Dicliptera chinensis (L.) Juss. is a herbaceous plant renowned for its anti-inflammatory and antioxidant properties. Previous studies have demonstrated that its polysaccharide (DCP) exerts hepatoprotective effects, yet the underlying mechanism by which DCP alleviates metabolic dysfunction-associated steatotic liver disease (MASLD) remains unclear. This study investigated the hepatoprotective effects of DCP in high-glucose and high-fat (HHF) diet-induced MASLD mice and AML12 hepatocytes, with a focus on miRNA-mediated regulatory mechanisms. Small RNA sequencing revealed that miR-3073b-5p was significantly upregulated in MASLD. Dual-luciferase reporter assays verified the direct binding of miR-3073b-5p to the 3'UTR of CAMKK2, and RIP assays further confirmed their interaction under physiological conditions. In vivo, DCP administration significantly ameliorated hyperglycemia, dyslipidemia, hepatic steatosis, and oxidative injury. 16S rRNA sequencing and bile acid metabolomics analyses demonstrated that DCP effectively reshaped the gut microbiota composition and restored bile acid metabolic homeostasis. In vitro, DCP downregulated miR-3073b-5p expression, thereby relieving the suppression of CAMKK2, regulating the AMPK/mTOR/Nrf2 signaling axis, restoring autophagy, and counteracting ferroptosis. These findings indicate that DCP alleviates MASLD by regulating miR-3073b-5p/CAMKK2 via the gut microbiota-bile acid axis, positioning it as a promising natural polysaccharide for MASLD therapy and providing a novel molecular target for the targeted intervention of this disease.

ETHNOPHARMACOLOGICAL RELEVANCE:Scoparone (SCO) is a bioactive coumarin compound isolated from the traditional medicinal plant Artemisia dracunculus L. var. turkestanica Krasch. Previous studies have demonstrated that SCO ameliorates hepatic steatosis. However, the precise molecular mechanisms underlying its action, especially in metabolic dysfunction-associated steatotic liver disease (MASLD), remain incompletely elucidated. AIM OF THE STUDY:The study aimed to reveal the therapeutic effects and underlying mechanisms of SCO in treating MASLD. MATERIALS AND METHODS:In this study, an in vivo MASLD model was constructed in mice through high-fat diet (HFD) induction, and an in vitro steatosis model was established by treating AML-12 hepatocytes with sodium oleate (NaOA). Hepatic histopathological characteristics, lipid deposition and biochemical parameters of all experimental groups were detected, differentially expressed genes and miRNAs were screened through RNA and miRNA sequencing, and the direct targeting effect of miR-3073a-3p on Camkk2 was verified using dual-luciferase reporter gene assay. Meanwhile, Western blot and qRT-PCR were applied to determine cellular functional indicators as well as the protein and mRNA expression levels of key signaling molecules. RESULTS:miR-3073a-3p directly bound to the 3'UTR of Camkk2 mRNA, suppressing CAMKK2 expression and inhibiting CAMKK2/AMPK signaling. Overexpression of miR-3073a-3p exacerbated hepatic steatosis, mitochondrial dysfunction, oxidative stress, bile acid imbalance and autophagy impairment, whereas miR-3073a-3p knockdown ameliorated MASLD phenotypes in a CAMKK2-dependent manner. SCO treatment downregulated miR-3073a-3p expression, restored CAMKK2 levels and activated protective pathways mediated by Nrf2, FXR and ULK1, thereby enhancing antioxidant defense, restoring bile acid homeostasis, promoting autophagy and alleviating liver injury. CONCLUSION:By modifying the miR-3073a-3p/CAMKK2 axis, SCO improved MASLD, indicating its potential therapeutic utility in the management of metabolic liver disorders.

Curcuma kwangsiensis S.G. Lee et C.F. Liang, a traditional medicinal plant, has rhizomes (RH) and tuberous roots (TR) listed in the Chinese Pharmacopoeia for distinct therapeutic applications, while fibrous roots (FR) are often discarded. Terpenoids, C. kwangsiensis's main active components, show unclear differential accumulation patterns among underground tissues with different morphologies. This study aimed to reveal terpenoid differential accumulation patterns and molecular regulatory mechanisms in RH, TR, and FR. A comprehensive analysis combining full-spectrum non-targeted metabolomics (UPLC-MS/MS and GC-MS) with transcriptomics (Iso-seq and RNA-seq) was used to establish the terpenoid metabolism-gene regulatory network. Total terpenoid content in C. kwangsiensis was highest in FR > RH > TR. UPLC-MS/MS and GC-MS identified 39, 41, and 34 differentially accumulated metabolites (DAMs) in RH versus TR, RH versus FR, and TR versus FR, respectively. Upregulated monoterpenoids and sesquiterpenoids DAMs were higher in RH and FR than TR. Transcriptomics identified 78 differentially expressed genes (DEGs). qRT-PCR results validated the RNA-seq findings, showing consistent expression profiles for TPS genes, as well as upstream and downstream genes in the terpenoid backbone biosynthesis. Significant DEGs and the accumulation of DAMs correlations were identified through Pearson correlation analysis. Further Spearman analysis uncovered significant regulatory relationships between 42 WRKY transcription factors and 5 TPS genes. The terpenoid metabolism-gene regulatory network in C. kwangsiensis was established, elucidating the mechanism of the differential terpenoid accumulation in RH, TR, and FR. These results offer scientific support for improving the quality of C. kwangsiensis's underground tissues and the high-value utilization of non-traditional medicinal parts.

Background Acute liver failure (ALF) is associated with ferroptosis and currently lacks specific drugs and therapies. Previous studies have shown the hepatoprotective activity of Sophoricoside (SOP) in diverse models of hepatic injury. However, the role of SOP in ALF caused by lipopolysaccharide (LPS)/D-galactosamine (D-GalN) is still unknown. Purpose To investigate the therapeutic effects of SOP on ALF and its underlying mechanisms of action. Methods In vivo, an ALF mouse model was established using LPS and D-GalN. RNA sequencing was used to identify differentially expressed genes, which were subsequently confirmed by Western blot and qRT-PCR. In vitro, erastin was used to induce ferroptosis in HepG2 cells. Subsequently, the cells were treated with SOP, the nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor ML385, and the glutathione peroxidase 4 (GPX4) inhibitor RSL3. Additionally, siRNA was utilized for Nrf2 knockdown. Measurements of biochemical markers, reactive oxygen species (ROS) levels, alterations in mitochondrial membrane potential, qRT-PCR, and Western blot analysis were used to confirm the effectiveness and mechanism of SOP. Finally, the interaction between SOP and Nrf2 was verified through molecular docking, surface plasmon resonance (SPR), and cellular thermal shift assay (CETSA). Results In vivo, experimental analysis revealed that SOP ameliorated liver injury, hepatocyte Fe2+ accumulation, and oxidative stress. RNA sequencing analysis indicated that SOP reversed the abnormal expression of genes related to Nrf2 and the ferroptosis pathway. In vitro, SOP ameliorated erastin-induced oxidative stress, ROS accumulation, and mitochondrial membrane potential loss. After the application of pharmacological inhibitors and siRNA knockdown, the SOP’s inhibitory effect on ferroptosis was antagonized. Conclusion SOP alleviated ALF by activating the Nrf2/GPX4 signaling pathway, thereby mitigating oxidative stress and ferroptosis.

To elucidate the mechanism by which steaming affects the quality of Curcuma kwangsiensis root tubers, methods such as LSCM, RVA, dual-wavelength spectrophotometry, LF-NMR, and LC-MS were employed to qualitatively and quantitatively detect changes in starch gelatinization characteristics, water distribution, and material composition of C. kwangsiensis root tubers under different steaming durations. Based on multivariate statistical analysis, the correlation between differences in gelatinization parameters, water distribution, and terpenoid material composition was investigated. The results indicate that steaming affects both starch gelatinization and water distribution in C. kwangsiensis. During the steaming process, transformations occur between amylose and amylopectin, as well as between semi-bound water and free water. After 60 min of steaming, starch gelatinization and water distribution reached an equilibrium state. The content of amylopectin, the amylose-to-amylopectin ratio, and parameters such as gelatinization temperature, viscosity, breakdown value, and setback value were significantly correlated(P≤0.05). Additionally, the amylose-to-amylopectin ratio was significantly correlated with total free water and total water content(P≤0.05). Steaming induced differences in the material composition of C. kwangsiensis root tubers. Clustering of primary metabolites in the OPLS-DA model was distinct, while secondary metabolites were classified into 9 clusters using the K-means clustering algorithm. Differential terpenoid metabolites such as(-)-α-curcumene were significantly correlated with zerumbone, retinal, and all-trans-retinoic acid(P<0.05). Curcumenol was significantly correlated with isoalantolactone and ursolic acid(P<0.05), while all-trans-retinoic acid was significantly correlated with both zerumbone and retinal(P<0.05). Alpha-tocotrienol exhibited a significant correlation with retinal and all-trans-retinoic acid(P<0.05). Amylose was extremely significantly correlated with(-)-α-curcumene, curcumenol, zerumbone, retinal, all-trans-retinoic acid, and α-tocotrienol(P<0.05). Amylopectin was significantly correlated with zerumbone(P<0.05) and extremely significantly correlated with(-)-α-curcumene, curcumenol, zerumbone, retinal, all-trans-retinoic acid, and 9-cis-retinoic acid(P<0.01). The results provide scientific evidence for elucidating the mechanism of quality formation of steamed C. kwangsiensis root tubers as a medicinal material.

Ethnopharmacological relevance Taraxasterol (TAR), a compound highly abundant and easily obtainable from Tibetan medicine Ixeridium gramineum (Fisch.) Tzvel., exhibits a variety of biological effects, including hepatoprotective, anti-inflammatory, and antioxidant activities.Aim of the study: To investigated the protective role and underlying mechanisms of TAR in fulminant hepatitis (FH) through the regulation of oxidative stress, inflammatory responses, and apoptosis by modulating the JAK/STAT and TNF signalling pathways. Material and methods The study used Kunming mice to establish a D-GalN/LPS-induced FH model, which was divided into the following groups: Control group, D-GalN/LPS group, D-GalN/LPS + Silymarin group, D-GalN/LPS + TAR 2.5 group, D-GalN/LPS + TAR 5 group, D-GalN/LPS + TAR 10 group, and TAR 10 group. H&E staining and biochemical analyses were employed to evaluate liver pathological changes. Oxidative stress factors and inflammatory response were assessed via ELISA. RNA sequencing analysis was used to detect changes in inflammatory factor genes and apoptosis genes with TAR intervention in liver tissues. The distribution of the proteins p-STAT3 and p-JNK in liver tissues was ascertained using immunohistochemical staining. In vitro experiments were conducted on RAW264.7 cells exposed to LPS and TAR. Apoptosis was evaluated via flow cytometry and Hoechst 33258 staining. Immunofluorescence staining was employed to determine the protein expression levels of p-STAT3 and p-JNK in RAW264.7 cells. Gene and protein expression in the JAK/STAT and TNF signalling pathways, as well as apoptosis, were analyzed using qRT-PCR and Western blotting. Results TAR effectively reduced hepatocyte necrosis, diminished inflammatory factor release, inhibited oxidative stress, significantly decreased the apoptosis of RAW264.7 cells, inhibited the protein expressions of p-JAK2, p-STAT3, p-MEK4, p-JNK, Caspase-3, Caspase-8, and Bax, and increased the protein expressions of SOCS3 and Bcl-2. Conclusion TAR prevents D-GalN/LPS-induced FH by regulating the JAK/STAT and TNF signalling pathways and apoptosis, demonstrating its therapeutic potential in treating liver diseases.

BACKGROUND:Isoniazid and rifampicin, frontline tuberculosis drugs, frequently induce drug-induced liver injury (DILI), marked by hepatitis and hepatocyte necrosis. Polysaccharides from Dicliptera chinensis (L.) Juss. (DCP) exhibit anti-inflammatory, antioxidant, and hepatoprotective properties, but their effects on DILI remain unexplored OBJECTIVE: This study investigated DCP therapeutic potential against DILI and elucidated its molecular mechanisms METHODS: In vivo (using C57BL/6 mice) and in vitro (using HepG2 cells) DILI models were established and treated with DCP. Transcriptomics, qRT-PCR, and Western blotting were employed to analyze pathway regulation RESULTS: DCP significantly attenuated hepatocyte apoptosis, inflammation, and oxidative stress in DILI mice. Transcriptomic analysis linked DCP'S effects to the modulation of AMPK-FOXO3, p53, and NF-κB pathways, alongside regulation of antioxidant and cell cycle genes. In HepG2 cells, DCP similarly protected against DILI by enhancing AMPK phosphorylation, which facilitated the FOXO3 nuclear translocation. Both models demonstrated DCP'S suppression of p53 and NF-κB activation, restoration of antioxidant defenses, and correction of cell cycle dysregulation CONCLUSION: DCP mitigates DILI by reducing apoptosis, oxidative stress, and inflammation through activation of the AMPK-FOXO3 pathway, inhibition of p53/NF-κB signaling, and stabilization of the cell cycle. These findings highlight DCP'S potential as a therapeutic agent for DILI prevention and treatment.

ETHNOPHARMACOLOGICAL RELEVANCE:Polygala fallax Hemsl. is a traditional folk medicine commonly used by ethnic minorities in the Guangxi Zhuang Autonomous Region, and has a traditional application in the treatment of liver disease. Polygala fallax Hemsl. polysaccharides (PFPs) are of interest for their potential health benefits. AIM OF THIS STUDY:This study explored the impact of PFPs on a mouse model of cholestatic liver injury (CLI) induced by alpha-naphthyl isothiocyanate (ANIT), as well as the potential mechanisms. MATERIALS AND METHODS:A mouse CLI model was constructed using ANIT (80 mg/kg) and intervened with different doses of PFPs or ursodeoxycholic acid. Their serum biochemical indices, hepatic oxidative stress indices, and hepatic pathological characteristics were investigated. Then RNA sequencing was performed on liver tissues to identify differentially expressed genes and signaling pathways and to elucidate the mechanism of liver protection by PFPs. Finally, Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting were used to verify the differentially expressed genes. RESULTS:Data analyses showed that PFPs reduced the levels of liver function-related biochemical indices, such as ALT, AST, AKP, TBA, DBIL, and TBIL. PFPs up-regulated the activities of SOD and GSH, down-regulated the contents of MDA, inhibited the release of IL-1β, IL-6, and TNF-α, or promoted IL-10. Pathologic characterization of the liver revealed that PFPs reduced hepatocyte apoptosis or necrosis. The RNA sequencing indicated that the genes with differential expression were primarily enriched for the biosynthesis of primary bile acids, secretion or transportation of bile, the reactive oxygen species in chemical carcinogenesis, and the NF-kappa B signaling pathway. In addition, the results of qRT-PCR and Western blotting analysis were consistent with those of RNA sequencing analysis. CONCLUSIONS:In summary, this study showed that PFPs improved intrahepatic cholestasis and alleviated liver damage through the modulation of primary bile acid production, Control of protein expression related to bile secretion or transportation, decrease in inflammatory reactions, and inhibition of oxidative pressure. As a result, PFPs might offer a hopeful ethnic dietary approach for managing intrahepatic cholestasis.

Alcoholic liver disease (ALD) is a prevalent liver condition that arises from prolonged and excessive alcohol intake. Bergenin (BER) is an effective phytotherapeutic agent that exhibits pharmacological properties, including anti-inflammatory and anti-oxidative effects. To establish an in vivo model of ALD, C57BL/6 mice were continuously fed a high-fat diet (HFD) and administered alcohol gavage for 8 weeks, while concurrently administering BER and evaluated for therapeutic effects. After modeling, the therapeutic effects of BER were evaluated by observing histopathological changes and the detection of relevant biochemical indicators in mice. In addition, RNA sequencing of liver tissues was performed to analyze differentially expressed genes and to investigate the associated signaling pathways in order to elucidate the protective mechanisms of BER. These differentially expressed genes were mainly enriched in lipid metabolism pathways and the cytochrome P450 metabolism of exogenous substances. Subsequently, HepG2 was co-treated with sodium oleate (NaOA) and ethanol to establish an in vitro model, and the specific mechanism by which BER ameliorates ALD was further analyzed in depth. AMPK inhibitor, Compound C (CC), was demonstrated to significantly inhibit the regulation of lipid metabolism by BER in vitro. Finally, the differentially expressed genes selected were validated through qRT-PCR and Western blot analysis. Collectively, our findings revealed that BER effectively alleviated liver injury caused by alcohol and HFD in mice, significantly suppressing lipid deposition in ALD, enhancing alcohol metabolism, and mitigating oxidative stress.

ETHNOPHARMACOLOGICAL RELEVANCE:Cholestatic liver disease (CLD) is a disease characterized by cholestasis. Farnesoid X receptor (FXR) is a nuclear receptor that maintains homeostasis in bile acid metabolism. Studies have shown that gut microbiota interfered with the FXR pathway. Modulation of FXR to inhibit cholestasis has become a key measure in the treatment of CLD. In traditional folk medicine, Desmodium styracifolium (Osb.) Merr. was used as a primary treatment for gallstones, gonorrhea, jaundice, cholecystitis and other diseases. Modern pharmacological studies had also found that the herb has anti-calculus, anti-inflammatory, antioxidant, diuretic and liver damage. Therefore, we speculated that Desmodium styracifolium (Osb.) Merr. extracts (DME) could alleviate CLD through the FXR pathway and might be associated with the gut microbiota. However, studies of DME alleviating CLD through the FXR pathway have not been reported. AIM OF STUDY:To study the effect and mechanism of DME in relieving CLD through in vivo and in vitro experiments. MATERIALS AND METHODS:First, mice were administrated with alpha-naphthyl isothiocyanate (ANIT) to establish a CLD model in vivo. Meanwhile, HepG2 cells were induced by lithocholic acid (LCA) to establish the CLD model in vitro. To evaluate the therapeutic effect of DME on CLD mice, hematoxylin-eosin (HE) staining, and biochemical indicators were performed. The prototype of the blood components in mice serum was detected by ultra-high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). 16S rDNA sequencing was used to analyze the gut microbiota. Finally, the protein and mRNA expression of the FXR pathway in mice liver tissues or HepG2 cells were detected by Western blot, qRT-PCR, or immunofluorescence. RESULTS:Pathological testing and biochemical indexes showed that DME significantly reduced serum ALT, AST, ALP, TBIL, DBIL, TBA and liver TBA levels, and attenuated liver tissue injury, necrosis and jaundice in CLD mice. In addition, MetagenomeSeq analysis of gut microbiota showed that DME significantly up-regulated the abundance of Parvibacter, down-regulated the abundance of Paenalcaligenes, and regulated bile acid homeostasis. In terms of mRNA expression, DME significantly upregulated the mRNA levels of Nr1h4, Abcb11, Cyp7a1 and Slc10a1. Meanwhile, in terms of protein expression, DME significantly up-regulated the protein expression levels of FXR, BSEP, CYP7A1 and NTCP, which regulated bile acid homeostasis. Finally, the molecular docking results showed that the components of DME, such as Lumichrome, Daidzein and Folic acid, all had good binding ability with FXR, and the surface plasmon resonance (SPR) results also showed that both Lumichrome and Daidzein had a relatively high affinity with FXR. CONCLUSION:DME alleviated CLD through the FXR pathway, and the mechanisms might be associated with the gut microbiota.

Dicliptera chinensis (L.) Juss., is an herb known for its anti-inflammatory and anti-oxidant properties. In the previous studies, the chemical composition of the polysaccharide from Dicliptera chinensis (L.) Juss. (DCP) has been characterized as consisting of DCP1 and DCP2, of which DCP2 has hepatoprotective effects. The study examined the hepatoprotective potential of DCP2 against alpha-naphthyl isothiocyanate (ANIT)-induced cholestatic liver disease (CLD). In this study, RNA sequencing identified key research pathways involving bile acid metabolism, oxidative stress, and inflammation. Furthermore, qRT-PCR and Western blot analyses were conducted to further characterize these pathways. Additionally, the study included in vitro experiments with HepG2 cells to further investigate the effects of DCP2 on bile acid metabolism. In summary, the protective effect of DCP2 on the liver was reflected in alleviating the inflammatory response and oxidative stress, regulating the metabolism of bile acids, and mitigating liver damage caused by bile acids. This study further elucidated the hepatoprotective effects of DCP2 by examining its ability to counteract ANIT-induced CLD, suggesting that DCP2 is a promising biomacromolecule for hepatoprotection.

Alcoholic fatty liver disease (AFLD) is characterized by excessive lipid accumulation in the liver. This study aimed to investigate the protective effects and mechanisms of Polygala fallax Hemsl polysaccharides (PFPs) on AFLD. PFPs were purified and structurally characterized. An AFLD model was established in mice using alcohol and a high-fat diet. A significant reduction in hepatic steatosis was observed following PFPs treatment, evidenced by decreased fat deposition in liver tissues. Additionally, PFPs reduced various liver injury markers, increased levels of antioxidant enzymes, and improved significantly liver function. RNA sequencing revealed that PFPs improved lipid and CYP450 metabolic pathway abnormalities in AFLD mice. Furthermore, PFPs activated the AMPK pathway, reducing lipid accumulation and enhancing lipid metabolism. A HepG2 cell model treated with ethanol and oleic acid showed significant biochemical improvements with PFPs pretreatment, including reduced lipid accumulation and lower reactive oxygen species (ROS) levels. To further elucidate the AMPK and PFPs correlation in AFLD, an AMPK inhibitor (compound C) was used. In vitro and in vivo qRT-PCR and Western blot results confirmed that PFPs protected against AFLD by activating AMPK phosphorylation, regulating lipid synthesis, and inhibiting lipid accumulation. PFPs also modulated CYP2E1 and oxidative stress-related gene expression, affecting liver metabolism.

BACKGROUND:Alcoholic liver disease (ALD) is a severe public health concern worldwide and there is still a lack of effective treatments. Qiwei Jinggan Ling (QJL) has protective effects against various liver injuries, but its pharmacological action on ALD has received little attention. PURPOSE:To investigate the effect and mechanism of QJL on ALD in vivo and in vitro. METHODS:In vivo, an ALD mouse model was established by alcohol combined with a high-fat diet (HFD) and treated with QJL. Biochemical indicators, HE staining, and Oil Red O staining were employed to assess hepatic oxidative stress, steatosis, and alcohol metabolism. RNA sequencing analysis was performed, and the results were verified by qRT-PCR and Western blot to elucidate the hepatoprotective mechanism of QJL. In vitro, HepG2 cells were co-stimulated with NaOA (sodium oleate) and EtOH (ethanol), followed by intervention with Compound C (CC, AMPK inhibitor) and QJL-containing serum. Oil Red O, BODIPY (boron-dipyrromethene), and ROS (reactive oxygen species) staining were applied to validate the efficacy and mechanism of QJL-containing serum. The expression of AMP-activated protein kinase (AMPK) pathway-related factors was analyzed through qRT-PCR and Western blot for additional corroboration. Moreover, the key pharmacodynamic components of QJL were identified by UPLC-MS/MS and molecular docking. RESULTS:In vivo, QJL ameliorated liver structural disorders, steatosis, oxidative stress, and impaired alcohol metabolism, as indicated by biochemical indicators and histopathological assays. RNA sequencing analysis revealed that QJL reversed the expression of genes related to alcohol metabolism, fatty acid metabolism, and cholesterol metabolism. The results of qRT-PCR and Western blot were in line with those of RNA sequencing. Furthermore, it was discovered that QJL significantly upregulated the expression of p-AMPK and downregulated the expression of sterol regulatory element binding transcription factor 1 (SREBP-1c). In vitro, biochemical indicators and staining assays demonstrated that QJL-containing serum inhibited lipid accumulation and oxidative stress. The qRT-PCR and Western blot analysis revealed that QJL-containing serum markedly enhanced the expression of p-AMPK and carnitine palmitoyltransferase 1a (Cpt1a), while suppressing the expression of SREBP-1c, fatty acid synthase (Fasn), and acetyl-coenzyme A carboxylase 1 (ACC-1). However, CC inhibited the above pharmacological activities of QJL-containing serum. Additionally, (2S)-Liquiritigenin, Glycyrrhetinate, Isovitexin, Taxifolin, and Yohimbine were proved to be the key active components of QJL. CONCLUSION:QJL had the potential to be a therapeutic drug for ALD by activating the AMPK pathway, thereby regulating lipid metabolism and inhibiting oxidative stress.

Background: Polycystic ovary syndrome (PCOS) leads to persistent anovulation, hyperandrogenism, insulin resistance, and polycystic ovary, and is mainly characterized by menstrual disorders, and reproductive dysfunction. Angelica sinensis (Oliv.) Diels root has been used in many classical formulas of traditional Chinese medicine, and is commonly used to treat various gynecological diseases.Purpose: To investigate the protective effect of water extract of A. sinensis root (WEA) on PCOS rats, and the mechanism by RNA sequencing, and 16S rDNA sequencing. Methods: The PCOS rat model was established by letrozole combined with high-fat diet (gavage; 2 months), and treated with WEA (gavage; 2 g/kg, 4 g/kg or 8 g/kg; 1 month). To evaluate the therapeutic effect of WEA on PCOS rats, vaginal smear, hematoxylin-eosin staining, and biochemical indicators detection were performed. The rat ovarian tissue was analyzed by RNA sequencing, and the results were verified by qRT-PCR, and Western blot. 16S rDNA sequencing was used to analyze the gut microbiota of rats. Results: The results of the vaginal smear, and hematoxylin-eosin staining showed that WEA improved estrous cycle disorder, and ovarian tissue lesions. WEA (4 g/kg or 8 g/kg; 1 months) alleviated hormone disorders, insulin resistance, and dyslipidemia. RNA sequencing showed that WEA intervention significantly changed the expressions of 2756 genes, which were enriched in phosphatidylinositol3-kinase/phosphorylated protein kinase B (PI3K/AKT), peroxisome proliferator-activated receptor (PPAR), mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK), and insulin signaling pathways. 16S rDNA sequencing found that WEA increased the species diversity of gut microbiota, and regulated the abundance of some microbiota (genus level: Dubosiella, Bifidobacterium, Coriobacteriaceae (UCG-002), and Treponema; species level: Bifidobacterium animalis, Lactobacillus murinus, and Lactobacillus johnsonii). Conclusion: WEA regulated hormone, and glycolipid metabolism disorders, thereby relieving the PCOS induced by letrozole combined with high-fat diet. The mechanism was related to the regulation of PI3K/AKT, PPAR, MAPK, AMPK, and insulin signaling pathways in ovarian tissues, and the maintenance of gut microbiota homeostasis. Clarifying the efficacy and mechanism of WEA in alleviating PCOS based on RNA sequencing and 16S rDNA sequencing will guide the more reasonable clinical use of WEA.

Bergenin (BER), a natural component of polyphenols, has a variety of pharmacological activities, especially in improving drug metabolism, reducing cholestasis, anti-oxidative stress and inhibiting inflammatory responses. The aim of this study was to investigate the effects of BER on liver injury induced by isonicotinic acid hydrazide (INH) and rifampicin (RIF) in mice. The mice model of liver injury was established with INH (100 mg/kg)+RIF (100 mg/kg), and then different doses of BER were used to intervene. The pathological morphology and biochemical indicators of mice were detected. Meanwhile, RNA sequencing was performed to screen the differentially expressed genes and signaling pathways. Finally, critical differentially expressed genes were verified by qRT-PCR and Western blot. RNA sequencing results showed that 707 genes were significantly changed in the INH+RIF group compared with the Control group, and 496 genes were significantly changed after the BER intervention. These differentially expressed genes were mainly enriched in the drug metabolism, bile acid metabolism, Nrf2 pathway and TLR4 pathway. The validation results of qRT-PCR and Western blot were consistent with the RNA sequencing. Therefore, BER alleviated INH+RIF-induced liver injury in mice. The mechanism of BER improving INH+RIF-induced liver injury was related to regulating drug metabolism enzymes, bile acid metabolism, Nrf2 pathway and TLR4 pathway.

目的 通过生态适宜性区划研究,为我国南沙参资源可持续利用提供依据.方法 通过查阅资料和实地采集,收集南沙参共493个样点信息(包括轮叶沙参263个、沙参230个),综合55个生态因子,应用地理信息系统和最大熵模型分析南沙参的生态适宜性分布.结果 影响南沙参生态适宜性的主要环境因子是降水量、温度、坡度、海拔、土壤亚类,其适宜生境主要集中在江苏、安徽、浙江、贵州、湖南、湖北等地.结论 本研究结果可为合理选择优质南沙参种植地区提供依据.

From the perspective of market classification of Cnidii Fructus, this paper revealed the scientific connotation of evaluating the quality grade of Cnidii Fructus by its appearance traits. Thirty batches of Cnidii Fructus in different grades were selected as the research objects. The canonical correlation analysis and principal component analysis(PCA) were used to explore the measurement values of 15 appearance traits and intrinsic content indexes. The results of correlation analysis showed that except the aspect ratio, the 5 appearance trait indexes(length, width, 1 000-grain weight, broken grain weight proportion, and chroma) and 9 internal content indexes(the content of moisture, total ash, acid insoluble ash, osthole, imperatorin, 5-methoxy psoralen, isopimpinellin, xanthotoxin, and xanthotol) showed significant correlation to varying degrees. In addition, there was a significant positive correlation between the first typical variable U_1 composed of appearance traits and the first typical variable V_1 composed of internal content indexes(CR_1=0.963, P<0.01). The results of PCA showed that the classification results of appearance traits for 30 batches of Cnidii Fructus were consistent with the actual information of the samples. Under the same analysis conditions, 30 batches of Cnidii Fructus were reclassified by 9 groups of internal content indexes, and the analysis results were consistent. From the classification standard of the appearance traits of the system study, the statistical results of 6 appearance traits of Cnidii Fructus showed a correlation with grades. There was a good correlation between the appearance and the internal content of Cnidii Fructus, and the appearance quality effectively predicted the level of the internal content. There is a certain scientific basis for the quality classification of Cnidii Fructus by main appearance traits. Appearance classification can replace quality grading to realize the "quality evaluation through morphological identification" of Cnidii Fructus.

Effective treatment of liver fibrosis remains a challenging medical problem. Taraxasterol (TAR) has anti-inflammatory, anti-tumor and hepatoprotective effects. Studies have shown that TAR has good biological activity against liver injury induced by various factors. However, the anti-fibrotic effect of TAR and its mechanism are never clarified. The purpose of this study was to investigate the effects of TAR in liver fibrosis and to reveal its possible mechanism by RNA sequencing. Our results suggested that TAR attenuated CCl4-induced hepatocyte necrosis, inflammatory infiltration and ECM deposition. TAR inhibited the levels of ALT, AST, ALP, γ-GT, LN, HA, PC III and IV-C in serum and TNF-α, IL-6, IL-1β and MDA in liver. In addition, TAR increased the activities of SOD and GSH-Px in liver. RNA sequencing analysis of liver tissues revealed that CCl4 and TAR significantly altered 4,155 genes and 2,675 genes, respectively. TAR reversed changes in ECM-related genes. More specifically, TAR mediated the expression of genes related to the activation of the Hippo pathway, while inhibiting the expression of genes related to the activation of HIF-1α, TGF-β/Smad, and Wnt pathways. In the validation experiments, the qRT-PCR results showed that the expression levels of Yap1, Tead3, Hif1α, Vegfa, Tgfβ1, Want3a, and Ctnnb1 mRNA were consistent with the RNA sequencing results. The Western blot results showed that TAR inhibited the levels of TGF-β1 and p-Smad2. In addition, the results in vitro were consistent with those in vivo. Therefore, we concluded that TAR improved CCl4-induced liver fibrosis by regulating Hippo, HIF-1α, TGF-β/Smad and Wnt pathways.

目的 通过对甘肃省款冬花的生态适宜性评价,为其资源合理利用提供参考.方法 通过网络资料和实地采集,收集188份款冬花样本,综合55个生态因子,应用地理信息系统和最大熵模型分析款冬花在甘肃省的生态适宜性分布.结果 影响款冬花生态适宜性的主要生态因子分别是4月份降水量、海拔、12月份降水量、等温性、土壤有效水含量等级、土壤亚类、植被类型.其适宜性区划:定西中南部、陇南大部、临夏南部、平凉中部、天水大部、甘南东部边缘为适宜区,甘南东部、陇南东部边缘、定西北部、临夏北部、平凉东部和西部、庆阳南部、兰州西部为次适宜区,甘南中西部、庆阳北部、白银、武威、金昌、张掖、酒泉为不适宜区.结论 本研究结果与款冬花的产区一致,可为合理选择优质款冬花种植地区提供依据.

Scoparone (SCO) is known to have curative effect of alleviating liver injury. The purpose of this study was to observe the therapeutic effect and possible mechanism of SCO against high-fat diet (HFD) induced non-alcoholic liver disease (NAFLD) through in vivo experiments and RNA sequencing. Male Kunming mice were fed with HFD for 8 weeks to establish a mouse model of NAFLD, and SCO was used to treat NAFLD. Histopathology and biochemical indicators were used to evaluate the liver injury and the efficacy of SCO. RNA sequencing analysis was performed to elucidate the hepatoprotective mechanism of SCO. Finally, the differentially expressed genes of cholesterol synthesis and fatty acid (triglyceride) synthesis pathways were verified by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. The histopathological results showed that HFD could lead to significant steatosis in mice, while SCO could alleviate liver steatosis remarkably in NAFLD mice. The determination of biochemical indicators showed that SCO could inhibit the increased serum transaminase activity and liver lipid level induced by HFD. RNA sequencing analysis of liver tissues found that 2742 and 3663 genes were significantly changed by HFD and SCO, respectively. SCO reversed the most of genes involved in cholesterol synthesis and fatty acid (triglyceride) metabolism induced by HFD. the results of the validation experiment were mostly consistent with the RNA sequencing. SCO alleviated liver injury and steatosis in NAFLD mice, which may be closely related to the regulation of cholesterol and fatty acid (triglyceride) metabolism.