Levisticum officinale extract protects against CCl4-induced hepatotoxicity through anti-inflammatory,anti-fibrotic,and antioxidant properties in rats

Objective:To investigate the hepatoprotective effects of Levisticum officinale extract on CCl4-induced hepatotoxicity.Methods:Different doses of Levisticum officinale extract were given orally to rats for 10 days,then rats received a single dose of CCl4(2.5 mL/kg,50%v/v in liquid paraffin).Biochemical and histopathological assays were performed to assess the effects of the extract on liver function and architecture.Moreover,antioxidant and oxidative markers as well as inflammatory and fibrotic indicators were measured.Results:Pretreatment with Levisticum officinale extract significantly mitigated CCl4-induced damage to liver structure,improved serum levels of alanine aminotransferase,aspartate aminotransferase,alkaline phosphatase,urea,total bilirubin,and total protein,enhanced glutathione content and superoxide dismutase and catalase activities in the liver,as well as decreased plasma and hepatic malondialdehyde levels.Immunohistochemical results demonstrated that the extract reduced Ki-67 andα-SMA expression and Masson’s trichrome staining revealed decreased liver collagen in rats treated with Levisticum officinale extract.Moreover,Levisticum officinale extract markedly decreased the gene expressions of TNF-α,IL-6,TGF-β1,MCP-1,and COX-2.Conclusions:Levisticum officinale extract exerts hepatoprotective effects on CCl4-induced hepatotoxicity through antioxidant,anti-inflammatory,and anti-fibrotic activities.

Implementation of the Acclimatization Process for Ginger Plantlets (Zingiber officinale Roscoe)

Ginger (Zingiber officinale Roscoe) is an important culinary and medicinal spice but is rarely cultivated due to the unavailability of seeds. Given the difficulties in adapting to plantlets produced in natural environments, it is important to analyze the survival conditions of ginger plantlets. For this reason, we varied the incubation temperature and humidity as well as the substrate during the weaning phase. Then, we varied the nutrients contained in the watering solution during the hardening phase. The statistical analysis showed that physical factors and substrates significantly influenced (p < 0.0001) plantlet survival. Nutrient solutions significantly influenced the phylogenesis, rhizogenesis, and height growth of the plantlets. The suitable physical factors for good development of plantlets are a temperature of 26.54˚C and a humidity of 96.16%. The 1C2T2TC substrate (1 Compost + 2 Soil + 2 Coconut Peat) had a significant survival rate of approximately 92.5%. During hardening, the Plantzym solution promoted good growth in terms of plantlet height (0.6 cm) and good development of roots (30 roots) and leaves (03 leaves). This work will make it possible to develop a technical seed production sheet for better development of ginger cultivation in Benin.

Functional analysis of flavanone 3-hydroxylase(F3H)from Dendrobium officinale,which confers abiotic stress tolerance

Flavonoids are important bioactive components in Dendrobium officinale,a medicinal orchid.They are involved in many biological activities,including protecting plants against biotic and abiotic stresses.Research on the key genes related to flavonoid biosynthesis in D.officinale is limited.In this study,one of the key flavonoid biosynthesis genes,flavanone 3-hydroxylase(F3H),was characterized from D.officinale.The open reading frame of DoF3H was 1134 bp long and it encoded a 377-amino acid protein.The DoF3H protein showed considerably high homology with F3H proteins from other plant species and shared a common evolutionary ancestor with other F3Hs.DoF3H transcripts were detected in different organs of adult plants and mainly accumulated in flowers,followed by roots,stems and leaves,a pattern that was similar to the content of flavonoids.Recombinant DoF3H protein,which was localized in the cytosol,could convert naringenin to dihydrokaempferol.The mRNA levels of DoF3H were significantly induced by salt and cold stresses.Furthermore,the heterologous expression of DoF3H in Escherichia coli conferred it higher tolerance to salt and cold stresses.These results provide insight into the molecular function of DoF3H in the biosynthesis of flavonoids,and provide a new application for improvement of abiotic tolerance in D.officinale.

Study on Mechanism of Dendrobium officinale Kimura et Migo in Preventing and Treating Exercise-induced Muscle Damage(EIMD)in Rats

[Objectives]This study was conducted to observe the mechanism of Dendrobium officinale Kimura et Migo on gastrocnemius muscle in rats with exercise-induced muscle damage(EIMD).[Methods]The micro-injury model of skeletal muscle was established by treadmill training.Forty two SD rats were randomly divided into a control group,1,12 and 24 h exercise groups,D.officinale 2 ml+1 h exercise group,D.officinale 2 ml+12 h exercise group,and D.officinale 2 ml+24 h exercise group,with 6 rats in each group.Various D.officinale groups were given the drug once in the morning and once in the evening at a dose of 2 ml/time,a week in advance.Except for the quiet group,the samples were collected from the 1,12 and 24 h exercise groups after anesthesia following 1,12 and 24 h of exercise for the last time,respectively,and the D.officinale 2 ml+1 h exercise group,D.officinale 2 ml+12 h exercise group and D.officinale 2 ml+24 h exercise group were also sampled after anesthesia following 1,12 and 24 h of exercise for the last time,respectively.The contents of ATP,CK-MM and CK in rat serum were determined by enzyme-linked immunosorbent assay(ELISA).The histopathological changes of gastrocnemius muscle were observed by HE staining.PCR and Western-blot detection were carried out to analyze the effects of D.officinale on IGF-1 mRNA and protein expression in gastrocnemius muscle.[Results]Compared with the quiet group,the ATP contents in the serum of rats in the 1,12 and 24 h exercise groups significantly decreased(P<0.01),while the CK and CK-MM contents significantly increased(P<0.01).The expression of IGF-1 mRNA and protein in the gastrocnemius muscle tissue significantly increased(P<0.01).Compared with the 1 h exercise group,the ATP content and IGF-1 protein expression in the gastrocnemius muscle tissue of the D.officinale liquid+1 h exercise group significantly increased(P<0.05),while the CK and CK-MM contents significantly decreased(P<0.01).Compared with the 12 h exercise group,the D.officinale liquid+12 h exercise group showed a significant increase in ATP content(P<0.01),significant increases in IGF-1 mRNA and protein expression in the gastrocnemius muscle tissue(P<0.01),and significant decreases in CK and CK-MM contents(P<0.01).Compared with the 24 h exercise group,the ATP content and IGF-1 mRNA and protein expression in the gastrocnemius muscle tissue of the D.officinale liquid+24 h exercise group significantly increased(P<0.01),while the CK and CK-MM contents significantly decreased(P<0.01).From the pathological tissue morphology of the gastrocnemius muscle in rats with EIMD treated with D.officinale,it could be concluded that the gastrocnemius muscle of each exercise group was significantly damaged,and the damage was significantly alleviated after administration of D.officinale liquid.[Conclusions]The effects and mechanism of D.officinale on prevention and treatment of EIMD in rats might be related to the promotion of IGF-1 mRNA and protein expression in injured tissues by reducing ATP energy consumption,CK-MM and CK activity.

Inoculation Effects of Dendrobium officinale Mycorrhizal Fungi on Their Plantlets

[Objective] This study aimed to explore the inoculation effects of Dendrobium officinale mycorrhizal fungi on their plantlets. [Method] Endophytic strains Tj1, Tj2 and Tj3 were obtained by isolation and purification from mycorrhiza of wild Dendrobium officinale and inoculated on the root system of Dendrobium officinale for inoculation test. [Result] Under tissue-culture conditions, at early stage, Tj1 strain hadn＇t shown promotion effect on Dendrobium officinale, Tj2 strain had shown relatively strong promotion effects, and Tj3 strain had promoted the growth of roots; at late stage, Tj1 strain had shown relatively strong promotion effects, Tj2 strain had shown the best inoculation effects and the strongest promotion effects, while Tj3 strain had caused root and seedling rot problems of the plantlets; under outdoor conditions, after inoculation with Tj2 strain, the number of leaves and lateral buds were increased, the growth of lateral root and the increase of plant height were significant, the leaves of Dendrobium officinale plantlets were large and dark green and an obvious root enlargement phenomenon was observed. [Conclusion] The two inoculation methods both indicate that Tj2 strain has relatively strong promotion effects on the growth of Dendrobium officinale roots and shoots, the increase of plant number and plant height, and the germination of new shoots and roots, which proved the effective establishment of symbiotic relationship between Tj2 strain and Dendrobium officinale. Therefore, T2 strain has practical application values on the successful cultivation of Dendrobium officinale plantlets.

A New Antioxidant Cyclic Diarylheptanoid from the Rhizomes of Zingiber officinale

The bioassay was directed to the fractionation of the methanol extract of the rhizomes of Zingiber officinale Roscoe leading to the isolation of a new antioxidant cyclic diarylheptanoid. The structure of the new compound was established as 1, 5-epoxy-3-hydroxy-1-(3,4-dihydroxy-5-methoxy-phenyl)-7-(3,4-dihydroxyphenyl) heptane (1) on the basis of MS, 1D and 2D-NMR experiments.

The Interplanting Technology of Pleurotus ostreatus and Dendrobium officinale

In summer, Cha No. 39 of 292 bags and 205 of 244 bags were inter- planted with Dendrobium officinale on cultivation bed, respectively, whose yields and biological efficiencies reached 102.46 kg and 103.98%, and 96.77 kg and 113.32%, accordingly. The seedlings of Dendrobium officinale interplanted with Pleurotus os- treatus grew by 2.78 cm during 10 weeks on cultivation beds, and the seedlings grew by 2.68 cm in the cultivation beds without interplanting arrangement. These demonstrated that the interplanting of Dendrobium officinale and Pleurotus ostreatus has none adverse effects on growth of Dendrobium officinale.

Study on Soil Nutrient Contents and Nutrient Characteristics of Ginger(Zingiber officinale Rosc.)

[Objective] This study was conducted to provide scientific basis for fertilization of ginger with high quality and high yield. [Method] The soil nutrient contents and plant nutrient characteristics of Zhugen ginger（Zingiber officinale Rosc.） were studied. [Result] The results showed that： the soil of ginger planting area in Chongqing（Yongchuan） Seedlings of science and technology city was acidic overall,the soil organic matter content was low and alkali-hydrolyzable N was at moderate level; and the contents of available P, Ca, Mg, Fe, Mn, Cu and Zn were extremely rich, while the available S was in lack. The contents of N and K were abundant and P was little in ginger plants, the content of K was more than N in ginger roots and stems, while the content of N was more than K in ginger leaves. [Conclusion]In practical production, it was necessary to fertilize rationally and control the ratio of N fertilizer to K fertilizer according to local conditions, thereby promoting ginger growth and realize high-yield ginger production.

Antibacterial effect of Allium sativum cloves and Zingiber officinale rhizomes against multiple-drug resistant clinical pathogens

Objective:To evaluate the antibacterial properties ot Allium sativum(garlic) cloves and Zingiber officinale(ginger) rhizomes against multi-drug resistant clinical pathogens causing nosocomial infection.Methods:The cloves of garlic and rhizomes of ginger were extracted with 95%(v/v) ethanol.The ethanolic extracts were subjected to antibacterial sensitivity test against clinical pathogens.Results:Anti-bacterial potentials of the extracts of two crude garlic cloves and ginger rhizomes were tested against five gram negative and two gram positive multi-drug resistant bacteria isolates.All the bacterial isolates were susceptible to crude extracts of both plants extracts.Except Enterobacter sp.and Klebsiella sp.,all other isolates were susceptible when subjected to ethanolic extracts of garlic and ginger.The highest inhibition zone was observed with garlic(19.4S mm) against Pseudomonas aeruginosa(P.aeruginosa).The minimal inhibitory concentration was as low as 67.00 μg/mL against P.aeruginosa.Conclusions:Natural spices of garlic and ginger possess effective anti-bacterial activity against multi-drug clinical pathogens and can be used for prevention of drug resistant microbial diseases and further evaluation is necessary.

Research Progress of Medicinal Secondary Metabolites and Gene Cloning of Dendrobium officinale

Dendrobium officinale is one of the most precious medicinal plants in China. Its main medicinal ingredients are its secondary metabolites. However,it has the characteristics of limited sources,low active ingredient content and high cost,limiting the use of D. officinale.Studying the network structure and rate-limiting steps of secondary metabolites of medicinal components of D. officinale,analyzing the secondary metabolic synthesis process,mastering the production rules of its medicinal components and carrying out gene cloning or biosynthesis,etc.are of great significance for the rational development and utilization of D. officinale resources. This paper briefly reviews the progress of the research on the secondary metabolites of D. officinale,including the detection and identification of metabolites and the identification and cloning of key metabolic enzymes.

Evaluation of androgenic activity of Zingiber officinale and Pentadiplandra brazzeana in male rats

Aim: Aqueous extracts of Zingiber officinale and Pentadiplandra brazzeana were tested for their possible androgenic activity in male Wistar rats. Methods: The aqueous extracts of the two plants were gavaged separately to 2 groups of rats at a similar dose of 600 mg.kg^1.day-1 for 8 days. At the end of the treatment, the animals were killed and the blood, testis, epididymis, seminal vesicles and prostate were collected for biochemical analysis. Results: The aqueous extract of Z. officinale significantly increased in the relative weight of the testis, the serum testosterone level, testicular cholesterol level and epididymal α-glucosidase activity. The aqueous extract of P. brazzeana significantly increased the weights of the testis, seminal vesicles and prostate. It also significantly increased the serum and testicular testosterone level. The fructose, α-glucosidase and cholesterol levels in P. brazzeanatreated rats were increased by 28%, 35% and 114%, respectively. Conclusion: The aqueous extracts of both P. brazzeana and Z. officinale have an androgenic activity, which seems to be more potent with P. brazzeana than with Z. officinale. (Asian J Androl 2002 Dec; 4: 299-301)

The protective effect of Dendrobium officinale polysaccharides on photoaging fibroblasts by scavenging reactive oxygen species and promoting the expression of TGF-β1

The purpose of this study was to explore the protective effect of Dendrobium officinale polysaccharides （DOP） onphotoaging human skin fibroblasts and its specific mechanism of action. The photoaging fibroblast model wasestablished by ultraviolet B （UVB） irradiation. The toxic effects of different concentrations of DOP were detected usingMTT. Senescent cells were detected using a β-galactosidase kit. Reactive oxygen species （ROS） in cells were detectedusing a flow cytometer. The expression of matrix metalloproteinase-1 （MMP-1）, type I collagen C-terminal peptide（CICP）, and transforming growth factor β-1 （TGF-β1） in spent culture medium was detected by ELISA. The resultsshowed that the low concentration of DOP （20, 40, 80 μg/mL） had no cytotoxicity on fibroblasts. After 60 mJ/cm2UVBirradiation, the number of aging β-gal-positive cells increased, the levels of CICP and TGF-β1 in spent culture mediumdecreased, while the levels of MMP-1 and ROS increased. After administration of DOP on photoaging fibroblasts, thenumber of aging β-gal-positive cells decreased, the levels of ROS and MMP-1 decreased, and the levels of TGF-β1 andCICP increased. This experiment suggests that DOP has the effect of removing ROS induced by UVB, regulating thebalance of collagen production and degradation, and protecting photoaging human skin fibroblasts.

Two new compounds from Zingiber officinale

A new cyclic diarylheptanoid, 1,5-epoxy-3-hydroxy-1-（3-methoxy-4,5-dihydroxyphenyl）-7-（4-hydroxyphenyl）-heptane （1）, as well as a new monoterpene, 10-O-β-n-glucopyranosyl-hydroxy cineole （2） were isolated from the rhizomes of Zingiber officinale. The structures of compounds 1 and 2 were established based on their spectral data. In addition, the antioxidant activities of these compounds were also measured.

Densitometric HPTLC analysis of 8-gingerol in Zingiber officinale extract and ginger-containing dietary supplements,teas and commercial creams

Objective:To develop and validate a simple,accurate HPTLC method for the analysis of 8-gingerol and to determine the quantity of 8-gingerol inZingiber officinaleextract and gingercontaining dietary supplements,teas and commercial creams.Methods:The analysis was performed on 10×20 cm aluminium-backed plates coated with 0.2 mm layers of silica gel 60 F254(E-Merck,Germany)with n-hexane:ethyl acetate 60:40(v/v)as mobile phase.Camag TLC Scanner III was used for the UV densitometric scanning at 569.Results:This system was found to give a compact spot of 8-gingerol at retention factor(Rf) value of(0.39依0.04)and linearity was found in the ranges 50-500 ng/spot(r2=0.9987).Limit of detection(12.76 ng/spot),limit of quantification(26.32 ng/spot),accuracy(less than 2%)and recovery(ranging from 98.22-99.20)were found satisfactory.Conclusions:The HPTLC method developed for quantification of 8-gingerol was found to be simple,accurate,reproducible,sensitive and is applicable to the analysis of 8-gingerol in Zingiber officinaleextract and ginger-containing dietary supplements,teas and commercial creams.

Two New Diarylheptanoids from the Rhizomes of Zingiber officinale

Two new diaryheptanoids, (5S)-5-acetoxy-1,7-bis(4-hydroxy-3-methoxyphenyl)-3- heptanone (1) and (3S,5S)-3,5-diacetoxy-1,7-bis(4-hydroxy-3-methoxyphenyl)heptane (2) were isolated from the rhizomes of Zingiber officinale. Their structures were elucidated by spectral methods.

Investigation on the Morphological Characteristics of Dendrobium officinale Plantlets Propagated from Different Explants

[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale.

Evaluation of Heavy Metals and Pesticide Residues in Dendrobium officinale Produced in Guangxi

[Objectives]To establish a method for the determination of heavy metals and pesticide residues in Dendrobium officinale produced in Guangxi and its culture substrate.[Methods]The contents of five heavy metals(Pb,Cd,As,Hg and Cu)were determined by the atomic absorption spectrophotometric(AAS)method;residues of 9 organochlorine pesticides(α-BHC,β-BHC,γ-BHC,δ-BHC,p,p′-DDT,p,p′-DDE,p,p′-DDD,o,p′-DDT and pentachloronitrobenzene)were determined by gas chromatography(GC).[Results]The contents of five heavy metals in the medicinal materials of D.officinale produced in Guangxi were 0.96,0.21,0.021 mg/kg,and not detected,and 0.35 mg/kg.The contents of five heavy metals in the culture substrate were 0.03,0.19,0.21,0.004 and 1.80 mg/kg,respectively;the content of 9 organochlorine pesticide residues in the sample was not detected.[Conclusions]The detection results of heavy metal content and organochlorine pesticide residues in D.officinale produced in Guangxi and its substrate are consistent with the limit requirements of the Chinese Pharmacopoeia.This experiment is expected to provide reliable data reference for ensuring the safety of the drug resources of D.officinale in Guangxi.

The bibenzyl derivatives of Dendrobium officinale prevent UV-B irradiation induced photoaging via SIRT3

Dendrobium officinale is a valuable medicinal herb that is widely used in traditional Chinese medicine.The chemical constituents of D.officinale have attracted much attention and a large number of compounds have been reported including many bibenzyl derivatives.13 bibenzyl derivatives from D.officinale were sent for molecular docking,sur-face plasmon resonance(SPR)assay and after detection of Mn-SOD and SIRT3 activities in or not in HaCaT cells,it was concluded that bibenzyl derivatives did not directly activate Mn-SOD but promoted SIRT3 proteins.In addition,HaCaT cells were irradiated with UV-B to induce an oxidative stress model in vitro to further verify the effect of bibenzyl derivatives.The results show that bibenzyl derivatives could directly bind to SIRT3,enhance the deacetylation and then activate Mn-SOD,so as to protect UV-B induced skin photoaging.

Optimization of Rooting Medium for Tissue Culture Seedlings of Dendrobium officinale by Response Surface Methodology

[Objectives] To increase the survival rate of tissue culture seedlings of Dendrobium officinale,and optimize the conditions of rooting medium by the response surface methodology( RSM). [Methods]The effects of 6-BA concentration,NAA concentration,potato amount and the amount of mashed banana on the growth of seedlings were determined by single factor experiment and were analyzed by Box-Behnken design and response surface methodology. [Results]The optimal culture conditions: rooting medium 1/2 MS + 6-BA 0. 24 mg/L + mashed banana 87. 63 g/L + potato 89. 30 g/L + NAA 0. 52 mg/L + sucrose 20. 0 g/L + activated carbon 4. 0 g/L + agar 7. 0 g/L,p H 5. 8,and light intensity 2 000 Lx. [Conclusions]The model established by response surface methodology has a good predictability and could be used to optimize the conditions of tissue culture and rooting medium of D. officinale.

Physiological Responses of Dendrobium officinale under Exposure to Cold Stress with Two Cultivars

This study aimed to explore the cold tolerance of two cultivars of Dendrobium officinale(MG1,MG2)grown in different regions of China.Under-2℃ incubation,cultivar MG1 remained active after 3 d,and continued to grow after returning to room temperature.However,MG2 could only maintain its activity after 2 d treatment at−2℃,and the seedlings died with the low temperature treatment time.Investigation of the characteristics of the plants grown in the south(Hangzhou)or north(Zhengzhou)of China indicated that the leaves of MG1 also had reduced stomatal density,the highest thickness,and a compact microstructure.The contents of proline and soluble sugars were higher in MG1 than those in MG2.The cultivar MG1 had higher SOD enzyme activity than MG2,while CAT and POD activities in samples from Zhengzhou were higher than those from Hangzhou.The contents of polysaccharides and alkaloids in stems of in MG1 were higher than those in MG2,while the content of flavonoids in the Zhengzhou samples was higher than that in the Hangzhou samples.In addition,plant heights,stem diameters,and chlorophyll content were higher in MG1.Overall,MG1 had better cold resistance than MG2.MG1 is a cold tolerant cultivar with thick leaves and reduced stomatal density,higher contents of soluble sugars,proline,CAT,POD,polysaccharides,flavonoids and alkaloids,which together make it more adaptable to low temperatures.Thus,the cultivar MG1,with its demonstrated cold tolerance,can accordingly be grown on a large scale in cold regions,thereby expanding the available planting area for this important traditional medicinal plant to meet the increasing commercial demand for it.