2b antibody responses were more significant than those of Alum, PGS₃₀, and PGS₉₅. The results indicated that PGS₅₀ and PGS₇₅ could improve both cellular and humoral immune responses, and elicit a balanced Th1/Th2 response to HBsAg in mice, and that PGS₇₅ may be developed as an ideal candidate adjuvant for hepatitis B vaccine.     

正交设计考察不同干燥方法对桔梗质量的影响

目的确立桔梗的产地加工的干燥工艺。方法以药典方法并结合其他方法测定桔梗的出干率、总皂苷与水溶性浸出物的含量。结果桔梗采集后,洗净,以微波3min或80℃烘干较好,尤以微波干燥其出干率、桔梗总皂苷、水浸出物含量最高。结论通过微波干燥所得的桔梗饮片其桔梗总皂苷的含量明显高于晒干法,降低劳动强度,是一种值得研究和推广的干燥方法。     

Platycodon grandiflorum extract represses up-regulated adipocyte fatty acid binding protein triggered by a high fat feeding in obese rats

AIM： To investigate the effect of Platycodon grandiflorum extract （PGE） on lipid metabolism and FABP mRNA expression in subcutaneous adipose tissue of high fat diet-induced obese rats.
METHODS： PGE was treated to investigate the inhibitory effect on the pre-adipocyte 3T3-L1 differentiation and pancreatic lipase activity. Male Sprague-Dawley rats with an average weight of 439.03± 7.61 g were divided into four groups： the control groups that fed an experimental diet alone （C and H group） and PGE treatment groups that administered PGE along with a control diet or HFD at a concentration of 150 mg/kg body weight （C ＋ PGE and H ＋ PGE group, respectively） for 7 wk. Plasma total cholesterol （TC） and triglycerol （TG） concentrations were measured from the tail vein of rats. Adipocyte cell area was measured from subcutaneous adipose tissue and the fatty acid binding protein （FABP） mRNA expression was analyzed by northern blot analysis.
RESULTS： PGE treatment inhibited 3T3-L1 pre-adipocyte differentiation and fat accumulation, and also decreased pancreatic lipase activity. In this experiment, PGE significantly reduced plasma TC and TG concentrations as well as body weight and subcutaneous adipose tissue weight. PGE also significantly decreased the size of subcutaneous adipocytes. Furthermore, it significantly repressed the up-regulation of FABP mRNA expression induced by a high-fat feeding in subcutaneous adipose tissue.
CONCLUSION： PGE has a plasma lipid lowering-effect and anti-obesity effect in obese rats fed a high fat diet.From these results, we can suggest the possibility that PGE can be used as a food ingredient or drug component to therapeutically control obesity.     

Anti-inflammatory effects of YeongyoSeungma-tang

Ethnopharmacological relevance

YeongyoSeungma-tang which includes Fructus Forsythia, has been used in oriental traditional medicine for treatment of early smallpox and atopic dermatitis.

Aim of the study

YeongyoSeungma-tang was carried out to investigate for anti-inflammatory effects.

Materials and methods

YeongyoSeungma-tang was tested for anti-inflammatory effects against lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E₂ (PGE₂), and tumor necrosis factor alpha (TNF-α) releases as well as nuclear factor kappa B (NF-κB) expression using RAW264.7 macrophage cells.

Results

YeongyoSeungma-tang significantly inhibited generation of NO (42% and 59% inhibition at doses of 5 μg/mL and 10 μg/mL, respectively), PGE₂ (46% and 80% inhibition at doses of 5 μg/mL and 10 μg/mL, respectively) and TNF-α (6% and 23% inhibition at doses of 5 μg/mL and 10 μg/mL, respectively) on LPS-stimulated RAW 264.7 cells in a concentration-dependent manner. Consistently in these observations, the expression of inducible NO synthase (iNOS) enzyme was also inhibited by YeongyoSeungma-tang. However, YeongyoSeungma-tang did not show any influence on the expression of cyclooxygenase-2. The cream containing 0.075% YeongyoSeungma-tang showed good skin moisturizing effect without any irritation.

Conclusion

The present study may support the fact that YeongyoSeungma-tang can have the good possibility as an anti-inflammatory agent for troubled skins.     

固相萃取气相色谱内标法测定桔梗中拟除虫菊酯的农药残留

 目的 本实验建立了桔梗中甲氰菊酯、高效氯氟氰菊酯、溴氰菊酯、氰戊菊酯、氯菊酯、联苯菊酯等 6 种拟除虫菊酯类农药残留的提取、净化及其气相色谱法。 方法 样品运用石油醚提取,乙腈 - 二氯甲烷 (5:95) 洗脱, GC-ECD 检测 , 内标法定量。 结果 6 种拟除虫菊酯农药回收率在 78.86% ～ 97.80% , RSD 在 1.17% ～ 11.63% ,可以满足农药残留分析的要求。 结论 方法灵敏度高,选择性强,操作简单、快速,净化效果好等特点,可应用于桔梗药材中痕量农药残留的检测。     

桔梗与氯丙嗪伍用对大鼠脑纹状体多巴胺的影响

目的：研究桔梗与氯丙嗪联合应用对大鼠脑纹状体单胺类神经递质多巴胺的影响,探讨中西药配伍应用后可能发生的药物相互作用与机制。方法：大鼠随机分成4组：生理盐水组、桔梗(生药15.2 g·kg^-1·d^-1)单独给药组、氯丙嗪(20 mg·kg^-1·d^-1)单独给药组、氯丙嗪与桔梗(20 mg·kg^-1·d^-1+15.2 g·kg^-1·d^-1)合并给药组,每组6只,给药10 d后,利用微透析法在大鼠脑纹状体取样,HPLC仪检测透析液中多巴胺水平。结果：药后140 min(取样点80 min)时,氯丙嗪＋桔梗组中多巴胺水平(1.52±0.34) μg·L^-1显著高于氯丙嗪组(1.25±0.22) μg·L^-1(P<0.05)和生理盐水组(1.06±0.24) μg·L^-1(P<0.01)。结论：桔梗与氯丙嗪合用后大鼠脑内多巴胺水平升高,治疗时同时给予桔梗,可以减少氯丙嗪的用量,达到保持疗效,降低其不良反应。