自拟更年汤对肾阳虚型围绝经期模型大鼠雌激素样作用的研究

目的探讨自拟更年汤对肾阳虚型围绝经期模型大鼠的雌激素样作用机制。方法选择阴道细胞学表现为有规律的4~5 d动情周期的4月龄雌性SD大鼠20只为青年组,阴道细胞学表现无动情周期改变的18月龄雌性SD大鼠20只为老年组;选用自然老化的围绝经期雌性SD大鼠,肌注氢化可的松构建肾阳虚型围绝经期大鼠模型,将造模成功大鼠随机分为更年汤组、教材方组、利维爱组和生理盐水组。青年组、老年组、生理盐水组均给予生理盐水灌胃,各给药组给予相应药物灌胃,灌胃15 d后处死大鼠,分离摘取子宫称子宫湿质量,计算子宫指数,采用免疫组化及原位杂交检测方法观察大鼠子宫组织中血管内皮生长因子(VEGF)及其信使VEGF mRNA的阳性表达情况。结果各给药组子宫湿质量、子宫指数均明显高于生理盐水组和老年组(P均0.05);更年汤组子宫湿质量、子宫指数与青年组比较差异均无统计学意义(P均0.05),与利维爱组、教材方组比较差异均有统计学意义(P均0.05);利维爱组子宫湿质量、子宫指数与教材方组比较差异均无统计学意义(P均0.05)。老年组和生理盐水组VEGF及VEGF mRNA阳性表达明显低于青年组(P均0.05)。各给药组VEGF及VEGF mRNA的阳性表达明显高于生理盐水组、老年组(P均0.05),更年汤组、利维爱组VEGF及VEGF mRNA阳性表达明显高于教材方组(P均0.05),利维爱组VEGF及VEGF mRNA阳性表达与更年汤组比较差异均无统计学意义(P均0.05)。结论自拟更年汤可促进子宫增生质量,具有雌激素样作用;其促进子宫增加质量的机制可能与上调子宫组织中VEGF及VEGFmRNA的阳性表达有关。     

自拟更年汤对肾阳虚型围绝经期模型大鼠下丘脑雌激素受体及肾上腺皮质Bax、Bcl-2蛋白表达的影响

目的探讨自拟更年汤治疗围绝经期综合征肾阳虚证的可能作用机制。方法选取11~12月龄动情周期紊乱、动情间期延长的雌性大鼠,并予氢化可的松注射液0.5ml/100 g连续肌注9天,建立肾阳虚型围绝经期综合征大鼠模型。将造模成功大鼠随机分为更年汤组、右归丸组、替勃龙组和模型组,每组20只。更年汤组大鼠给予自拟更年汤溶液18 g/(kg·d)灌胃,右归丸组给予右归丸溶液16 g/(kg·d)灌胃,替勃龙组给予替勃龙片溶液0.294 mg/(kg·d)灌胃。另取4月龄大鼠(青年组)20只,18月龄大鼠(老年组)20只,模型组、青年组、老年组大鼠用生理盐水4ml灌胃,各组连续灌胃15天。检测各组大鼠下丘脑雌激素受体(ER)和ER mRNA表达及肾上腺皮质Bax、Bcl-2蛋白表达。结果更年汤组与右归丸组、模型组、青年组下丘脑ER阳性表达比较差异均有统计学意义(P0.05或P0.01);各组大鼠下丘脑ER mRNA阳性表达均无统计学意义(P0.05)。模型组肾上腺球状带Bax蛋白表达显著增加,而Bcl-2蛋白明显降低,自拟更年汤可下调Bax蛋白表达、上调Bcl-2蛋白表达(P0.05);在束状带,自拟更年汤亦可下调Bax蛋白表达(P0.05),而对Bcl-2蛋白表达影响不明显(P0.05);在网状带,自拟更年汤对Bax蛋白表达具有降低作用,对Bcl-2蛋白明显上调,优于右归丸组和替勃龙组(P0.05)。结论自拟更年汤治疗围绝经期综合征肾阳虚证可能与提高下丘脑ER、肾上腺皮质Bcl-2蛋白表达及降低肾上腺皮质Bax蛋白表达水平有关。     

更年汤对围绝经期模型大鼠雌激素受体及细胞凋亡相关因子的影响及作用机理研究

目的:探讨更年汤对围绝经期模型大鼠下丘脑ER及卵巢颗粒细胞凋亡因子Bax及BaxmRNA的影响。方法:将雌性11-14月龄SD大鼠经阴道脱落细胞筛选,符合自然老化型围绝经期大鼠模型要求的大鼠随机分为生理盐水组、教材方组、更年汤组,另选4月龄雌性大鼠为青年对照组,18月龄雌性大鼠为老年对照组。相应药物灌胃成功后,采用病理切片了解大鼠卵巢形态学变化,免疫组化及原位杂交检测方法观察大鼠下丘脑ER、Bax及BaxmRNA的阳性表达情况。结果:生理盐水组的ER阳性表达与教材方组、更年汤组比较均有统计学意义(P<0.05),更年汤组与教材方组ER阳性表达比较无统计学意义(P>0.05);更年汤组与生理盐水组比较,卵泡发育、闭锁及扩张观察有统计学意义或有显著统计学意义(P<0.05或P<0.01);与教材方组比较,卵泡发育及闭锁观察比较有显著统计学意义或有统计学意义(P<0.01或P<0.05),卵泡扩张观察无统计学意义(P>0.05);更年汤组的Bax及BaxmRNA阳性表达与生理盐水组及教材方组的比较均有统计学意义(P<0.05)。结论:更年汤能改善围绝经期模型大鼠下丘脑ER阳性表达,有促进卵巢内卵泡发育,抑制卵巢颗粒细胞凋亡因子Bax及BaxmRNA阳性表达的作用,并且作用较教材方显著。     

自拟更年汤对肾阳虚型围绝经期模型大鼠卵巢细胞凋亡相关因子的影响

目的探讨自拟更年汤对肾阳虚型围绝经期模型大鼠卵巢组织形态学及卵巢颗粒细胞凋亡因子bax、bax mRNA、bcl-2、bcl-2 mRNA的影响。方法选用自然老化的围绝经期雌性SD大鼠,臀部肌肉左右交替肌注氢化可的松构建肾阳虚型围绝经期大鼠模型,将造模成功大鼠随机分为更年汤组、教材方组、利维爱组、生理盐水组;另选择4月龄SD雌性大鼠20只作为青年组,18月龄SD雌性大鼠20只作为老年组。青年组、老年组、生理盐水组均给予生理盐水灌胃,各给药组给予相应药物灌胃,灌胃15 d后处死大鼠,分离摘取双侧卵巢,运用病理切片观察卵巢组织大体形态,运用免疫组化及原位杂交法检测卵巢颗粒细胞凋亡因子bax、bax mRNA、bcl-2、bcl-2 mRNA表达情况。结果各给药组卵泡发育良好情况优于老年组和生理盐水组(P均0.05),卵泡闭锁、卵泡扩张情况轻于老年组和生理盐水组(P均0.05);自拟更年汤组与利维爱组卵泡发育良好情况优于教材方组(P均0.05),自拟更年汤组与利维爱组比较差异无统计学意义(P0.05);自拟更年汤组卵泡闭锁、卵泡扩张情况轻于教材方组和利维爱组(P均0.05),教材方组与利维爱组比较差异均无统计学意义(P均0.05)。各给药组bax、bax mRNA阳性表达明显低于老年组和生理盐水组(P均0.05),bcl-2与bcl-2 mRNA阳性表达明显高于老年组和生理盐水组(P均0.05);自拟更年汤组与利维爱组bax、bax mRNA阳性表达明显低于教材方组(P均0.05),bcl-2与bcl-2 mRNA阳性表达明显高于教材方组(P均0.05),自拟更年汤组bax、bax mRNA、bcl-2、bcl-2 mRNA阳性表达与利维爱组比较差异无统计学意义(P均0.05)。结论自拟更年汤能促进卵泡发育,抑制卵泡闭锁和扩张,能上调卵巢颗粒细胞抑制凋亡因子bcl-2、bcl-2 mRNA的阳性表达,抑制促凋亡因子bax、bax mRNA的阳性表达。     

更年汤对肾阳虚型围绝经期模型大鼠IL-6及骨密度影响的实验研究

目的:观察更年汤对肾阳虚型围绝经期模型大鼠白细胞介素(IL)-6及骨密度的影响,探讨其机制。方法:80只11~12月龄自然围绝经期SD大鼠为研究对象,随机分为更年汤组、教材方组、利维爱组、模型组,每组20只;4月龄、18月龄雌性大鼠分别作为对照组,每组各20只。结果:与青年组比较,模型组、老年组、更年汤组、教材方组、利维爱组的血清IL-6含量较高,差异有统计学意义(P0.05);与模型组比较,更年汤组、教材方组、利维爱组的血清IL-6含量较低(P0.05),老年组的血清IL-6含量较高(P0.05)。各组大鼠骨密度比较,差异有统计学意义(P0.05),模型组较青年组骨密度有下降(P0.05),模型组与更年汤组、教材方组、利维爱组比较,差异也有统计学意义(P0.05)。利维爱组与教材汤组比较,差异有统计学意义(P0.05),与更年汤组比较,差异无统计学意义(P0.05)。更年汤组与教材汤比较,差异有统计学意义(P0.05)。结论:更年汤能降低肾阳虚型模型大鼠血清IL-6含量,提高骨密度水平。     

自拟更年汤对肾阳虚围绝经期模型大鼠卵巢功能的影响

目的:探讨自拟更年汤对肾阳虚围绝经期模型大鼠卵巢功能的影响。方法:选取10～12个月龄雌性SD大鼠,采集阴道脱落细胞筛选出围绝经期模型大鼠,将围绝经期模型大鼠随机分为模型组、更年汤低剂量组、更年汤中剂量组、更年汤高剂量组、替勃龙组,每组24只,构建肾阳虚模型;另选24只4～6个月龄的青年大鼠为青年组;各组分别给予相应药物灌胃,末次给药24 h后,检测血清中卵泡刺激素(FSH)水平及卵巢组织中FSHR、IGF-1以及其受体蛋白表达。结果:更年汤高剂量组、更年汤中剂量组、替勃龙组大鼠血清FSH水平明显低于模型组(P0.05或P0.01),卵巢组织中FSHR、IGF-1及其受体表达明显高于模型组(P0.05或P0.01)。结论:更年汤能改善肾阳虚围绝经期模型大鼠卵巢功能,可能与调节大鼠血清中FSH水平,调节大鼠卵巢组织中FSH、IGF-1及其受体的表达有关。     

补肾中药对去卵巢大鼠骨组织ER、OPG表达的促进作用

通过检测去卵巢大鼠给予补肾中药后腰椎内ERmRNA及OPG mRNA的表达,进一步探讨补肾中药防治骨质疏松的分子机制.将雌性3月龄SD大鼠48只,随机分为卵巢切除 补肾中药防治(HDP),卵巢切除 雌激素(倍美力)防治(ER),骨质疏松(卵巢切除)(OVX)及正常对照组(SHAM)4组,每组12只.术后12周处死大鼠,取L3椎体,异硫氰酸胍一步法提取总RNA,嵌套式反转录聚合酶链反应(NRT-PCR),扩增待检测各组ERmRNA及OPGmRNA的表达.同时取L4椎体,行骨组织形态计量学检测.结果显示HDP组ER mRNA及OPG mRNA阳性表达率分别为75.0%和66.7%;EP组ER mRNA及OPG mRNA阳性表达率均为83.3%;OVX组各例ER mRNA均无表达,2例OPG mRNA呈弱阳性表达(阳性率为16.7%);SHAM组呈ER mRNA及OPG mRNA阳性表达.χ2检验,HDP组与ER组比较ER mRNA及OPG mRNA阳性表达率均无显著性差异(P＞0.05);该两组与OVX组比较,ER mRNA及OPG mRNA阳性表达率均有高度显著性差异(P＜0.01);而与SHAM组比较,HDP组ERmRNA,ER组ER mRNA及OPG mRNA阳性表达率无显著性差异(P＞0.05),但HDP组OPG mRNA阳性表达率有显著性差异(0.01＜P＜0.05).骨组织形态计量学检测,OVX组骨体积、骨小梁厚度及骨小梁数目均比SHAM组显著减少,而骨小梁间隔显著增大.同EP组相似,HDP组骨体积、骨小梁厚度显著增加,虽也使骨小梁间隔稍有减少,但与OVX组比较无统计学差异.表明补肾中药通过直接作用于骨组织中ER,增强ER mRNA的表达,并刺激OPG的分泌,从而有望替代雌激素,对去卵巢大鼠的骨质疏松有明显的防治作用.     

更年汤对肾阳虚围绝经期模型大鼠肾上腺皮质中Bax和Bcl-2蛋白表达的影响

目的:探讨中药复方更年汤对肾阳虚型围绝经期模型大鼠肾上腺皮质中Bax、Bcl-2表达的影响。方法:建立自然衰老的雌性肾阳虚型围绝经期大鼠模型,11~12月龄60只,随机分为更年汤组、利维爱组、生理盐水组,每组20只;4月龄20只作为青年组;18月龄20只作为老年组。用免疫组化法检测肾上腺皮质Bax、Bcl-2的表达。结果:与生理盐数组比较,利维爱组肾上腺皮质Bax表达降低、Bcl-2的表达升高,差异均有统计学意义(P0.05)。更年汤组肾上腺皮质Bax阳性总积分表达低于利维爱组、生理盐水组,Bcl-2阳性总积分表达高于利维爱组、生理盐水组,差异均有统计学意义(P0.05),更年汤组稍优于利维爱组。结论:自拟更年汤能降低肾阳虚型围绝经期模型大鼠肾上腺皮质Bax的表达,提高肾上腺皮质Bcl-2表达水平,平衡围绝经期内分泌失调。     

更年汤对围绝经期模型大鼠E_2、LH、FSH、GnRH的干预作用研究

目的:通过构建围绝经期大鼠模型,观察更年汤对模型大鼠内分泌相关指标的影响,探讨更年汤延缓卵巢衰老的作用机制。方法:将SD雌性大鼠随机分为更年汤组、教材方组、模型组、青年组、老年组各10只。前三组采用自然老化的围绝经期大鼠模型,青年和老年组为正常青年、老年模型。分别灌胃更年汤、教材方和生理盐水15日,于末次给药24h后麻醉,颈动脉取血5mL,剖取子宫、卵巢,观察血清卵泡刺激素(FSH)、黄体生成激素(LH)、雌二醇(E2)、促性腺激素释放激素(GnRH)和子宫、卵巢指数。结果:与模型组比较,治疗组、对照组E2含量均升高,治疗组有非常显著性差异(P<0.01);治疗组、对照组均能显著降低血清LH含量(P<0.01),组间比较有非常显著性差异(P<0.01);治疗组、对照组FSH含量均显著降低(P<0.01)组间比较无显著性差异(P>0.05);治疗组能显著降低血清GnRH含量(P<0.01),组间比较有非常显著性差异(P<0.01);治疗组、对照组均使卵巢、子宫指数升高,有非常显著性差异(P<0.01),卵巢组间比较差异无显著性差异(P>0.05),子宫组间比较有显著性差异(P<0.05)。结论:更年汤可明显提高模型大鼠E2水平,升高大鼠卵巢、子宫指数,降低血清FSH、LH、GnRH水平,从而达到延缓卵巢功能衰退的作用。     

更年汤对肾阳虚围绝经期模型大鼠肾上腺皮质中Bax和Bcl-2蛋白表达的影响简

目的:探讨中药复方更年汤对肾阳虚型围绝经期模型大鼠肾上腺皮质中Bax、Bcl-2表达的影响。方法:建立自然衰老的雌性肾阳虚型围绝经期大鼠模型,11~12月龄60只,随机分为更年汤组、利维爱组、生理盐水组,每组20只;4月龄20只作为青年组;18月龄20只作为老年组。用免疫组化法检测肾上腺皮质Bax、Bcl-2的表达。结果:与生理盐数组比较,利维爱组肾上腺皮质Bax表达降低、Bcl-2的表达升高,差异均有统计学意义(P<0.05)。更年汤组肾上腺皮质Bax阳性总积分表达低于利维爱组、生理盐水组,Bcl-2阳性总积分表达高于利维爱组、生理盐水组,差异均有统计学意义(P<0.05),更年汤组稍优于利维爱组。结论:自拟更年汤能降低肾阳虚型围绝经期模型大鼠肾上腺皮质Bax的表达,提高肾上腺皮质Bcl-2表达水平,平衡围绝经期内分泌失调。     

Isolation and hypoglycemic activity of eleutherans A, B, C, D, E, F, and G: glycans of Eleutherococcus senticosus roots

Intraperitoneal injection of an aqueous extract of the crude drug "shigoka" (Siberian ginseng), Eleutherococcus senticosus roots, remarkably diminished plasma-sugar level in mice. Fractionation of the extract by monitoring the activity yielded seven glycans, eleutherans A, B, C, D, E, F, and G, which exerted marked hypoglycemic effects in normal and alloxan-induced hyperglycemic mice.     

Phytochemistry, Metabolism, and Metabolomics of Ginseng

Ginseng, as a medicinal plant, has been used for thousands of years in China, Korea, and Japan, and the study on ginseng is a hotspot in the research field as evidenced by about 7000 scientific papers in PUBMED. In recent decades, many ginseng studies focused on the metabolism and metabolomics of ginseng or its active ingredients using modern bioanalytical technologies. To date, more than 200 ginsenosides and non-saponin constituents have been isolated and identified. In the past decades, rapid development of analytical technologies has facilitated the advancement of ginseng research in many ways. In this review, we focus on the advances of ginseng research in chemistry, pharmacology, and metabolomics. We also provide the comments on the significance as well as challenges of metabolomics-based ginseng studies.     

Correlation of Metabolism, Metabolomics, and Metabonomics

<正>In this issue,CHM published two reviews on drug metabolism of Chinese herbal medicines.Three key words have been found,they are metabolism,metabolomics,and metabonomics.As we known,drug metabolism is the biochemical modification of pharmaceutical substances respectively by living organisms through specialized enzymatic systems,the rate of metabolism determines the duration and intensity of pharmacological responses     

Antimicrobial, anti-inflammatory, antiparasitic, and cytotoxic activities of Galium mexicanum

Ethnopharmacological relevance

To study the potential benefit of the traditional Mexican medicinal plant Galium mexicanum Kunth (Rubiaceae). Hexane, chloroform, and methanol extracts as well as various fractions from these extracts were tested to determine antibacterial, antifungal, antiparasitic or anti-inflammatory activities in vitro.

Materials and methods

Aerial parts of the plant were extracted with various solvents and fractionated accordingly. Their antibacterial and antifungal activities were assessed on nine bacterial and four fungal strains. Leishmania donovani was used as a protozoan strain for antiparasitic activity. The anti-inflammatory activity of the compounds was investigated by measuring the secretion of interleukin-6 when macrophages were exposed to lipopolysaccharide.

Results

Various extracts and fractions obtained from this plant exhibit antibacterial, antifungal, antiparasitic, and anti-inflammatory activities. Of special interest was the hexane fraction HE 14b, which show antibacterial (ranging between 67 and 666 μg/ml) and antifungal (at concentrations of 333 μg/ml) activities. Also the hexane fraction HE 5 exhibited antiparasitic activity (at concentrations of 260 μg/ml), whereas the methanol fraction ME 13-15 showed a potent anti-inflammatory activity when compared to dexamethasone. Chemical analyses of the chloroform extract show the presence of triterpenes, saponins, flavonoids, sesquiterpene lactones, and glucosides, but no tannins were detected in the assayed extract.

Conclusions

The benefit of Galium mexicanum as a traditional medicinal plant was confirmed using antibacterial and antifungal assays in vitro. We also report for the first time, and to the best of our knowledge, antiparasitic and anti-inflammatory activities of this plant.     

Isolation and structure of homotemsirolimuses A, B, and C

Homotemsirolimuses A, B, and C (2a, 2b, 2c) were found to be minor components of a temsirolimus preparation made from rapamycin. These three temsirolimus analogues are derived from the corresponding rapamycin analogues, homorapamycins A, B, and C (1a, 1b, 1c) produced by the strain Streptomyces hygroscopicus. The structures of homotemsirolimuses A, B, and C were determined by spectroscopic methods. These compounds were tested for mTOR kinase inhibition and in two proliferation assays using LNCap prostate and MDA468 breast cancer cells. The results suggested that the mTOR inhibition and antiproliferation potencies for 2a, 2b, and 2c are comparable to those of rapamycin (1) and temsirolimus (2).     

Antimalarial activity of physalins B, D, F, and G

The antimalarial activities of physalins B, D, F, and G (1-4), isolated from Physalis angulata, were investigated. In silico analysis using the similarity ensemble approach (SEA) database predicted the antimalarial activity of each of these compounds, which were shown using an in vitro assay against Plasmodium falciparum. However, treatment of P. berghei-infected mice with 3 increased parasitemia levels and mortality, whereas treatment with 2 was protective, causing a parasitemia reduction and a delay in mortality in P. berghei-infected mice. The exacerbation of in vivo infection by treatment with 3 is probably due to its potent immunosuppressive activity, which is not evident for 2.     

酸柏栀油软胶囊对小鼠学习记忆及脑中NO、Ach的影响

目的:观察酸柏栀油软胶囊对小鼠的促学习记忆作用并初步探讨其机制。方法:采用Morris水迷宫法观察酸柏栀油软胶囊对小鼠的学习记忆影响,并测定小鼠脑内NOS、ChaE、AChE的含量。结果:酸柏栀油软胶囊1.25～5 ml/kg能缩短小鼠登台潜伏期,增加穿越原平台次数,以中剂量作用最强;且登台潜伏期随训练时间的延长,各组作用均逐渐增强,d20的作用尤为明显。酸柏栀油软胶囊尚能增加小鼠脑内NOS、ChaE活性,降低AChE活性,也以中剂量作用为佳。结论:酸柏栀油软胶囊对小鼠学习记忆有促进作用,该作用可能与其富含不饱和脂肪酸,增加脑内NO、Ach的含量有关。     

鲜姜黄挥发油化学成分的GC-MS分析

目的:分析广西产鲜姜黄挥发油的化学成分,为其利用提供科学依据。方法:采用水蒸气蒸馏法提取鲜姜黄挥发油,用GC毛细管柱进行分析,归一化法测定其相对含量,并用GC-MS法鉴定化学成分。结果:检出48个色谱峰,鉴定出48个化合物,占总挥发油的100%。结论:广西产鲜姜黄油的的主要成分是姜烯(16.25%),1,8-桉叶油素(13.52%),芳-姜黄酮(13.55%),姜黄酮(11.19%),β-倍半水芹烯(10.08%),表-α-绿叶烯(6.26%),芳-姜黄烯(5.43%),大根香叶酮(5.02%),γ-松油烯(2.38%),红没药醇(3.59%)。