刺五加经发酵炮制后对小鼠抗应激作用的对比研究

目的:探讨刺五加经发酵炮制后对小鼠抗应激作用。方法:采用常压耐缺氧试验、低温冷冻试验、负重游泳试验及耐高温试验等实验方法,以小鼠存活时间为指标,开展刺五加经发酵炮制后对小鼠在冷、热、疲劳、缺氧等应激环境下的生存影响的实验研究。结果:与刺五加组比较刺五加发酵组可延长缺氧存活时间、低温存活时间、高温存活时间及游泳持续时间,其中延长游泳持续时间具有极显著性差异(P0.01)、缺氧存活时间、低温存活时间具有显著性差异(P0.05),高温存活时间虽有延长但无统计意义。结论:刺五加经发酵炮制后可延长小鼠在冷、热、疲劳、缺氧等应激环境下的生存时间。     

复方刺五加抗疲劳作用的实验研究

目的:观察复方刺五加对运动疲劳小鼠的影响.方法:以游泳小鼠为模型,观察小鼠力竭游泳时间;测定BLA、BUN等指标.结果:复方刺五加能不同程度增加小鼠力竭游泳时间,降低BLA、BUN值.结论:复方刺五加可有效改善小鼠运动疲劳状态.     

抗疲劳中药刺五加中1个新倍半萜

目的研究中药刺五加Acanthopanax senticosus的抗疲劳活性成分。方法采用硅胶、Sephadex LH-20、MCI gel CHP-20P、ODS等多种色谱技术分离纯化,通过质谱及核磁共振等数据对化合物进行结构鉴定;并采用ABTS法研究其体外清除自由基活性。结果从刺五加甲醇提取物中分离得到1个倍半萜类化合物7α(H)-桉烷-4α,5β,11,12-四醇-3-酮(1),该化合物自由基清除的半数清除浓度为(43.1±1.2)μg/m L。结论化合物1为1个新的eudesmane型倍半萜,命名为刺五加醇A;也是首次从五加科植物中分离得到此类化合物。     

刺五加水提物对小鼠抗疲劳及抗缺氧能力的影响

目的:观察刺五加水提物对小鼠的抗疲劳及抗缺氧作用。方法:将小鼠随机分为空白对照组,刺五加水提物高、中、低剂量组(30、20、10g/kg),连续灌胃14d后,测定小鼠负重游泳时间、常压缺氧条件下存活时间,并检测疲劳运动小鼠血乳酸水平。结果:刺五加水提物各剂量组均能延长小鼠的常压缺氧条件下存活时间和负重游泳时间,降低小鼠血乳酸水平,其中高、中剂量组作用尤为显著,与空白对照组的差异具有统计学意义(P0.05)。结论:刺五加水提物可显著提高小鼠抗疲劳及抗缺氧能力。     

刺五加抗疲劳活性部位中刺五加苷B的含量测定

目的：建立刺五加抗疲劳活性部位中刺五加苷B的含量测定方法。方法：用小鼠负重游泳实验，确定刺五加粗提物的抗疲劳活性部位；硅胶柱色谱，Sephadex LH-20柱色谱，制备液相色谱对活性部位分离纯化，并对分离所得化合物进行理化性质研究和结构鉴定；含量测定中高效液相色谱条件为Diamonsil C18柱(4.6 mm×250 mm,5 μm)，流动相乙腈-水-醋酸(10∶90∶0.01)，流速1.0 mL·min-1，紫外检测波长344 nm。结果：从刺五加抗疲劳活性部位As1中分离得到一化合物，鉴定为刺五加苷B；刺五加苷B的线性范围为 0.104～20.8 μg，r=0.999 9；平均回收率为97.68%，RSD 1.4%(n=6)。结论：从刺五加抗疲劳活性部位As1中分离得到含量较高的刺五加苷B，作为质量控制的指标成分，并建立了刺五加抗疲劳活性部位中刺五加苷B含量测定方法。     

基于网络药理学的刺五加总苷抗疲劳作用机制研究

目的 基于网络药理学方法探究刺五加总苷抗疲劳的潜在作用机制.方法 以刺五加总苷中7种主要活性成分为研究对象,利用Swiss Target Prediction网络平台预测其作用靶点;通过GeneCards、OMIM、DiGseE数据库获取与疲劳有关的靶标,借助Cytoscape 3.7.2软件构建"活性成分-抗疲劳靶点...     

三颗针炮制前后对小鼠抗疲劳抗氧化影响

目的:研究三颗针炮制前后提取物对小鼠抗疲劳和抗氧化的影响。方法:通过力竭游泳运动(ESE)评估三颗针炮制前后不同剂量(0.2、0.4、0.8 mg/g)提取物的抗疲劳作用;测定ESE后与疲劳相关的生化参数和活性提取物的体外抗氧化活性。结果:三颗针酒制品显著延长了小鼠游泳力竭的时间(P<0.05),表明酒制三颗针具有抗疲劳作用;酒制品组小鼠葡萄糖(Glu)、肌糖原水平显著升高,血尿素氮(BUN)、乳酸(Lac)水平显著降低。ESE增加了肌酸磷酸激酶(CK)、乳酸脱氢酶(LDH)、丙二醛(MDA)的水平,降低了超氧化物歧化酶(SOD)、谷胱甘肽(GSH)的水平,而酒制品可抑制上述变化。且体外抗氧化试验表明酒制品以剂量依赖性方式清除·OH和DPPH自由基。结论:三颗针酒制品可通过增加能量来源和减少有效代谢物的积累来缓解疲劳,抗氧化活性可能是酒制三颗针抗疲劳作用的原因之一。     

刺五加增强小鼠睡眠剥夺模型免疫功能和抗疲劳能力的实验研究

目的:探讨刺五加对小鼠睡眠剥夺模型免疫功能和抗疲劳能力的影响。方法:将小鼠随机分为正常对照组、模型组和刺五加低、高剂量组。刺五加低、高剂量组分别按照500,1 000 mg·kg-1的剂量ig,连续给药10 d,正常对照组和模型组每天ig等体积的蒸馏水。在第11 d,将模型组和刺五加低、高剂量组的小鼠采用单平台水环境法进行睡眠剥夺(sleepdeprivation,SD),在SD期间,继续给予实验药物。SD连续3 d后,采用炭粒廓清实验观察药物对SD小鼠免疫功能的影响,通过负重游泳实验和转棒疲劳实验观察药物对SD小鼠抗疲劳能力的影响。结果:炭粒廓清实验中,刺五加500,1 000 mg·kg-1的K值和α值为(0.021 4±0.008 3,0.030 7±0.005 9),(4.0±0.2,4.6±0.2),与SD组相比均显著增加;抗疲劳实验中,刺五加500,1 000 mg·kg-1的游泳时间和转棒时间为(828±83),(990±64)S,(330±63),(665±170)s,与SD组相比均显著延长。结论:刺五加具有增强睡眠剥夺小鼠免疫功能和抗疲劳能力的作用。     

抗疲劳中药对小鼠抗疲劳及调节免疫作用的研究

目的:探讨益气健脾、补肾调理复方中药对小鼠抗疲劳能力和免疫调节的作用。方法:6周龄清洁级昆明种小鼠随机分为4组:生理盐水组对照组、抗疲劳中药复方高、中、低剂量组,小鼠进行适应性运动训练1周,进行动物实验,每日灌胃1次,用药2周,通过爬竿试验、游泳试验、吞噬指数、免疫器官质量法(胸腺指数和脾脏指数)观察益气健脾、补肾调理中药复方抗疲劳的作用和增强免疫功能的作用。结果:抗疲劳中药复方能延长小鼠负重游泳的时间、延长爬竿时间,尤其是高剂量抗疲劳中药复方组能明显延长小鼠负重游泳持续时间,与生理盐水组比较有显著性差异,(P0.05);抗疲劳中药复方能增强巨噬细胞吞噬功能、脾脏指数(P0.05)。结论:抗疲劳中药复方具有增强小鼠抗疲劳能力和免疫功能作用。     

Acanthopanax senticosus extracts have a protective effect on Drosophila gut immunity

Ethnopharmacological relevance

Aanthopanax senticosus (A. senticosus) Harms is a classical adaptogenic agent used in China. It has been applied as an analeptic aid to improve weakened physical status. However, little is known about the effects of A. senticosus on inflammatory disease processes.

Materials and methods

Flies fed with standard cornmeal–yeast medium were used as controls, and the treatment groups contained 10% of A. senticosus aqueous extracts (root or fruit) in standard medium. Survival rate was performed by feeding a vial containing five layers of filter paper hydrated with 5% sucrose solution contaminated with pathogenic or toxic compounds. Imaging of the guts was viewed under the microscope. Death cells were detected by 7-AAD staining.

Results

The A. senticosus extract improved the survival rate, attenuated the death of intestinal epithelial cells, promoted the expression of antimicrobial peptide genes, and decreased the formation of melanotic masses. Moreover, our results indicated that the protective effect of fruit is much higher than that of root extracts.

Conclusions

A. senticosus extracts have a protective effect on Drosophila gut immunity and stress response, and may contribute to the prevention of inflammatory diseases induced by pathogenic and toxic compounds.     

Anticancer potential of Hericium erinaceus extracts against human gastrointestinal cancers

Ethnopharmacological relevance

Hericium is a genus of mushrooms (fungus) in the Hericiaceae family. Hericium erinaceus (HE) has been used for the treatment of digestive diseases for over 2000 years in China. HE possesses many beneficial functions such as anticancer, antiulcer, antiinflammation and antimicrobial effects, immunomodulation and other activities. The aim of the studies was to evaluate the anticancer efficacy of two extracts (HTJ5 and HTJ5A) from the culture broth of HE against three gastrointestinal cancers such as liver, colorectal and gastric cancers in both of in vitro of cancer cell lines and in vivo of tumor xenografts and discover the active compounds.

Materials and methods

Two HE extracts (HTJ5 and HTJ5A) were used for the studies. For the study of chemical constituents, the HTJ5 and HTJ5A were separated using a combination of macroporous resin with silica gel, HW-40 and LH-20 chromatography then purified by semipreparative high-performance liquid chromatography (HPLC) and determined by nuclear magnetic resonance (NMR) spectra. For the in vitro cytotoxicity studies, HepG2 and Huh-7 liver, HT-29 colon, and NCI-87 gastric cancer cell lines were used and MTT assay was performed to determine the in vitro cytotoxicity. For in vivo antitumor efficacy and toxicity studies, tumor xenograft models of SCID mice bearing liver cancer HepG2 and Huh-7, colon cancer HT-29 and gastric cancer NCI-87 subcutaneously were used and the mice were treated with the vehicle control, HTJ5 and HTJ5A orally (500 and 1000 mg/kg/day) and compared to 5-fluorouraci (5-FU) at the maximum tolerated dose (MTD, 25–30 mg/kg/day) intraperitoneally daily for 5 days when the tumors reached about 180–200 mg (mm³). Tumor volumes and body weight were measured daily during the first 10 days and 2–3 times a week thereafter to assess the tumor growth inhibition, tumor doubling time, partial and complete tumor response and toxicity.

Results

Twenty-two compounds were obtained from the fractions of HTJ5/HTJ5A including seven cycli dipeptides, five indole, pyrimidines, amino acids and derivative, three flavones, one anthraquinone, and six small aromatic compounds. HTJ5 and HTJ5A exhibited concentration-dependent cytotoxicity in vitro against liver cancer HepG2 and Huh-7, colon cancer HT-29, and gastric cancer NCI-87 cells with the IC₅₀ in 2.50±0.25 and 2.00±0.25, 0.80±0.08 and 1.50±0.28, 1.25±0.06 and 1.25±0.05, and 5.00±0.22 and 4.50±0.14 mg/ml; respectively. For in vivo tumor xenograft studies, HTJ5 and HTJ5A showed significantly antitumor efficacy against all four xenograft models of HepG2, Huh-7, HT-29 and NCI-87 without toxicity to the host. Furthermore, HTJ5 and HTJ5A are more effective than that of 5-FU against the four tumors with less toxicity.

Conclusion

HE extracts (HTJ5 and HTJ5A) are active against liver cancer HepG2 and Huh-7, colon cancer HT-29 and gastric cancer NCI-87 cells in vitro and tumor xenografts bearing in SCID mice in vivo. They are more effective and less toxic compared to 5-FU in all four in vivo tumor models. The compounds have the potential for development into anticancer agents for the treatment of gastrointestinal cancer used alone and/or in combination with clinical used chemotherapeutic drugs. However, further studies are required to find out the active chemical constituents and understand the mechanism of action associated with the super in vivo anticancer efficacy. In addition, future studies are needed to confirm our preliminary results of in vivo synergistic antitumor efficacy in animal models of tumor xenografts with the combination of HE extracts and clinical used anticancer drugs such as 5-FU, cisplatin and doxurubicin for the treatment of gastrointestinal cancers.     

基于UPLC-Q-TOF-MS及数据自动处理技术的 刺五加提取物中异嗪皮啶及其代谢产物分析

目的:分析大鼠灌胃刺五加提取物后血浆、胆汁、尿液和粪便中异嗪皮啶(M0)及其代谢产物。方法:健康雄性wistar大鼠按325 mg.kg-1的剂量灌胃给予刺五加提取物,分别采集给药后1,1.5,2,4,6 h肝门静脉血液并用SPE技术制备血浆;采集0～12 h胆汁并用SPE技术制备;0～12 h,12～24 h分别采集尿液和粪便样品并制备样品,采用UPLC-Q-TOF-MS技术和Metabolynx XS软件联合的方法分析。结果:在大鼠血浆、胆汁、尿液和粪便中检测到M0,在血浆和胆汁中检测到代谢物异嗪皮啶葡萄糖醛酸苷(M1)。其中M1是首次报道的异嗪皮啶代谢产物。结论:异嗪皮啶在大鼠体内以葡萄糖醛酸形式代谢,最终又以异嗪皮啶形式排出体外。     

Fundamental studies on the inhibitory action of Acanthopanax senticosus Harms on glucose absorption

Aim of the study

Acanthopanax senticosus Harms extract (ASE) is used as an ingredient of over-the-counter drugs and functional foods, such as health supplements, in Japan. ASE exhibits a hypoglycemic effect; however, the mechanism of the hypoglycemic effect is not clear. In the present study, we investigated whether ASE has a glucose absorption inhibitory action.

Materials and methods

We examined the effects of ASE on α-amylase and α-glucosidase activities, and on glucose uptake in Caco-2 cells. We also examined the effects of ASE oral administration on glucose tolerance in type 2 diabetes mellitus model db/db mice.

Results

The addition of ASE inhibited α-glucosidase activity but not α-amylase activity. The α-glucosidase inhibitory activity of ASE was approximately 1/13 of that of acarbose. The addition of ASE inhibited 2′-deoxy-d-glucose (DG) uptake in human intestinal Caco-2 cells, and the inhibitory activity of ASE was approximately 1/40 of that of phloretin. Kinetic analysis of glucose uptake indicated that ASE has no effects on DG uptake through passive diffusion, but that ASE inhibits intracellular DG uptake chiefly by inhibiting transport via a glucose transporter. In the glucose tolerance study, db/db mice orally administered ASE for 3 days showed significantly lower plasma glucose level than the control group 30 min after sucrose loading, without affecting plasma insulin levels. In addition, ASE oral administration significantly inhibited α-glucosidase activity in the small intestine mucosa extirpated from the mice.

Conclusion

These findings indicate that ASE may be useful as an ingredient of functional foods to improve postprandial hyperglycemia and prevent type II diabetes mellitus.     

刺五加胶囊成型工艺

目的:探讨刺五加胶囊的成型工艺。方法:考察不同辅料对主药吸湿性的影响;以颗粒的合格率作为指标,筛选最佳制粒成型工艺条件,并考察颗粒的临界相对湿度,休止角及堆密度。结果:主药与可溶性淀粉、微晶纤维素按40∶45∶15的比例混匀,加入70%乙醇制软材,20目筛制粒,所得颗粒的休止角为33.5°,堆密度为0.38 g.mL-1,临界相对湿度为66%。结论:该成型工艺合理、可行,为大生产提供了科学依据。     

Protective effects of aqueous extract from Acanthopanax senticosus against corticosterone-induced neurotoxicity in PC12 cells

Ethnopharmacological relevance

Acanthopanax senticosus, classified into the family of Araliaceae, has been known for thousands of years as a remedy and is used to treat various diseases in traditional Chinese medicine system including hypertension, ischemic heart disease and hepatitis.

Aim of the study

This study aimed to examine the protective effects of aqueous extract from Acanthopanax senticosus (ASE) on corticosterone-induced neurotoxicity and its possible mechanisms, using PC12 cells as a suitable in vitro model of depression.

Materials and methods

In this paper, PC12 cells were treated with 200 μM of corticosterone in the absence or presence of ASE in varying concentrations for 24 h. Then, cell viability was measured by MTT assay. The release amount of lactate dehydrogenase (LDH) was quantified using LDH assay kit. Apoptosis of PC12 cells was measured by Annexin V-FITC and PI labeling. The intracellular Ca²⁺ content was tested by fluorescent labeling. The mRNA level of brain-derived neurotrophic factor (BDNF) was examined by real-time RT-PCR, and the expression of cAMP response element binding protein (CREB) was determined by western blotting.

Results

The results showed that treatment with 200 μM of corticosterone could induce cytotoxicity in PC12 cells. However, different concentrations of ASE (50, 100, 200, and 400 μg/mL) significantly increased the cell viability, decreased the LDH release, suppressed the apoptosis of PC12 cells, attenuated the intracellular Ca²⁺ overloading, up-regulated the BDNF mRNA level and CREB protein expression compared with the corresponding corticosterone-treated group.

Conclusion

The present results suggest that ASE exerts a neuroprotective effect on corticosterone-induced neurotoxicity in PC12 cells, which may be one of the acting mechanisms that accounts for the in vivo antidepressant activity of ASE.     

猴头菌粉提取物对2型糖尿病小鼠降血糖作用研究

目的:研究猴头菌粉提取物的降血糖活性.方法:健康雄性昆明小鼠,以4周高脂饮食诱发胰岛素抵抗,并以ip链脲佐菌素(STZ) 170 mg?kg-1以诱发2型糖尿病.模型小鼠随机分组,ig给予猴头菌粉各部位提取物21 d,测定空腹血糖以筛选猴头菌粉降血糖活性部位,对筛选出的活性部位进行口服糖耐量实验.并于HE染色后,在光镜下观察其对糖尿病模型小鼠肝、肾、胰腺的组织形态学影响.结果:猴头菌粉多糖能对抗糖尿病小鼠体重减轻的症状,给予猴头菌粉多糖的14,21 d小鼠血糖值与模型组相比较均出现非常显著性下降(P＜0.01).猴头菌粉多糖(400 mg?kg-1)能提高糖尿病模型小鼠的葡萄糖耐受量,并能对抗糖尿病导致的肝、肾、胰脏病变.结论:猴头菌粉多糖具有降血糖活性,且其提高糖耐量、保护脏器作用呈现一定的量效关系.     

川西高原红毛五加群落生态学研究

目的:研究川西高原地区红毛五加群落特征,红毛五加在群落中的地位与适应性。方法:采用群落生态学方法调查不同红毛五加样地植被组成、红毛五加丛数与无性分株数,计算各物种重要值、各样地生物多样性指数,统计不同群落生活型谱。采用卡方检验分析红毛五加对群落内主要灌木物种的依存度。结果:红毛五加生境的群落类型可分为3类,不同生境类型中红毛五加种群数目差异较大,红毛五加与群落中主要灌木物种不呈显著性种间关联。结论:红毛五加种群生长对光照因子敏感,红毛五加可能具备单一种群栽培的生物学特性。