ABSTRACT
This study aims to explore the relationship of DNA methylation with the contents of the index components as well as the growth and development of Pogostemon cablin. The demethylation reagent 5-azacytidine(5-azaC) was used to treat the tissue culture seedlings of patchouliol-type P. cablin. High performance liquid chromatography was employed to evaluate the changes of DNA methy-lation in P. cablin, and GC-MS to detect the contents of index components in P.cablin. The agronomic characters of P.cablin were measured using the common methods. The results showcased that DNA methylation of P.cablin was significantly reduced by 5-azaC in a concentration-dependent manner. Thirty days after treatment with 5-azaC at different concentrations, the content of patchouli alcohol changed slightly; compared with that in the control group, the content of pogostone in 50 μmol·L~(-1) and 100 μmol·L~(-1) 5-azaC groups was significantly up-regulated. The 100 μmol·L~(-1) 5-azaC group had the largest differences in contents of pogostone and patchouli alcohol compared with the control group, followed by the 50 μmol·L~(-1) 5-azaC group. Ninety days after disinhibition, the content of pogostone in the treatment group was significantly increased and the content of patchouli alcohol was significantly decreased. In addition, 5-azaC significantly inhibited the growth and development of P.cablin in a dose-dependent manner. These results indicate that DNA methylation regulates the biosynthesis of the index components in patchouliol-type P.cablin and proper demethylation can directly promote the synthesis of pogostone and indirectly affect the accumulation of patchouli alcohol.
Subject(s)
Azacitidine , DNA Methylation , Gas Chromatography-Mass Spectrometry , Oils, Volatile , Pogostemon/geneticsABSTRACT
The terpenoids in Pogostemon cablin have complex structures and abundant pharmacological effects. Patchouli alcohol(PA) and pogostone(PO) have a high medicinal value by virtue of anti-tumor, anti-inflammatory, antibacterial, antioxidant, and other biological activities. Due to the low content of terpenoid metabolites in P. cablin, the study of biosynthesis and metabolism regulation can provide a biosynthetic basis for obtaining high-content terpenoids. In this study, key enzyme genes in biosynthesis, transcription factors in metabolism regulation, spatio-temporal expression of terpene synthase were reviewed, aiming to provide a reference for the development, protection, and utilization of P. cablin resources.
Subject(s)
Pogostemon/genetics , Terpenes , Transcription Factors/geneticsABSTRACT
OBJECTIVE@#To investigate the antibacterial activity of patchouli alcohol (PA) against 127 bacteria strains, including the common bacteria and drug-resistant bacteria strains both in the in vitro and in vivo tests.@*METHODS@#For the in vitro trial, the antibacterial property of PA against 107 Gram-positive and 20 Gram-negative bacteria strains was screened by agar double dilution method. For the in vivo trial, specific pathogen free Kunming strain of both male and female white mice, were used to test the protective ability of PA after being injected with the median lethal dose of the tested strains.@*RESULTS@#PA possessed antibacterial activity against all the tested 127 strains. In the in vitro test, PA could inhibit both Gram-negative bacteria (25-768μg/mL) and Gram-positive bacteria (1.5-200μg/mL). Particularly, PA was active against some drug-resistant bacteria like methicillin-resistant Staphylococcus aureus (MRSA). PA also exhibited in vivo anti-MRSA activity in mice via intraperitoneal injection. PA could protect mice entirely infected with MRSA at 100 and 200 mg/kg, while 80% mice injected with MRSA could be protected at a low dose of 50μg/mL.@*CONCLUSION@#PA might be a potential antibacterial drug from natural sources and might be worthy to explore its mechanism and application in further study.
ABSTRACT
Patchouli Alcohol (PA) is the main bioactive constituent of Pogostemon cablin Pharmacological researches have reported various benefits of PA, including anti-infammatory, anti-gastric ulcer, anticancer, protecting against alung and brain injury, antibacterial, prevention of mastitis, colitis and arteriosclerosis, anti-photoaging, treatment of diarrhea irritable bowel syndrome, antioxidant stress, vasodilation and anti-injury.The preliminary study on the mechanism showed that anti-inflammatory effect was related to the down-regulation of mRNA expression of inflammatory mediators to inhibit the production of proinflammatory cytokines and the reduction of extracellular regulated protein kinases (ERK)-mediated nuclear factor kappa B (NF-κB) activation.The anti-gastric ulcer, protection of lung injury and brain injury and anti-photoaging were related to antioxidant and anti-inflammatory effects.The mechanism of anti-human leukemic cell line MV4-11 migh be associated with the changes of NF-κB, phosphorylated M2 pyruvate kinase (p-PKM2) and cysteine aspartate protease-3 (Caspase-3) protein expressions.The proposed mechanisms of anti-human colorectal cancer cells included the inhibition of histone deacetylase (HDAC)-2 expression and HDAC enzyme activity, and subsequent downregulation of c-myc and activation of NF-κB pathway.Apoptosis of A549 cells was induced by blocking the phosphorylation of epidermal growth factor receptor (EGFR) pathway and activating c-Jun N-terminal kinase (JNK). Anti-prostate cancer was related to the activation of mitochondrial cytochrome C apoptosis pathway, and its antiviral effect may be related to the regulation of RIG-1-like helicases (RLH) signal pathway or the direct binding of virus protein or nucleic acid and the binding site of virus to cells to block the adsorption of virus to cells.The mechanism of its antibacterial, prevention of mastitis, colitis and arteriosclerosis was closely related to anti-inflammation.The pharmacological effects of PA need to be further studied.In order to accelerate the exploration and discovery other molecular mechanism of PA, and expand its application prospects.This review comprehensively summarizes the metabolism, biological activities, and possible mechanism of PA, in order to provide references for the development and application of PA.
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Objective::To investigate the pharmacodynamics of volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba, to establish a method for simultaneous determination of three essential ingredients (thymol, carvacrol and patchouli alcohol) in volatile oil of the couplet medicines by gas chromatography (GC), to optimize the preparation process of β-cyclodextrin (β-CD) inclusion complex of volatile oil in the couplet medicines and to confirm the formation of the inclusion complex. Method::An in vitro inflammatory response model was established by hyaluronidase activity inhibition test in order to detect the anti-inflammatory activity of the volatile oil. Also, the antioxidant activity of the volatile oil was assessed by 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical scavenging method. The inclusion of volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba was prepared by scaturated aqueous solution method, colloid milling method and grinding method, respectively. GC was used to determine the contents of thymol, carvacrol and patchouli alcohol in volatile oil for optimizing extraction and inclusion processes of volatile oil. Scanning electron microscope, infrared spectroscopy, thermal differential analysis, and X-ray diffraction (XRD) were used to verify the formation of the inclusion complex. Result::The volatile oil not only inhibited hyaluronidase activity to a certain extent, but also eliminated DPPH and increased with the increase of concentration. There was a good linear relationship between the peak area and concentration of thymol, carvacrol and patchouli alcohol at 0.021 3-0.426, 0.020 04-0.400 8, 0.022 6-0.452 g·L-1 (R2>0.999), respectively. Their recoveries were 99.59%(RSD 1.6%), 100.15%(RSD 1.5%), 100.70%(RSD 1.4%), respectively. The colloid milling method was optimized, and the formation of the inclusion complex was verified by the aforementioned methods. Conclusion::The volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba has certain anti-inflammatory activity and anti-oxidation ability. The colloid milling method was the best inclusion process for the volatile oil. The established GC has the advantages of simple, sensitive, accurate, reliable and reproducible, which can meet the requirements of simultaneous determination of thymol, carvacrol and patchouli alcohol in the inclusion complex.
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OBJECTIVE@#To explore the anti-nociceptive effect of patchouli alcohol (PA), the essential oil isolated from Pogostemon cablin (Blanco) Bent, and determine the mechanism in molecular levels.@*METHODS@#The acetic acid-induced writhing test and formalin-induced plantar injection test in mice were employed to confirm the effect in vivo. Intracellular calcium ion was imaged to verify PA on mu-opioid receptor (MOR). Cyclooxygenase 2 (COX2) and MOR of mouse brain were expressed for determination of PA's target. Cellular experiments were carried out to find out COX2 and MOR expression induced by PA.@*RESULTS@#PA significantly reduced latency period of visceral pain and writhing induced by acetic acid saline solution (P<0.01) and allodynia after intra-plantar formalin (P<0.01) in mice. PA also up-regulated COX2 mRNA and protein (P<0.05) with a down-regulation of MOR (P<0.05) both in in vivo and in vitro experiments, which devote to the analgesic effect of PA. A decrease in the intracellular calcium level (P<0.05) induced by PA may play an important role in its anti-nociceptive effect. PA showed the characters of enhancing the MOR expression and reducing the intracellular calcium ion similar to opioid effect.@*CONCLUSIONS@#Both COX2 and MOR are involved in the mechanism of PA's anti-nociceptive effect, and the up-regulation of the receptor expression and the inhibition of intracellular calcium are a new perspective to PA's effect on MOR.
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Objective: To investigate the ultrasound-assisted extraction process of Pogostemon cablin. Methods: Taking the extraction time (min), ethanol concentration (％), and liquid-solid ratio (mL · g-1) as influence factors, patchouli alcohol (PA) content as index, single-factor test was applied to the investigation of extraction-process of Pogostemon cablin. By ultrasonic-assisted extraction, the GC-FID was used to determine the PA content in extract. Results: The optimal extraction conditions were as follows: the ultrasonic time was 30 min, the ethanol concentration was 95％, and liquid-solid ratio was 20∶1 mL·g-1. Under this condition, the extraction rate and the content of PA were 32.75％, and0.15％ respectively. Conclusion: This extraction method is simple, but has higher extraction rate. It is suitable for the industrial production.
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The aim of this paper is to investigate the effect of patchouli alcohol in enhancing Helicobater pylori's action in eradicating macrophages and its mechanism. H. pylori was co-cultured with macrophages at a ratio of MOI=100 in different concentrations of patchouli alcohol. The effect of patchouli alcohol in eradicating macrophages was detected by agar dilution method. The effect of patchouli alcohol on NO and myeloperoxidase (MPO) levels in macrophages were measured by H. pylori by biochemical methods. Patchouli alcohol effect on H. pylori-induced pro-inflammatory gene expression and protein secretion in macrophages were detected by RT-qPCR and ELISA method. The eradication of H. pylori has significantly enhanced, and the destabilization of lysosomes has been reversed. Meanwhile, patchouli alcohol has an effect in inhibiting pro-inflammation and oxidation. The mechanism of patchouli alcohol in eradicating H. pylori and resisting oxidative stress may be associated to the blocking of bacteria escape lysosome combination procedures.
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Objective To analytically screen the volatile components from Chenxiang Huaqi Tablets (CHT) using gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) combined with automated mass spectral deconvolution, and establish the methods of multiple components contents (camphor, bornyl acetate, and patchouli alcohol) from CHT using multi reaction monitor (MRM). Methods The chromatographic column was HP-5MS (30 m × 0.32 mm × 0.25 μm). Temperature program: the initial temperature was 100 ℃, at 15 ℃/min up to 300 ℃ for 2 min. Injection port temperature was 280 ℃; Ionization methods: EI ion source with ion source temperature of 230 ℃; Quadrupole temperature was 150 ℃; Interface temperature was 280 ℃; Inject volume was 1 μl. Results A total of thirteen components were screened by automated mass spectral deconvolution soft. Under the conditions of MRM collection, camphor, bornyl acetate, and patchouli alcohol were in the range of 4.5-90 mg/mL (r = 0.999 8), 3.3-66 mg/mL (r = 0.999 8), and 2.6-51.5 mg/mL (r = 0.999 9), respectively, the ratio of each component concentration to its peak area was linear, and the average recovery (n = 6) was 101.15%, 102.64%, and 100.10% respectively; The mass fraction of 10 batches of samples were in the ranges of 0.278-0.311, 0.381-0.438, and 0.229-0.381. Conclusion The method is accurate, simple, and good repeatability. It can be used for the simultaneous determination of camphor, bornyl acetate, and patchouli alcohol in CHT, which provides a reference for the improvement of the quality of this variety.
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Cancer is a serious threat to human health with a high fatality rate. Modern anticancer methods, including surgery, radiotherapy, and chemotherapy, commonly have more serious side effects. Anticancer plant essential oils which now attach great importance to cancer treatment, have many advantages such as multi-target, multi-effect, low adverse reaction, improving the body immunity, not easy to produce drug resistance, etc. The main active ingredients and the anticancer mechanism of essential oils were reviewed in this paper. Nineteen kinds of plant essential oils, including Cymbopogon flexuosus, Salvia officinalis, Lycopus lucidus, Lavandula angustifolia, Smyrnium olusatrum, peel of Citrus reticulate, leaves of Myrica rubra, and so on, displayed their prominent anti-cancer activities. Specifically, limonene, β-elemene, menthol, patchouli alcohol, thymol, eugenol, citral, β-caryophyllene, and their oxide may be the main anticancer composition of plant essential oils.
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To investigate the effect of patchouli alcohol on inhibiting Helicobater pylori urease activity, and its effect on expression levels of related genes, and lay the foundation for further research on the effect of patchouli alcohol on H. pylori colonization and infection. H. pyloriwas cultured and identified by gram staining, rapid urease test (RUT) and PCR method. Then agar dilution method was used to detect the bacterial survival after 1 h intervention by different concentrations of patchouli alcoholin the acidic (pH 5.3) and neutral (pH 7.0) conditions; berthelot method was used to detect urease activity and RT-qPCR method was used to detect the expression changes of ureA, ureB, ureE, ureH, ureI, and nixA related urease genes. The results showed that the survival rate of H. pyloriwas not significantly changed but the urease activity was obviously decreased after intervention by different concentrations of patchouli alcohol; meanwhile, the expression levels of ureA, ureB, ureE, ureH, ureI, and nixA were decreased to different degrees. Therefore, patchouli alcohol could inhibit H. pylori urease activity in both acidic and neutral conditions, and the mechanism may be related to down-regulation of urease gene expression.
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Objective:To develop a method for the quality control of Cuochuang powder .Methods:Rhei Radix et Rhizoma, An-gelicae Dahuricae Dadix and Saposhnikoviae Radix were identified by TLC .GC was used for the determination of patchouli alcohol , menthol and borneol .Results:The TLC spots were clear without any interference from the negative control .The linear range of pat-chouli alcohol was 0.020 1-0.805 6 mg· ml-1 , that of borneol was 0.010 0-0.401 6 mg· ml-1 , and that of menthol was 0.005 1-0.202 8 mg· ml-1.The average recovery (n=6) was 102.03% (RSD=0.91%), 100.10% (RSD =1.94%) and 103.15%(RSD=1.68%),respectively.Conclusion:The method is simple, accurate and repeatable, which can be used for the quality control of Cuochuang powder .
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OBJECTIVE:To establish a method for the contents determination of volatile components in Huodan pill and study the effects of 60Co irradiation with different doses on the volatile components in Huodan pill. METHODS:GC-MS was used to de-termine the contents of patchouli alcohol and the ralative contents of β-patchouliene,α-guaiacene,seychellesene,α-patchouliene,α-bulnesene,5,11-diene guaiacyl,and unidentified objects in Huodan pill after 60Co irradiation with different doses. Column was HP-5MS by programmed temperature,volume temperature was 250℃,split ratio was 50:1,detector was triple quadrupole mass spectrometer detector,MS1 quadrupole temperature was 150℃,MS2 was 150℃,carrier gas was helium,flow rate of column was 1.2 ml/min,and the volume injection was 1 μl;ion source was EI,bombarding energy was 70 eV,temperature of ion source was 230℃,temperature of transmission line was 280℃ and scan range was 50-500 amu. RESULTS:The linear range of patchouli alcohol was 0.103 1-2.062 0μg(r=0.999 4);RSDs of precision,reproducibility and stability tests were lower than 1.0%;recovery was 98.05%-102.32%(RSD=1.8%,n=9). 60Co irradiation with different doses had little effects on the 8 volatile components in Huodan pill. CONCLUSIONS:60Co irradiation can be used for sterilization on Huodan pill,but irradiation dose should be controlled. The method is simple,good reproducibility,and can be used for the contents determination of volatile components in Huodan pill.
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Sesquiterpenes Essential oil produced by patchouli was one of the most important naturally occurring base materials used in the perfume industry, containing various sesquiterpenes. Three different parts (leaves, stems and roots) of Pogostemon cablin (Blanco) Benth., Lamiaceae, were profiled in relation to different maturation phases in this paper, evaluating the variations in content of the major sesquiterpenes in the essential oil. Twelve sesquiterpenes were analyzed by GC-MS throughout the maturity of P. cablin. Patchouli alcohol (37.54%-51.02% in leaves, 28.24%-41.96% in stems and 14.55%-35.12% in roots) was the major sesquiterpene during the maturation of the plant. The average content of several other sesquiterpenes (α-bulnesene, α-guaiene, seychellene, β-humulene and caryophyllene) were higher than 3% among leaves, stems and roots. The content of essential oil, patchouli alcohol, α-bulnesene and several other compounds were highly accumulated at 210 days of maturation after cultivation of P. cablin. Thus, this period was the best moment to exploit the maximum level of these high value-added compounds in P. cablin. Furthermore, our results indicated that the essential oil extracted from leaves of P. cablin has the highest potential to be used in the perfume industry.
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OBJECTIVE: To determine the content of patchouli alcohol in Huodan pills (traditional Chinese medicine) by near-infrared (NIR) spectroscopy. METHODS: The content of patchouli alcohol in 147 batches of Huodan pills samples were determined by GC. NIR spectra of all samples were collected, then standard normal variate, first derivative and Norris derivative filter methods were used as spectral preprocessing tools, and at last, the calibration model of patchouli alcohol content was established by partial least squares regression. RESULTS: The correlation coefficients, the root-mean-square error of calibration, and the root-mean-square error of cross-validation of the calibration model for patchouli alcohol were 0.95509, 0.0315, and 0.04510, respectively; the root-mean-square error of prediction and the average recovery rate of validation samples were 0.0343 and 100.76%, respectively. CONCLUSION: The method is stable, accurate, and can be used to determine the content of patchouli alcohol in Huodan pills rapidly.
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OBJECTIVE:To determine the content of Patchouli alcohol in Xiaoer tuxiening powder by gas chromatography(GC).METHODS:Using n-octadecane as internal standard,the determination was performed by the GC on elastic quartz ca-pillary column(30 m?0.32 mm?0.25 ?m)with 5% phenyl-methylpolysiloxane as stationary phase that was heated by temperature programming with both detector temperature and injector temperature maintained at 280 ℃.RESULTS:The linear range of Patchouli alcohol was 63.4～887.6 ?g?mL-1(r=0.999 4),and its average recovery rate was 100.55%(RSD=1.85%,n=6).CONCLUSION:The GC method has been proved to be rapid,accurate and reliable,and it is applicable for the quality control of Xiaoer tuxiening powder.
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Objective The volatile oil was further purified to get the patchouli alcohol by molecular distillation taking patchouli oil which contained 23.97% of patchouli alcohol as raw material.Methods The best process parameters,including temperature,feed rate,and scraping rate,of molecular distillation technique for separating patchouli oil were studied by orthogonal test.The content was detected by GC under the conditions of elastic quartz capillary column HP-5,detector temperature 280 ℃,gasification temperature 280 ℃,distributary refer 20∶1.The effects of evaporation temperature on content and extraction rate of the patchouli oil had been investigated and the difference between the molecular and vapor distillations was studied.Results Content and extraction rate of patchouli alcohol under the best evaporation temperature 65 ℃ by molecular distillation reached to 40.71% and 76.55%,respectively.Conclusion The molecular distillation technique that could get the product with higher purity and extracting rate than that by vapor distillation in patchouli oil purification has a very good prospect in industry.
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Objective To establish a method for the determination of patchouli alcohol in Huoxiang Qingwei Tablets by gas chromatography (GC).Methods HP- 5 column( 30 m? 0.32 mm? 0.25 ? m) was used and 5 % crosslinked phenyl polydimethyl siloxane was used as the mobile phase. The increase of column temperature was controlled by programming: the initial temperature was 150 ℃ and maintained for 23 min, and then rose at 8 ℃ /min up to 230 ℃ for 2 min. The injector temperature and the detector temperature both were 280 ℃ .The flow rate was 0.9 mL/min and the injection volume was 1 ? L, split ratio was 5∶ 1.Result A good linearity was obtained in the range of 0.057～ 1.140 mg/mL,r=0.9994. The average recovery was 99.38 % with a RSD of 1.82 % (n=5). Conclusion The method is simple and accurate and can be used for the quality control of Huoxiang Qingwei Tablets.