1.Comparison of Wild and Cultivated Codonopsis pilosula Based onTraditional Quality Evaluation
Xiaoyan LAN ; Chunfang TIAN ; Zhilai ZHAN ; Li ZHOU ; Xiang LI ; Zidong QIU ; Tiegui NAN ; Qili YUAN ; Xiaoliang LIN ; Congkui TIAN ; Meilan CHEN ; Liping KANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):156-164
ObjectiveTo conduct a systematic comparative study on wild and cultivated Codonopsis pilosula(CP) from three aspects, including characters, microscopy, and contents of primary and secondary metabolites. MethodWild and cultivated CP samples were collected, their characters were measured using vernier caliper, tape measure and balance, the paraffin sections were stained with safranin-fixed green dyeing, and their microstructure were observed under the optical microscope. The content of alcohol-soluble extracts in wild and cultivated CP was determined according to the method for determination of extract under CP in the 2020 edition of Chinese Pharmacopoeia, the starch content was determined by anthrone colorimetry, the content of total polysaccharides was determined by kit method, Fiber analyzer was used to determine the content of fiber components, and ultra performance liquid chromatography(UPLC) was used to determine the content of monosaccharides, disaccharides and some secondary metabolites. Multivariate statistical analysis methods such as principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to screen key differential components between wild and cultivated CP on the basis of variable importance in the projection(VIP) value>1 and P<0.05. ResultIn terms of morphological characteristics, the "lion's head-like" shape, longitudinal wrinkles, and circumferential wrinkles below the root cap of wild CP were more pronounced in wild CP compared to the cultivated ones. Regarding transverse sectional features, wild CP had more fissures on the outer side of the cortex and a larger duramen. Under microscopic examination, wild CP had more stone cells, a larger proportion of xylem, and the presence of cork cells arranged in rings in the xylem, while cultivated CP has a larger proportion of phloem, smaller vessel diameters, and a more loosely arranged vascular system. In terms of primary metabolites, the contents of 45% ethanol-soluble extract and total polysaccharides in cultivated CP were significantly higher than those in the wild ones(P<0.05), the contents of lignin, hemicellulose, cellulose, fructose and glucose in wild CP were significantly higher than those in the cultivated ones(P<0.05), while sucrose content in the cultivated CP was significantly higher than that in the wild ones(P<0.05). Concerning secondary metabolites, the contents of tryptophan and tangshenoside Ⅰ in cultivated CP were significantly higher than those in the wild ones(P<0.05), whereas the contents of lobetyolinin, lobetyol and atractylenolide Ⅲ in wild CP were significantly higher than those in the cultivated ones(P<0.05). ConclusionThere are significant differences between wild and cultivated CP in terms of morphological characteristics, microscopic features and chemical composition. Glucose, fructose, sucrose, tangshenoside Ⅰ, tryptophan and cellulose components are the key differential components between wild and cultivated CP. Wild CP contains more polyacetylenes and fructose, whereas cultivated CP has higher levels of tangshenoside Ⅰ and sucrose, with noticeably lower cellulose content. These distinctions may be related to their growth conditions, growth years and cultivation techniques. Based on the results of this study, it is recommended to increase polyacetylenes and the content ratio of fructose to sucrose as an indicators to characterize different production methods of CP, in order to guide the high-quality production of CP.
2.Effects of mild moxibustion on intestinal microbiome and NLRP3 inflammasome in rats with 5-fluorouracil-induced intestinal mucositis.
Bing-Rong LI ; Shi-Yun SHAO ; Long YUAN ; Ru JIA ; Jian SUN ; Qing JI ; Hua SUI ; Li-Hong ZHOU ; Yi ZHANG ; Hui LIU ; Qi LI ; Yan WANG ; Bi-Meng ZHANG
Journal of Integrative Medicine 2021;19(2):144-157
OBJECTIVE:
The present study investigated how mild moxibustion treatment affects the intestinal microbiome and expression of NLRP3-related immune factors in a rat model of intestinal mucositis (IM) induced with 5-fluorouracil (5-Fu).
METHODS:
Forty male Sprague-Dawley rats were randomly divided into control, chemotherapy, moxibustion and probiotics groups. The IM rat model was established by intraperitoneal injection of 5-Fu. Mild moxibustion treatment and intragastric probiotic administration were provided once daily for 15 days. Tissue morphology, serum levels of inflammatory factors and the expression levels of tight junction proteins, caspase-1, gasdermin D and NLRP3 were evaluated in colon tissue, through hematoxylin and eosin staining, electron microscopy, enzyme-linked immunosorbent assay, Western blotting, quantitative real-time reverse transcription polymerase chain reaction and immunofluorescence. Gut microbiome profiling was conducted through 16S rRNA amplicon sequencing.
RESULTS:
Moxibustion and probiotic treatments significantly increased the expression levels of tight junction proteins, reduced cell apoptosis and the expression levels of caspase-1, gasdermin D and NLRP3; they also decreased the serum levels of tumor necrosis factor-α, interleukin (IL)-6, IL-1β and IL-18, while increasing serum levels of IL-10. Moxibustion and probiotic treatments also corrected the reduction in α-diversity and β-diversity in IM rats, greatly increased the proportion of the dominant bacterial genus Lactobacillus and reduced the abundance of the genera Roseburia and Escherichia in chemotherapy-treated rats to levels observed in healthy animals. We also found that these dominant genera were firmly correlated with the regulation of pyroptosis-associated proteins and inflammatory factors. Finally, moxibustion and probiotic treatments elicited similar effects in regulating intestinal host-microbial homeostasis and the expression of NLRP3 inflammasome-related factors.
CONCLUSION
Moxibustion exerts its therapeutic effect on IM by ameliorating mucosal damage and reducing inflammation. Moreover, moxibustion modulates the gut microbiota, likely via decreasing the expression levels of the NLRP3 inflammasome.