1.Effects of Processed Temperature And Time on Color and Contents of Six Types of Boswellic Acids in Frankinense
Zhangchi NING ; Zhiqian SONG ; Chun WANG ; Yuanyan LIU ; Honglian ZENG ; Jiahe GAN ; Xinling MA ; Zhenli LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(3):508-515
This paper focused on factors which affected on different color of northern and southern region vinegar-processed frankincense.Meanwhile,contents of six main boswellic acids were also determined to elaborate the influence of heat in chemical components.Vinegar-processed frankincense from northern and southern region was collected.And different temperature and time were used in the processing of frankincense to receive the vinegar-processed frankincense samples.The color difference meter was utilized combining with the PCA statistic analysis method.The Zorbax ExtendC18 chromatographic column (4.6 mm × 50 mm,1.8 μm) was used with acetonitrile-0.1% phosphoric acid as the mobile phase and gradient elution.The velocity of flow was 1 mL· min-1.The detection wavelength was 210 nm and 250 nm.The column temperature was 30℃.The results showed that the color of northern region processed frankincense was yellow or pale brown.And the southern region processed frankincense was pale brown or dark brown.It showed the difference on processed degree.The L* value of the northern processed frankincense was 75.327 to 80.746 and the L* value of southern processed was 44.321 to 49.527.The a* value of the northern processed frankincense was 5.378 to 6.502 and the a* value of southern processed was 9.423 to 9.978.There was no significant difference on b*.There were certain differences on L* and a* among vinegar-processed frankincense with the same surface color.The color parameter results of self-made vinegar-processed frankincense indicated that along with changes of processing temperature and time,the color,L* and a* change.Even frankincense processed for 30 min with mild fire,it will not achieve the color parameter value of the southern region vinegar-processed frankincense.However,after 11 min processing with medium fire,the color can be achieved.The content determination results showed that four contents,including α-boswellic acid,β-boswellic acid,3-acetyl-α-boswellic acid and 3-acetyl-β-boswellic acid were increased.Contents of 11-carbonyl-3-boswellic acid and 3-acetyl-11-carbonyl-β-boswellic were decreased after being processed.The range of increasing or decreasing by medium fire was higher than mild fire.At the same temperature,as the increasing of processing time,the content has an increasing or decreasing tendency.It was concluded that temperature was the main factor influencing the color of vinegar-processed frankincense from northern and southern regions.Different processing degrees also make influence on the contents of chemical compounds.The color parameter value can be used to evaluate the color of processed frankincense.
2.Exploring Synergistic Mechanism of Vinegar-processed Olibanum in Treatment of Ulcerative Colitis Based on Bile Acid-regulated "Gut-liver" Crosstalk
Shitao PENG ; Zhenli LIU ; Zhiqian SONG ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(21):59-68
ObjectiveTo investigate the synergistic mechanism of vinegar-processed Olibanum on ulcerative colitis(UC) via the bile acids regulating "gut-liver" crosstalk. MethodRats were randomly divided into normal group, model group, Olibanum group and vinegar-processed Olibanum group. UC model of rats was induced by intracolonic instillation of 2,4,6-trinitrobenzenesulfonic acid(TNBS). Ultra high performance liquid chromatography-triple quadrupole-mass spectrometry(UPLC-QQQ-MS) was used to perform the qualitative analysis of 30 bile acids in the colon of rats. Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to detect changes in the expression of farnesoid X receptor(FXR), fibroblast growth factor 15(FGF15) and FGF receptor 4(FGFR4) in "gut-liver" crosstalk at mRNA and protein levels. And with the help of HcoEpiC cell model intervened by conjugated bile acids, simulating the UC state, and according to the different modes of intervention, they were divided into the blank group, conjugated bile acid group, Olibanum group, vinegar-processed Olibanum group and 3-O-acetyl-9,11-dehydro-β-boswellic acid(ADHBA) group. The effect of Olibanum before and after processing with vinegar and the main differential component ADHBA on the mRNA expression of FXR and FGF19 were explored by Real-time PCR. ResultCompared with the normal group, the levels of conjugated bile acids in the model group increased significantly(P<0.01), and the mRNA and protein expressions of "gut-liver" crosstalk factors FXR, FGF15 and FGFR4 decreased significantly(P<0.05, P<0.01). Compared with the model group, the content of conjugated bile acids in the Olibanum group and vinegar-processed Olibanum group was significantly decreased(P<0.01), the mRNA and protein expressions of FXR, FGF15 and FGFR4 were significantly elevated(P<0.05, P<0.01), and vinegar-processed Olibanum exhibited superior effects than Olibanum. In cellular experiments, a significant decrease in mRNA expression of FXR and FGF19 was observed in the conjugated bile acid group when compared with the blank group(P<0.01). Compared with the conjugated bile acid group, the mRNA expressions of FXR and FGF19 were significantly higher in the Olibanum, vinegar-processed Olibanum and ADHBA groups(P<0.05, P<0.01), and the effect of vinegar-processed Olibanum was more favorable. ConclusionVinegar-processed Olibanum may enhance the ameliorating effect on UC by enhancing the down-regulation of conjugated bile acids in the colon and the up-regulation of FXR-FGF15/19-FGFR4 "gut-liver" crosstalk pathway, and ADHBA may be the main material basis for the synergy.
3.Exploring Synergistic Mechanism of Processing with Vinegar on Olibanum in Treatment of Ulcerative Colitis Based on Intestinal Flora
Wenjie LU ; Shitao PENG ; Zhiqian SONG ; Zhenli LIU ; Chun WANG ; Zhangchi NING
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):182-191
ObjectiveTo explore the improving effects and its synergistic mechanism of Olibanum before and after processing with vinegar on glycodesoxycholic acid(GDCA) intervention in mice with ulcerative colitis(UC) based on the perspective of intestinal flora. MethodC57BL/6J male mice were randomly divided into the normal group, model group, GDCA group, Olibanum group(1.5 g·kg-1) and vinegar-processed Olibanum(1.5 g·kg-1) group, with 6 mice in each group. Mice in the normal group drank water freely, and mice in the other groups were given 2% dextran sulfate sodium(DSS) periodically to establish a UC mouse model. During the modeling, GDCA group, Olibanum group and vinegar-processed Olibanum group were intervened by intraperitoneally injection of GDCA(0.05 g·kg-1). From the 13th day after modeling, Olibanum group and vinegar-processed Olibanum group were given the corresponding doses of drugs by gavage, once a day, for 36 d. During this period, the body mass of mice was recorded and the disease activity index(DAI) was assessed. On day 48, faeces were collected for 16S rRNA and metagenomic sequencing to analyse changes in intestinal flora. On the 49th day, hematoxylin-eosion(HE) staining was used to observe the colon histological lesions, enzyme-linked immunosorbent assay(ELISA) was used to determine serum levels of tumour necrosis factor-α(TNF-α), interleukin(IL)-1β and IL-6, and Spearman correlation analysis was used to explore the correlation between differential bacterial species and inflammatory factor levels. ResultCompared with the normal group, the model group showed a significant decrease in body weight(P<0.01), a significant increase in DAI(P<0.05), and a significant increase in TNF-α, IL-1β and IL-6 levels(P<0.01), and there was partial infiltration of inflammatory cells in the colon. Compared with the model group, mice in the GDCA group showed a significant decrease in body weight, a significant increase in DAI and levels of TNF-α, IL-1β and IL-6(P<0.01), and severe disruption of colonic crypt structure, extensive infiltration of inflammatory cells, and a significant decrease in goblet cells. Compared with the GDCA group, both the Olibanum and vinegar-processed Olibanum groups showed a significant recovery in body weight, a significant decrease in DAI and levels of TNF-α, IL-6 and IL-1β(P<0.05, P<0.01), and the modulating effect of vinegar-processed Olibanum was significantly better than that of Olibanum. Alpha diversity showed that Chao1 index of UC mice significantly increased(P<0.01) and Shannon index decreased significantly(P<0.05) in UC mice after GDCA intervention. Beta diversity showed that the microbial community structure of the 5 groups had significant changes, Olibanum and vinegar-processed Olibanum could modulate the changes in the structure of the intestinal flora in UC mice after GDCA intervention. Microbial sequencing results indicated that, compared with the normal group, the Firmicutes/Bacteroidetes ratio in the model group was significantly higher(P<0.05), and the relative abundance of 3 genera and 5 species of flora changed significantly(P<0.05, P<0.01). Compared with the model group, the Firmicutes/Bacteroidetes ratio in the GDCA group was significantly higher(P<0.05), the relative abundance of 7 pathogenic bacterial genera and four species was significantly increased(P<0.05, P<0.01), and the relative abundance of three beneficial bacterial genera and Bacteroides_intestinalis was significantly decreased(P<0.05, P<0.01). Olibanum group and vinegar-processed Olibanum group could modulate the Firmicutes/Bacteroidetes ratio, the relative abundance of pathogenic bacteria and beneficial bacteria, and the vinegar-processed Olibanum group was significantly superior to Olibanum group in terms of modulating the Firmicutes/Bacteroidetes ratio, the relative abundance of the three genera and five species of bacteria(P<0.01, P<0.05). Correlation analysis showed that the relative abundance of Bacteroides_intestinalis was negatively correlated with the levels of TNF-α, IL-6 and IL-1β, the relative abundance of Prevotella_sp_CAG873, Bacteroides_sp_CAG927, Bacteroidales_bacterium_52_46 and Bacteroidales_bacterium was positively correlated with TNF-α, IL-6 and IL-1β levels. ConclusionGDCA can exacerbate UC colonic inflammation, and Olibanum and vinegar-processed Olibanum have an ameliorative effect on GDCA-mediated UC, with the vinegar-processed Olibanum showing a stronger ameliorative effect, the mechanism may be related to the regulation the abundance and structure of intestinal beneficial and pathogenic bacteria, and the reduction of inflammatory factor levels.