Effects of processing technology on essential oil and pharmacological action of frankincense

Background:According to the traditional Chinese medicine theory,the common processing methods for Ruxiang(frankincense)include stir-frying processing and vinegar processing.Methods:With network pharmacology and thermal analysis methods,we selected and identified the main and irritating ingredients,established the pyrolysis characteristic parameters for identifying frankincense quality,quantified the starting and optimum temperature limits for processing frankincense,and analyzed the influences of processing adjuvants(vinegar)on combustion pyrolysis characteristics of frankincense.By applying Fourier transformation infrared spectrometry,high performance liquid chromatography,gas chromatography and UV-visible spectrometry,we evaluated the processing technology developed in our study of frankincense and its processed products.Results:Based on network pharmacology,we can find that the classical compounds of essential oil are the main pharmacodynamics components of frankincense possessing the anti-inflammatory,analgesia,anti-tumor,anti-ulcer and ant-inflammatory bowel disease effects.The pyrolysis combustion rate peaks of frankincense at 285.27±7.05℃and 476.99±13.46℃were the characteristic peaks used to identify frankincense quality.The specific temperature of slow-fire processing,including the vinegar processing and stir-frying processing,was 148.72±5.09℃.Heating for 5.67±0.98 min after reaching processing temperature was needed for processing frankincense with vinegar.The kinetics analysis showed that both frankincense and its vinegar mixture exhibited good linear relationships.The feasibility and practicability for our processing technology were verified through the determination of medicinal compounds by Fourier transformation infrared spectrometry,high performance liquid chromatography,gas chromatography and UV.Conclusion:Frankincense essential oils were the main active components of frankincense,which also can be taken as the internal chemical control indexes to judge the quality control and to optimize the processing technology of frankincense.The pyrolysis combustion rate peaks of frankincense at 285.27±7.05℃and 476.99±13.46℃were the characteristic peaks that can be used to identify frankincense quality.The optimum processing conditions of vinegar-processed frankincense with were as follows:with vinegar,20%;heating temperature,148.72±5.09℃;and heating time,5.67±0.98 min.Under these conditions,frankincense essential oil could cause a moderate effect on combustion pyrolysis characteristics to exert its corresponding pharmacological effects.

Frankincense myrrh attenuates hepatocellular carcinoma by regulating tumor blood vessel development through multiple epidermal growth factor receptor-mediated signaling pathways

BACKGROUND In traditional Chinese medicine(TCM),frankincense and myrrh are the main components of the antitumor drug Xihuang Pill.These compounds show anticancer activity in other biological systems.However,whether frankincense and/or myrrh can inhibit the occurrence of hepatocellular carcinoma(HCC)is unknown,and the potential molecular mechanism(s)has not yet been determined.AIM To predict and determine latent anti-HCC therapeutic targets and molecular mechanisms of frankincense and myrrh in vivo.METHODS In the present study,which was based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(http://tcmspw.com/tcmsp.php),Universal Protein database(http://www.uniprot.org),GeneCards:The Human Gene Database(http://www.genecards.org/)and Comparative Toxicogenomics Database(http://www.ctdbase.org/),the efficacy of and mechanism by which frankincense and myrrh act as anti-HCC compounds were predicted.The core prediction targets were screened by molecular docking.In vivo,SMMC-7721 human liver cancer cells were transplanted as xenografts into nude mice to establish a subcutaneous tumor model,and two doses of frankincense plus myrrh or one dose of an EGFR inhibitor was administered to these mice continuously for 14 d.The tumors were collected and evaluated:the tumor volume and growth rate were gauged to evaluate tumor growth;hematoxylineosin staining was performed to estimate histopathological changes;immunofluorescence(IF)was performed to detect the expression of CD31,α-SMA and collagen IV;transmission electron microscopy(TEM)was conducted to observe the morphological structure of vascular cells;enzyme-linked immunosorbent assay(ELISA)was performed to measure the levels of secreted HIF-1αand TNF-α;reverse transcription-polymerase chain reaction(RT-qPCR)was performed to measure the mRNA expression of HIF-1α,TNF-α,VEGF and MMP-9;and Western blot(WB)was performed to determine the levels of proteins expressed in the EGFR-mediated PI3K/Akt and MAPK signaling pathways.RESULTS The results of the network pharmacology analysis showed that there were 35 active components in the frankincense and myrrh extracts targeting 151 key targets.The molecular docking analysis showed that both boswellic acid and stigmasterol showed strong affinity for the targets,with the greatest affinity for EGFR.Frankincense and myrrh treatment may play a role in the treatment of HCC by regulating hypoxia responses and vascular system-related pathological processes,such as cytokine-receptor binding,and pathways,such as those involving serine/threonine protein kinase complexes and MAPK,HIF-1 and ErbB signaling cascades.The animal experiment results were verified.First,we found that,through frankincense and/or myrrh treatment,the volume of subcutaneously transplanted HCC tumors was significantly reduced,and the pathological morphology was attenuated.Then,IF and TEM showed that frankincense and/or myrrh treatment reduced CD31 and collagen IV expression,increased the coverage of perivascular cells,tightened the connection between cells,and improved the shape of blood vessels.In addition,ELISA,RT-qPCR and WB analyses showed that frankincense and/or myrrh treatment inhibited the levels of hypoxia-inducible factors,inflammatory factors and angiogenesis-related factors,namely,HIF-1α,TNF-α,VEGF and MMP-9.Furthermore,mechanistic experiments illustrated that the effect of frankincense plus myrrh treatment was similar to that of an EGFR inhibitor with regard to controlling EGFR activation,thereby inhibiting the phosphorylation activity of its downstream targets:the PI3K/Akt and MAPK(ERK,p38 and JNK)pathways.CONCLUSION In summary,frankincense and myrrh treatment targets tumor blood vessels to exert anti-HCC effects via EGFR-activated PI3K/Akt and MAPK signaling pathways,highlighting the potential of this dual TCM compound as an anti-HCC candidate.

Frankincense,a Special Spice Walking Along the Silk Road

Frankincense is also called“Fumigated land,”“Horsetail spice,”and“Godsend spice.”The alias“Fumigated land”shows its strong fragrance that can cover the whole land.The name of“Horsetail spice”implies its morphologic characteristics and mild effects,and another name of“Godsend spice”indicates its functions of relieving swelling and pain,healing sore,and growing muscle,as if it is the grace donated by God.The name of“Frankincense”displays its complex productive progress.With thick quality and strong fragrance,frankincense acts as daily supplies for religious activities and folk activities due to its low price and a wide range of uses in medicine and daily life.Frankincense shows the infinite charm of spice culture along the Silk Road from the distant Western Regions to the East.At the same time,it enriches the treasure house of traditional Chinese medicine and becomes one of the most popular spices in both eastern and western,nobility and common.

Efficacy of Frankincense and Myrrha in Treatment of Acute Interstitial Cystitis/Painful Bladder Syndrome

Objective:To investigate the efficacy of frankincense and myrrha in the treatment of acute interstitial cystitis/painful bladder syndrome(IC/PBS).Methods:The effects of frankincense and myrrha on the proliferation and migration of primary human urothelial cells(HUCs)were assessed in vitro.In the animal study,48 virgin female rats were randomized into 4 groups(12 in each group):(1)control group(saline-injected control);(2)cyclophosphamide(CYP)group(intraperitoneal injected 150 mg/kg CYP);(3)CYP+pentosan polysulfate sodium group(orally received 50 mg/kg pentosan polysulfate sodium);and(4)CYP+frankincense and myrrha group[orally received frankincense(200 mg/kg)and myrrha(200 mg/kg)].Rats orally received pentosan polysulfate sodium or frankincense and myrrha on day 1,2,and 3.The experiments were performed on day 4.Pain and cystometry assessment behavior test were performed.Voiding interval values were assessed in rats under anesthesia.Finally,immunohistochemistry and Western blot were used to confirm the location and level,respectively,of cell junction-associated protein zonula occludens-2(ZO-2)expression.Results:Low dose frankincense and myrrha increased cell proliferation and migration in HUCs compared with control(P<0.05).Rats with acute IC/PBS rats exhibited lower voiding interval values,pain tolerance,and ZO-2 expression(P<0.05).Voiding interval values and pain tolerance were higher in the frankincense and myrrha group than CYP group(P<0.05).ZO-2 expression in the bladder was increased in the CYP+pentosan polysulfate and frankincense+myrrha groups compared with the CYP-induced acute IC/PBS group(P<0.05).Conclusion:frankincense and myrrha modulate urothelial wound healing,which ameliorates typical features of acute IC/PBS in rats.

Geographical Origin and Level Identification of Frankincense Based on Hyperspectral Image

Background:As the demand for traditional Chinese medicinal materials increases in China and even the world,there is an urgent need for an effective and simple identification technology to identify the origin and quality of the latter and ensure the safety of clinical medication.Mineral element analysis and isotope finger-printing are the two commonly used techniques in traditional origin identification.Both of these techniques require the use of stoichiometric methods in the identification process.Although they have high accuracy and sensitivity,they are expensive and inefficient.In addition,near-infrared spectroscopy is a fast,nondestructive,and widely used identification technique developed in recent years,but its identification results are susceptible to samples’states and environmental conditions,and its sensitivity is low.Hyperspectral imaging combines the advantages of imaging technology and optical technology,which can simultaneously access the image information and spectral information which reflect the external characteristics,internal physical structure,and chemical composition of the samples.Hyperspectral imaging is widely applied to agricultural product inspection,but research into its application in origin and quality identification of TCM materials is rare.Methods:In this study,the algorithm framework discriminative marginalized least squares regression(DMLSR)was used for feature extraction of frankincense hyperspectral data.The DMLSR with intraclass compactness graph and manifold regularization can efficiently learn the projective samples with higher separability and less redundant information than the original samples.Then,the discriminative collaborative representation with Tikhonov regularization(DCRT)was applied for classifying the geographical origin and level of frankincense.DCRT introduces the discriminant regularization term and incorporates SID,which is more sensitive to the spectrum as the measurement method and is more suitable for the frankincense spectral data compared with SVM.Results:For the origin classification task,samples of all levels from each origin were,respectively,selected for three-way classification.We used 10-fold cross-validation to select a model parameter in the experiment.When obtaining the optimal parameters,we randomly selected the training set and testing set,where the training set accounts for 70%and the training set for 30%.After repeating this random process 10 times,we obtained the final average classification accuracy,which is higher than 90%,and the standard deviation fluctuation is usually small.For the level classification task,samples of each level from three origins were separately selected for multiclassification.We randomly selected the training set and testing set from each origin.The level classification results of the three origins are good on D4350 data,and the classification accuracy of each level is basically above 80%.Conclusion:Experiments and analysis show that our algorithm framework has excellent classification performance,which is stable in origin classification and has potential for generalization.In addition,the experiments show that in our algorithm framework,different classification tasks need to combine different data sources to achieve better classification and recognition,as the origin classification task uses frankincense’s D3000 data,and level classification task uses frankincense’s D4350 data.