【作者】 马容；

【导师】 狄留庆；

【作者基本信息】 南京中医药大学 ， 药剂学， 2008， 硕士

【摘要】 山茱萸为山茱萸科植物山茱萸的干燥成熟果肉,于秋末冬初果皮变红时进行采收,用文火烘或置沸水中略烫后及时除去果核,干燥而得。山茱萸味酸、涩、微温,归肝、肾经,具有补益肝肾,涩精固脱之功效。它既能平补,又能固脱;既能补阴,又能壮阳,而且药性平和,因此用途广泛。本文对山茱萸防治糖尿病肾病的有效部位进行了研究。文献综述研究,包括:山茱萸化学成分研究进展,山茱萸药理作用研究进展,山茱萸环烯醚萜总苷的研究进展。山茱萸多糖的研究进展。山茱萸防治糖尿病肾病有效部位的研究,包括:1、山茱萸药材品质评价及分析方法的建立。采用HPLC法测定了山茱萸药材中马钱苷和莫诺苷的含量。结果显示,马钱苷的含量达到7.8mg·g^-1,莫诺苷的含量达到10.8mg·g^-1。符合药典“本品按干燥品计算,含马钱苷不得少于0.60%”的要求。此外,采用硫酸蒽酮法,测定了山茱萸中总多糖的含量,结果显示,山茱萸中总多糖的平均含量为2.33%。2、山茱萸中有效部位环烯醚萜总苷的提取。采用正交试验法,以马钱苷、莫诺苷的提取率为指标,优选出最佳的醇提工艺为:山茱萸药材用6倍量80%的乙醇提取2次,每次1小时。利用HPD100型大孔吸附树脂对山茱萸总苷进行精制,考察了树脂的吸附容量,乙醇的洗脱浓度以及乙醇洗脱体积,从而确定了最佳工艺为:醇提液浓缩水沉,将上清液加水定容至0.5g生药·mL^-1,此提取液上大孔树脂进一步分离纯化,上样后,用30%的乙醇,12倍量进行洗脱,洗脱液浓缩,减压干燥至恒重,得到山茱萸环烯醚萜总苷。3、山茱萸总多糖的提取。通过正交试验法,采用硫酸蒽酮法测定多糖的含量,优选出多糖的最佳水提工艺为:山茱萸药材用12倍量水煎煮2次。每次2小时。进一步考察了多糖的醇沉工艺,得到其最佳工艺为:水提液浓缩至0.5g·mL^-1,加95%的乙醇醇沉至80%。4、山茱萸防治糖尿病肾病活性部位群的筛选。运用中药血清药理学方法研究山茱萸活性部位群对高糖致RMC增殖的体外抑制作用,并与总提取物组进行比较,优选山茱萸防治糖尿病肾病的最佳活性部位群组合,采用四甲基氮唑盐（MTT）比色法。结果显示,山茱萸各有效部位合理组合后,具有不同程度的对抗高糖致RMC的增殖作用,并呈一定程度的量效关系,初步确定山茱萸总苷配伍多糖为防治糖尿病肾病的最佳活性部位群。另外,还通过对小鼠DN模型整体筛选,研究了山茱萸防治糖尿病肾病的药效物质基础,确定有效部位在药效靶点上的作用,追踪活性部位（群）。分离、纯化山茱萸中环烯醚萜总苷、多糖、三萜酸,以环烯醚萜总苷为基础将各有效部位进行合理组合,以DN模型下的降血糖、降血清尿素氮活性及降低肾指数为指标,并与总提取物组进行比较,筛选、确定山茱萸防治DN的有效部位群。结果显示,山茱萸各有效部位合理组合后,均能延缓小鼠肾病变的进程及降低肾病变的严重程度,其中以山茱萸环烯醚萜总苷+多糖小剂量组效果最佳。所以,环烯醚萜总苷+多糖为山茱萸防治DN的最佳活性部位群。5、在体肠循环灌流法考察山茱萸中环烯醚萜总苷在大鼠小肠的吸收特征,包括:（1）马钱苷、莫诺苷在空白肠灌流液中的稳定性。结果显示,两者在空白肠灌流液中孵育后的浓度为孵育前的98.56%、99.56%,两者在肠循环液中比较稳定。（2）肠壁对马钱苷、莫诺苷物理吸附的影响。结果显示,经4h孵育后,马钱苷浓度为原来浓度的（99.16±0.23）%（n=3）,莫诺苷浓度为原来浓度的（95.01±0.21）%（n=3）,可以认为大鼠小肠壁对药物基本没有物理吸附。（3）不同肠段对大鼠小肠吸收的差异。结果显示,回肠与十二指肠、回肠与空肠比较P＜0.05,差异有统计学意义,表明在回肠中,两者吸收较少,而十二指肠与空肠的P＞0.05,差异无统计学意义,表明两者在十二指肠和空肠的吸收速率相近。（4）不同浓度对大鼠小肠吸收的影响。结果显示,在0.5～2 mg·mL^-1内,随着浓度的升高,马钱苷和莫诺苷的Ka略有增大,但无显著差异,表明在此浓度范围内,两者的吸收符合线性动力学特征。更多还原

【Abstract】 Cornus officinalis is widely used in traditional chinese medicine.This thesis studied the chemical compounds and the intestinal absorption characteristics of Cornus officinalis.Literature investigations of Cornus officinalis were consulted,including:progress in research of the active chemical components of Cornus officinalis,progress in studies on pharmacological effect of Cornus officinalis,research advance of total iridoid glycoside in Cornus officinalis,studies on the polysaccharides from Cornus officinalis.Investigation of the chemical compounds and the intestinal absorption characteristics of Cornus officinalis,including:1.To establish the quality evaluation and analysis methods of Cornus officinalis.To determine the content of loganin and morroniside by high performance liquid chromatography.The results showed that the content of loganin was up to 7.8mg·g^-1,the content of morroniside was up to 10.8mg·g^-1.Chinese pharmacopoeia demands that the content of loganin shouldn’t be lower than 0.60%.The medicinal materials reached the requirements and can be used in experiments.The anthrone-sulfuric method was used to determine the content of polysaccharides from Cornus officinalis.The results showed that the content of polysaccharides from Cornus officinalis was up to 2.33%.2.Studies on the extraction of the total iridoid glycoside in Cornus officinalis.The orthogonal experimental method was used by choosing the extraction rates of loganin and morroniside as index to decide the best alcohol extraction process.The best process was: adding 6 times amount of 80%ethanol,reflux and extract for 1 hour,2 times totally. HPD100 macroporous adsorption resin was used to purify the total iridoid glycoside in Cornus officinalis.The saturation capability of adsorption,the elution concentration of ethanol and the elution volume were investigated.The best process was:the alcohol extract was concentrated and extracted with water,the supernatant was concentrated to 0.5g·mL^-1. HPD100 macroporous adsorption resin was used to separate and purify this supernatant. The best elution concentration of ethanol was 30%and the elution volume was 12 times.The eluate was concentrated and vacuum dried to the constant weight.The solids were the total iridoid glycoside. 3.Studies on the extraction of the polysaccharides in Cornus officinalis.The orthogonal experimental method and anthrone-sulfuric method were used.The best process was:adding 12 times amount of water,reflux and extract for 2 hour,2 times totally.The best water extracting and alcohol precipitation was:the water extract was concentrated to proportion as 0.5g·mL^-1（drug materials/solution）,95%alcohol was added to the solution to 80%alcohol.4.To screen its optimum component group in preventing diabetic nephropathy through the research on the effect of different component groups of Cornus officinalis medcinal serum on inhibiting the proliferation of glomerular mesangial cells cultured in high glucose medium.The proliferation of RMC is analyzed by MTT method（490nm）.The results showed that all kinds of component groups inhibit the proliferation of GMC induced by high glucose medium in different degree and present obvious relationship of quantity-effect. （HXMTZG+DT） is confirmed preliminarily as the optimum component group in preventing diabetic nephropathy.To study the material foundation of the effectiveness of Cornus officinalis for preventing diabetic nephropathy,determine the functions of its effectiveness and trace its components.To separate and purify the components of the said drug,e.i.,HXMTZG,organic acid and amylose and screening and determine its effectiveness components for diabetic nephropathy by taking for indexes the results of the experiment of decreasing blood glucose,BUN and kidney index,taking HXMTZG as the basic content and reasonably combining screening of all the effective components.The results showed that all kinds of component groups inhibit the progress of the diabetic nephropathy and alleviate the damage of kidney,but the action of（HXMTZG+DT） group is the best.（HXMTZG+DT） is confirmed finally as the optimum component group in preventing diabetic nephropathy.5.The intestinal absorption characteristics of loganin and morroniside were investigated with the model in situ intestinal absorption in rats.（1） Stability of loganin and morroniside in blank intestinal perfusate after incubation. The results were:98.56%of loganin;99.56%of morroniside.They all fit the detection of biological specimen.（2） After incubation for 4 hour in loganin and morroniside,the concentration of loganin was 99.16%,the concentration of morroniside was 95.16%.It can be concluded that the intestinal has no effect on absorption.（3） Study on the absorption of loganin and morroniside at different segments of intestine.There were no significant differents of the absorption in duodenum and in jejunum. The absorption in duodenum and in jejunum were faster than in ileum.（4） Within 0.5～2 mg·mL^-1,the absorption rate constants Ka were not changed much by concentrations.The absorption of loganin and morroniside fit linear dynamics.The relationship of the absorption amount and dose was consistent with direce ratio.The absorption mechanism of these compounts was passive diffusion.更多还原