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SARS-CoV-2 infection-induced hyper-inflammation links to the acute lung injury and COVID-19 severity. Identifying the primary mediators that initiate the uncontrolled hypercytokinemia is essential for treatments. Mast cells (MCs) are strategically located at the mucosa and beneficially or detrimentally regulate immune inflammations. Here we showed that SARS-CoV-2-triggeed MC degranulation initiated alveolar epithelial inflammation and lung injury. SARS-CoV-2 challenge induced MC degranulation in ACE-2 humanized mice and rhesus macaques, and a rapid MC degranulation could be recapitulated with Spike-RBD binding to ACE2 in cells; MC degranulation alterred various signaling pathways in alveolar epithelial cells, particularly, led to the production of pro-inflammatory factors and consequential disruption of tight junctions. Importantly, the administration of clinical MC stabilizers for blocking degranulation dampened SARS-CoV-2-induced production of pro-inflammatory factors and prevented lung injury. These findings uncover a novel mechanism for SARS-CoV-2 initiating lung inflammation, and suggest an off-label use of MC stabilizer as immunomodulators for COVID-19 treatments. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/449680v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@899996org.highwire.dtl.DTLVardef@1c26c0eorg.highwire.dtl.DTLVardef@1442cdcorg.highwire.dtl.DTLVardef@dd4204_HPS_FORMAT_FIGEXP M_FIG C_FIG In BriefSARS-CoV-2 triggers an immediate mast cell (MC) degranulation, which initiates the alveolar epithelial inflammation and disrupts the tight junction. MC stabilizers that block degranulation reduce virus-induced lung inflammation and injury. HighlightsO_LIThe binding of RBD of Spike protein of SARS-CoV-2-to ACE2 receptor protein triggers an immediate MC degranulation C_LIO_LIMC degranulation induces transcriptomic changes include an upregulated inflammatory signaling and a downregulated cell-junction signaling C_LIO_LIMC degranulation leads to alveolar epithelial inflammation and disruption of tight junctions C_LIO_LIMC stabilizer that inhibits degranulation reduces SARS-CoV-2-induced lung inflammation and injury in vivo C_LI
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Objective: To analyze and compare UPLC fingerprints of root, rhizome, stem, and leaf of Stephania tetrandra, learn the differences in chemical component types and contents of main active components, and provide basis for rational development and utilization of S. tetrandra. Methods: UPLC was used to obtain characteristic chromatograms of different parts; The Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (Version 2012) was run to capture the common peaks of different parts and calculate their similarity and analyze the characteristic peaks of different parts. SPSS 23.0 was run to compare the difference in component contents of the roots and rhizomes using the paired sample t-test. Results: The similarity in chemical composition between root and rhizome was 0.928, indicating they have similar chemical composition, and both of them contained tetrandrine and fangchinoline, the index components. The similarity between rhizome and leaf was 0.947; The similarity was low between stem, leaf and root and rhizome, and there were no tetrandrine and fangchinoline in the first two parts. The results of paired samples t-test show that the total content of chemical components in rhizome was higher than that in roots, and the mainly difference came from other non-index components, but there was no significant difference between tetrandrine and fangchinoline. Conclusion: Significant differences are present in chemical composition types and contents of different medicinal parts of S. tetrandra; The type of chemical components in rhizome is similar to that in root, and the content of some components in rhizome is significantly higher than that in root, which means that rhizomes can be used as an equivalent of roots. Stems and leaves cannot be a substitute for roots because they do not contain tetrandrine and fangchinoline, but they contain many other chemical components which can be utilized as a new resource.
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Objective:To investigate the quality regionalization and environmental impact factors of Stephaniae Tetrandrae Radix based on main active ingredients，and provide a reference for the determination of high-quality production areas and the dominant environmental factors affecting the content of Stephaniae Tetrandrae Radix. Method:Partial least squares regression analysis (PLS) and principal component analysis (PCA) were used to study the quality regionalization and environmental impact factors based on the main active ingredients of tetrandrine and fangchinoline, and investigate the environmental factors of the producing areas. Result:The content of fangchinoline was positively correlated with soil pH and annual average temperature，negatively correlated with latitude. The content of tetrandrine was positively correlated with soil pH，negatively correlated with annual rainfall and longitude. The total content was positively correlated with soil pH and annual average temperature，but negatively correlated with annual rainfall，latitude and longitude. Principal component analysis showed that the 50 production areas could be divided into four groups of quality formation. The groups with the highest scores were Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan，all of which were best areas for accumulation of the two main active ingredients. Conclusion:Soil pH，annual average temperature，annual rainfall，latitude and longitude are the main environmental factors affecting the main active ingredients of Stephaniae Tetrandrae Radix. The best areas for accumulation of tetrandrine and fangchinoline are Shixing county in Guangdong，Shexian county in Anhui，Songxi county in Fujian，Nanxiong city in Guangdong and Xiangxiang city in Hunan.
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Objective::Sixty-nine germplasm samples of Picria felterrae collected from the main producing areas in Guangxi were subject to genetic diversity and genetic relationship analyses using the simple seguence repeat(SSR) molecular marker technology and good germplasm genes associated with the content of picfeltarraenins were screened so as to provide references for germplasm resource evaluation, phylogenetic analysis, and molecular mark assisted breeding of that species. Method::20 pairs of randomly selected primers were amplified based on the transcriptome sequencing technology. The genetic diversity of and genetic relationship between the 69 samples were analyzed using the genetic polymorphic information for each marker locus, and one-variable linear regression and multiple stepwise regression analyses were performed to screen molecular markers associated with the content of picfeltarraenins. Result::The amplification using the 20 pairs of SSR primers produced 76 alleles, 3.8 alleles for each locus on average, higher than effective alleles (1.969 2), and the rare allele rate was 38.2%, suggesting that the alleles distributed unequally. The polymorphism rates of alleles varied between 0-59%, with an average of 38.24%, showing a great difference among loci. The polymorphic information content (PIC) varied between 0-0.621 1, with an average of 0.378 0.Shannon polymorphic information index varied between 0-1.240 1, with an average of 0.759.Nei's gene diversity index (Nei) varied between 0-0.682 3, with an average of 0.440 9.P21 had the highest level accompanied with the lowest P7 for the above three indexes, and significant genetic diversity differences were identified among the loci. For all loci, the mean observed heterozygosity was 0.382 4, lower than the average expected heterozygosity of 0.442 5, suggesting the loss of heterozygosity, the average genetic differentiation coefficient (Fst) was 0.365 9 and the average gene flow Nm was 0.433 2, suggesting a high genetic divergence and a low gene flow. The results of one-variable linear regression and multiple stepwise regression showed that there were 5 loci related to each of the picfeltarraenin IA and IB, and only 1 loci was associated with the content of both. Conclusion::There were significant differences in the genetic diversity of 20 SSR marker sites, and the 69 germplasm samples were greatly genetically differentiated and had low gene flow. From the selected 20 SSR markers 9 marker loci associated with the content of picfeltarraenin IA and IB were selected. The results can be used as a reference for phylogenetic analysis and selective breeding of Picria felterrae.
