Reveal the pharmacodynamic substances and mechanism of an edible medicinal plant Rhodiola crenulate in DSS-induced colitis through plasma pharmacochemistry and metabolomics

Rhodiola crenulate is the edible medicinal herbal medicine widely used for altitude sickness in China.Interestingly,our previous work has found that R.crenulate extract(RCE)could significantly improve the pathology associated with dextran sulfate sodium-induced colitis.Thus,the current research aims to reveal the pharmacodynamic material basis of RCE,as well as its mechanism against colitis.The chemical characterization of RCE was performed by UHPLC-HR-MS,through which a total of 88 constituents were identified.Meanwhile,our results also found 29 constituents absorbed into blood and 8 metabolized absorbable compounds.The decreased flavonoids prototype and the elevated sulfated products of phenols were observed under pathophysiological conditions of colitis.The metabolomics study revealed that colitis caused the alternation of fatty acid metabolism,steroid hormone biosynthesis and bile acid metabolism.Correspondingly,RCE could prevent colitis by improving fatty acid metabolism and secondary bile acid metabolism.

A heteropolysaccharide from Rhodiola rosea L.:preparation,purification and anti-tumor activities in H22-bearing mice

Numerous polysaccharides isolated from plants have been used to augment traditional drugs in the treatment of cancer.In order to explore the influence to hepatocellular carcinoma,a novel cold water-soluble polysaccharide was separated from Rhodiola rosea L.root(RLP)and then its structure and anti-cancer activities were tested.The chemical compositions and high performance gel permeation chromatography(HPGPC)results indicated that RLP was an acid heteropolysaccharide with the molecular weight of about1.15×10~6 Da.Furthermore,ion chromatography(IC),Fourier transform infrared(FT-IR)and nuclear magnetic resoance(NMR)further indicated that RLP was main composed of→2,4)-α-Rha(1→,→5)-α-L-Araf-(1→,α-D-Glu,→6)-β-D-Galp-(1→,β-D-Man and→4)-α-GalpA-(1→.In vivo antitumor activities of RLP were carried out by using H22 tumor-bearing mice model.The results shown that RLP(100 and 300 mg/kg)could inhibit tumor growth of H22 cells from 23.59%to 45.52%and protect thymuses and spleen without damage.In addition,according to cell cycle,AV-FITC/PI and JC-1,RLP could induce dose-dependent apopto sis of H22 cells via S phase arrested which was through a mitochondrial related pathway.Our data indicated that RLP has a broader application prospect in anti-tumor preparations.

红景天(Rhodiola L)的研究进展

文章综述了红景天研究概况,主要介绍了红景天的主要化学成分及药用价值,其中药用价值主要包括抗衰老作用、抗缺氧作用、双向调节作用、抗疲劳作用、抗辐射作用及抗肿瘤作用,并就红景天开发中存在的问题及未来研究的方向提出建议。

长鞭红景天(Rhodiola fastigiata)内生细菌的分离及其生理生化特性测定

从西藏自治区林芝地区色季拉山采集长鞭红景天(Rhodiola fastigiata)植株,从中共分离到内生细菌15株,并根据细菌菌落特征,将15株细菌归为9种。另外,对9种红景天内生细菌的革兰氏染色反应、接触酶活性、淀粉水解、纤维素水解、明胶液化、牛奶分解、甲基红反应、抗生素敏感性等生理生化特性进行了测定,为长鞭红景天(Rhodiola fastigiat)内生菌的开发利用奠定了基础。

Preliminary Study on the ITS Sequences of nrDNA from Rhodiola alsia

[Objective] The study aimed to analyze the ITS sequences of nrDNA from Rhodiola alisa and investigate the difference of evolution rate between nrDNA and trnS-trnG and rpl20-rps12 sequences of cpDNA(chloroplast DNA).[Method]Total DNA was extracted from silica-dried leaves of R.alsia by using modified CTAB method.With the extracted DNA sample as template,nrDNA ITS region was amplified,then purified and sequenced.In addition,the yielded ITS sequences were also compared with the known trnS-trnG and rpl20-rps12 sequences of cpDNA from R.alsia.[Result]The ITS sequence of nrDNA from R.alsia was 701 bp in length,of which 13 variable sites were found with a percentage of 1.85%.Of the 13 variable sites,8 were caused by point mutations,5 were the results of insertions or deletions.The(A+T)content and(G+C)content were 46.9% and 53.1%,respectively.The nucleotide diversity(π)was 0.004 27.[Conclusion]The ITS region of nrDNA from R.alsia was more conservative and evolved more slowly than the trnS-trnG and rpl20-rps12 sequences of its cpDNA.

A New Glucoside from Rhodiola fastigiata (Crassulaceae)

A new glucoside, fastigitin A (1), namely 2-O-β-D-glucopyransyl-3-methyl-methyl pinalate, together with twelve known constituents (2-13), was isolated from the root of Rhodiola fastigiata (Hook. f. et Thoms.) S. H. Fu collected from Nujiang Lisu autonomous region, Yunnan, China. Their structures were identified by spectral (including 2D-NMR techniques) and chemical methods. Compounds 2 and 5-9 were obtained from this plant for the first time.

高山红景天(Rhodiola sachalinensis)试管苗缓慢生长法保存及试管苗DNA含量分析

为建立高山红景天试管苗的缓慢生长法保存体系,在低温、弱光及不同浓度甘露醇条件下对两种来源的试管苗进行缓慢生长法保存,并对其恢复生长植株的DNA含量进行分析.结果表明:低温(6±1)℃和弱光600 lx(8 h·d-1)条件下,试管苗在添加有甘露醇10 g·L-1的1/2MS+蔗糖20 g·L-1+琼脂7 g·L-1培养基中可保存18个月,平均存活率在85%以上,且恢复生长后试管苗长势良好.上述方法保存后恢复生长的植株经流式细胞仪检测表明核DNA含量没有发生改变.该结果为高山红景天种质资源缓慢生长法保存的有效性和可行性提供了理论依据.

高山红景天(Rhodiola sachalinensis A.Bor)茎尖包埋-玻璃化法超低温保存与植株再生

使用包埋-玻璃化法对两种来源的高山红景天茎尖进行超低温保存研究，探讨蔗糖预处理、包埋过程中渗透处理和不同温度下玻璃化液PVS2处理的时间对超低温保存后茎尖存活率的影响．结果表明，茎尖依次在0．3、0．5、0．7、1．0mol·L^-1的蔗糖溶液中各预培养1d后，转入1．0mol·L^-1的蔗糖溶液中继续培养4d，然后使用附加2mol·L^-1甘油和1mol·L^-1蔗糖的包埋液渗透处理60min，包埋珠在0℃处理180～210min后投入液氮，48h后用40℃水浴快速化冻3min，用含有1mol·L。蔗糖的改良MS培养液洗涤20min，转入MS＋6-BA1mg·L^-1＋NAAO．1mg·L^-1固体培养基中在22℃条件下进行暗培养，2d后转移入光照培养，最高成活率接近100％，再生植株生长和分化正常．

高山红景天(Rhodiola sachalinensis)遗传变异及其濒危机制的探讨

采用淀粉凝胶电泳手段，等位酶分析技术对分布于黑龙江大海林林业局高山红景天天然分布区内３个亚种群及吉林长白山１个亚种群的９种酶系统１８个位点进行了遗传分析，结果表明种群内存在一定的遗传变异（多态位点百分比为３０．５５％，等位基因平均数为１．３３３），但不是很丰富，主要是该种狭窄的分布区致使生境单一及人为破坏其本身的生物学特性在长期的进化过程中相互适应相互选择的结果。在黑龙江的分布区内各亚种群之间一致度较高，而位于长白山的亚种群与黑龙江的各亚种群之间有一定的遗传分化，而且个别位点上差异很大，这可能与适应特定的环境有关。

Pretreatment with Rhodiola Rosea Extract Reduces Cognitive Impairment Induced by Intracerebroventricular Streptozotocin in Rats: Implication of Anti-oxidative and Neuroprotective Effects

Objective To investigate the pretreatment effects of Rhodiola rosea （R. rosea） extract on cognitive dysfunction, oxidative stress in hippocampus and hippocampal neuron injury in a rat model of Alzheimer＇s disease （AD）. Methods Male Sprague-Dawley rats were pretreated with R. rosea extract at doses of 1.5, 3.0, and 6.0 g/kg for 3 weeks, followed by bilateral intracerebroventricular injection with streptozotocin （1.5 mg/kg） on days 1 and 3. Behavioral alterations were monitored after 2 weeks from the lesion using Morris water maze task. Three weeks after the lesion, the rats were sacrificed for measuring the malondialdehyde （MDA）, glutathione reductase （GR） and reduced glutathione （GSH） levels in hippocampus and histopathology of hippocampal neurons. Results The MDA level was significantly increased while the GR and GSH levels were significantly decreased with striking impairments in spatial learning and memory and severe damage to hippocampal neurons in the model rat induced by intracerebroventricular injection of streptozotocin. These abnormalities were significantly improved by pretreatment with R. rosea extract （3.0 g/kg）. Conclusion R. rosea extract can protect rats against cognitive deficits, neuronal injury and oxidative stress induced by intracerebroventricular injection of streptozotocin, and may be used as a potential agent in treatment of neurodegenerative diseases such as AD.

Free radical scavenging property and antiproliferative activity of Rhodiola imbricata Edgew extracts in HT-29 human colon cancer cells

Objective:To investigate the in vitro antioxidant and antiproliferative activity of rhizome extracts of Rhodiola imbricata(R.imbricata) in HT-29 human colon cancer cell line.Methods:The successively extracted rhizome of R.imbricata using various solvents was analyzed for their total phenolics,tannins and flavonoid contents.In vitro antioxidant activity was evaluated by employing different assays,including DPPH,ABTS radical scavenging assays,FRAP, phosphomolybdenum reduction assay,superoxide anion,hydroxyl radical scavenging activities and metal chelating ability.Results:Acetone and methanol extracts recorded higher phenolic content and showed comparable antioxidant activity with standard reference.Additionally,they also inhibited the proliferation of HT-29 cells upon treatment at higher concentration(200μg/mL) (acetone and methanol,84%and 84%.respectively).On examination acetone extract exhibited antiproliferative activity in a concentration dependent manner whereas,methanol extract showed both dose dependent and lime dependent inhibitory activity.Conclusions:The results obtained justify the traditional usage of R.imbricata from their promising antioxidant activity.

Gas exchange of Rhodiola sachalinensis transplanted from different habitats in Changbai Mountain

Rhodiola sachalinensis growing in the different habitats, Xiaotianchi plot (altitude 1800 m), Tree line plot (altitude 2000 m) and Tianwenfeng plot (altitude 2325 m), of Changbai Moutain (northern slop) were transplanted to Harbin Experimental Forest Farm for measuring its characteristic of gas exchange. The study results indicated that the growth state and gas exchange characteristics ofRh. sachalinensis growing in different habitats varied markedly. The plants transplanted from Tianwenfeng plot had the highest values in net photosynthesis rate (Pn) and transpiration rate (Tr), those transplanted from Tree line plot shows the second, and those transplanted rom Xiaotianchi plot had the lowest values. The variance existed in transplanted plants was the same as shown in the field. From the result it can be extrapolated that the difference of Pn and Tr existed inRh. sachalinensis transplanted from different habitats is depend not only on the environmental factors, but on the variance of physiological characteristic of plant itself.

Diurnal change of gas exchange of Rhodiola sachalinensis transplanted from different habitats in Changbai Mountain

Rhodiola sachalinensis growing in the different habitats, such as Xiaotianchi plot (altitude 1800 m), Tree line plot (altitude 2000 m) and Tianwenfeng plot (altitude 2325 m) of Changbai Mountain (northern slop) were transplanted to Harbin Experimental Farm for determining its gas exchange. The study results indicated that the diurnal changes of gas exchange ofRh. sachalinensis transplanted from different habitats were still different though the morphological appearance of the newly sprouted above-ground part had become more similar. For net photosynthesis rate (Pn), stomatal conductance (Gs) and apparent quantum yield (AQY), the diumal change curves ofRh. sachalinensis transplanted from the three plots were similar, but the data value were different. For the dark respiration rate (Rd), transpiration rate (Tr) and water use efficiency (WUE), diurnal change curves of the three plots were obviously different.

Identification of Rhodiola species by using RP-HPLC

An approach was established using RP-HPLC (reversed-phase high-performance liquid chromatography) to identify ten species of Rhodiola, R. coccinea A. Bor, R. junggarica C.Y. Yang et N.R. Cui spn., R. heterodonta A. Bor, R. linearifolia A. Bor, R. pamiro alaiucm A. Bor, R. kaschgarica A. Bor, R. litwinowii A. Bor, R. gelida schrenk, R. rosea L. and R. quadrifide Fisch et Mey collected from the Tianshan Mountains areas of China. Chromatograms of alcohol-soluble proteins, generated from these ten Rhodiola spp. were compared. Each chromatogram of alcohol-soluble proteins came from a single seed of one wild species only. The results showed that when using a Waters Delta Pak. C18, 5 μm particle size reversed phase column (150 mm×3.9 mm), a linear gradient of 22%?55% solvent B with a flow rate of 1 ml/min and a run time of 67 min, the chromatography gave optimum separation of Rhodiola alcohol-soluble proteins. Chromatogram of each species was different and could be used to identify those species. Cluster analysis of genetic similarity coefficients of 37% to 60% showed a medium degree of genetic diversity among the species in these eco-areas. Cluster analysis showed that the ten species of Rhodiola can be divided into four clusters and yielded the general and unique biochemical markers of these species. RP-HPLC was shown to be a rapid, repeatable and reliable method for Rhodiola species identification and analysis of genetic diversity.

Effect of Ethanol Extract of Rhodiola Rosea on the Early Nephropathy in Type 2 Diabetic Rats

Summary： This study aimed to investigate the therapeutical effects of Rhodiola rosea extract on rats with type 2 diabetic nephropathy （DN）. The rat type 2 DN model was established by high fat and high calorie feeding and intravenous injection of streptozocin （STZ）. Wistar rats were randomly divided into normal group, control group, low dose Rhodiola rosea group, high dose Rhodiola rosea group and Cap- topril group. Oral glucose tolerance test （OGTT） was performed to determine the impairment of glucose tolerance in the established animal model. A series of parameters including fasting blood glucose （FBG）, total cholesterol （TC）, triglyceride （TG）, creatinine clearance rate （Ccr）, 24-h urinary albumin （UA）, the ratio of kidney mass/body weight （renal index） and glomerular area were examined after 8 weeks. Moreover, the expression of transforming growth factor （TGF）-β1 in renal tissues was detected by using immunohistochemisty. At the end of the eighth week, FBG, TC, TG, Ccr, 24-h urinary albumin, the ratio of kidney mass/body weight and glomerular area were significantly reduced in Rhodiola rosea extract treatment groups as compared with those in control group. TGF-β1 expression in renal tissues of Rhodiola rosea extract treatment groups was also significantly decreased as compared with that of con- trol group. These results indicate that Rhodiola rosea extract may have a protective effect on early nephropathy in diabetic rats, which might be related to the decrease of the renal expression of TGF-β1.

Discovery of human pancreatic lipase inhibitors from root of Rhodiola crenulata via integrating bioactivity-guided fractionation,chemical profiling and biochemical assay

Although herbal medicines(HMs)are widely used in the prevention and treatment of obesity and obesity-associated disorders,the key constituents exhibiting anti-obesity activity and their molecular mechanisms are poorly understood.Recently,we assessed the inhibitory potentials of several HMs against human pancreatic lipase(hPL,a key therapeutic target for human obesity),among which the root-extract of Rhodiola crenulata(ERC)showed the most potent anti-hPL activity.In this study,we adopted an integrated strategy,involving bioactivity-guided fractionation techniques,chemical profiling,and biochemical assays,to identify the key anti-hPL constituents in ERC.Nine ERC fractions(retention time=12.5e35 min),obtained using reverse-phase liquid chromatography,showed strong anti-hPL activity,while the major constituents in these bioactive fractions were subsequently identified using liquid chromatography-quadrupole time-of-flight mass spectrometry(LC-Q-TOF-MS/MS).Among the identified ERC constituents,1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose(PGG)and catechin gallate(CG)showed the most potent anti-hPL activity,with pIC50 values of 7.59±0.03 and 7.68±0.23,respectively.Further investigations revealed that PGG and CG potently inhibited hPL in a non-competitive manner,with inhibition constant(Ki)values of 0.012 and 0.082 mM,respectively.Collectively,our integrative analyses enabled us to efficiently identify and characterize the key anti-obesity constituents in ERC,as well as to elucidate their anti-hPL mechanisms.These findings provide convincing evidence in support of the anti-obesity and lipid-lowering properties of ERC.

The neuroprotective effect of Rhodiolae on dementia rat induced by ibotenic acid

OBJECTIVE To research the effect of Rhodiolae on cognitive function on dementia rats and to investigate its potential mechanism.METHODS Established dementia model via injecting ibotenic acid into basal nuclei,and randomly divided the model rats into model group,Aricept group(1.4 mg·kg-1)and Rhodiolae group(0.14 mg·kg-1),moreover the sham group was built byinjecting normal saline.All animals received homologous drugs after modeling 1 week,and the sham group and model group were gave isometric normal saline(10 mL·kg-1),1/d.Morris water maze was used to evaluate the change of cognitive function after the rats were treated 40 d,and pathomorphism in CA1 and cerebral cortex were observed by Nissl staining and the numbers of nerve cell in these areas were counted.The concentration of MDA,H2O2,CAT,SOD and GR in brain tissue were detected by spectrophotography.RESULTS Compared to model group,the escape latency of Rhodiolae groups〔(31.10±20.97)s compare to(51.07±13.04)s〕was shortened but it hadn′t statistical difference,and the total swimming distance〔(8469.88±4724.74)cm compare to(15862.17±1315.69)cm〕was shortened obviously,furthermore the times entered the target quadrant〔(9.50±1.87)times compare to(6.50±1.38)times〕and the percentage in target quadrant〔(33.87±4.47)%compare to(26.17±2.58)%〕were evidently increased.Moderate shrinkage of nucleus of neurons in cerebral cortex of model rats,mild hyperplasia of glial cells and loosely neuron was also observed in CA1 area,but those lesions were distinctly ameliorated in Rhodiolae groups.What′s more,the numbers of nerve cells in CA1〔(70.40±4.00)compare to(45.50±4.95)〕and cerebral cortex〔(63.00±5.29)compare to(42.40±11.87)〕of Rhodiolae groups were obviously increased.In addition,compared to model group,the content of MDA〔(14.72±4.42)compare to(19.99±4.70)g protein·L-1〕and H2O2〔(60.07±6.38)mmol·L-1compare to(76.37±6.06)mmol·L-1〕were evidently decreased,the activity of GSH〔(41.30±14.81)k U·g-1protein compare to(15.93±3.59)k U·g-1protein〕was visibly increased,and the activity of SOD〔(5.74±3.76)g protein·L-1compare to(2.72±1.09)g protein·L-1〕and CAT〔(210.45±60.11)k U·g-1protein compare to(171.48±56.50)k U·g-1protein〕were also increased but without statistical difference.CONCLUSION Rhodiolae improve learning and remembering function,ameliorate pathological changes and protect neuronal loss which respond to decrease the oxidative products and enhance the activity of antioxidase.

Analysis on Chemical Constituents of Rhodiola crenulata

[Objectives] To measure and analyze chemical composition of R. crenulata. [Methods] Taking R. crenulata in Lang County of Tibet as the research object,the routine nutritional components,trace elements and amino acid were determined,and the nutrition was evaluated by the international general evaluation method. [Results]The protein content of R. crenulata in Lang County of Tibet was 2. 08%,and fat was 5. 67%. Amino acid was incomplete amino acid,and Ile,Met + Cys were the first limiting amino acids of rhodiola root protein,while Leu was the second limiting amino acid of rhodiola root protein. The final amino acid score was 0. Magnesium,iron,zinc and other mineral elements were abundant,and there was no mercury,cadmium,lead and other heavy metals residues. [Conclusions]The research could provide a powerful basis for industrialization development of rhodiola.

The Optimization of Ultrasonic Extraction Process of Total Flavonoids from Rhodiola Crenulata by Response Surface Method

[Objectives]To study the optimal extraction process of total flavonoids in Rhodiola rosea L.,and further facilitate the development and utilization of Rhodiola rosea L. [Methods]With Rhodiola crenulata as raw material,ethanol as extractant,ultrasonic extraction as the extraction method,the single factor method was first used for preliminary investigation of effect of ethanol volume fraction,solid-liquid ratio,extraction temperature and extraction time on the flavonoids extraction rate,and then the Box-Behnken response surface method was used to optimize the extraction process of total flavonoids in R. crenulata. [Results] The optimal extraction conditions: ethanol concentration of 72%;solid-liquid ratio of 1∶ 43; extraction temperature of 66℃; extraction time of 50 min. Under these conditions,the extraction rate of total flavonoids from R. crenulata was 2. 591%. [Conclusions] The results showed that the method of response surface was reasonable and feasible for the optimization of ultrasonic extraction of total flavonoids from R. crenulata.

Neuropharmacological Characterization of Extracts from Rhodiola rosea, Oenothera paradoxa and Paullinia cupana in Comparison to Caffeine

To find possible therapeutic applications involving the Central Nervous System (CNS) for herbals is a major challenge during functional food and drug discovery and development programmes. Despite the availability of numerous in vitro and in vivo tests, there is no single agreed screening procedure for pharmacological testing of herbal extracts with anticipated CNS activity. Experience gained from more than 25 years of testing has shown that two models give reasonably reliable orientation for future CNS applications: construction of an electropharmacogram based on wireless recording of field potentials from the depth of the brain of freely moving rats (Tele-Stereo-EEG) and recording of the population spike produced by pyramidal cells from hippocampal slices in vitro. A combination of these two methods has now been used to characterize the pharmacological profile of extracts from Rhodiola rosea root, Oenothera paradoxa seeds and Paullinia cupana seeds. Spectral analysis of field potentials revealed attenuation of alpha2 and beta1 waves was common for all extracts. According to previous studies, this is interpreted as activation of the dopaminergic and glutamatergic transmission. In addition, Oenothera and Rhodiola extracts attenuated delta and theta power, probably related to interference with the cholinergic and norepinephrinergic transmission, respectively. Using discriminant analysis for comparison with reference pharmaceutical and botanical drugs, Rhodiola projected near the position of Ginkgo extract, whereas Oenothera extract was projected near the position of Tramadol, an analgesic drug. Physical motion was increased only in the presence of Paullinia extract and caffeine. Increases of long-term potentiation were observed in the presence of Rhodiola extract, Paullinia extract and caffeine. The combined information predicts stimulant and cognitive function-enhancing activities in humans for the Rhodiola extract, which could also be used as a possible caffeine-replacement, and antidepressant and analgesic activity for the Oenothera extract.