ABSTRACT Developing long‐wavelength‐activated nitric oxide (NO) donors has attracted increasing attention for achieving precise photo‐controlled NO release with deep tissue penetration and minimal phototoxicity, yet this remains a major challenge. Herein, we report a deep‐red light‐triggered NO‐releasing molecule (DBTBT‐NO) via a novel photoexcitation‐mediated electron transfer process to induce abundant peroxynitrite generation. The DBTBT‐NO can be activated by deep‐red light to release NO, which is facilitated by photoinduced intramolecular charge separation. Meanwhile, superoxide anions generated during photolysis rapidly react with NO to produce massive peroxynitrite in mitochondria, which induces tumor cell pyroptosis at a low light dose of 15 J/cm 2 and a low concentration of 0.4 µ m . The robust pyroptosis triggered by DBTBT‐NO effectively inhibits tumor growth and metastasis in mice with excellent biosafety. This is the first report on boosting peroxynitrite generation through long‐wavelength photolysis of benzisothiadiazole‐based N ‐nitrosanilines via a distinct mechanism. This work provides a new approach to fabricate unimolecular peroxynitrite photoinitiators with high pyroptosis induction efficiency, facilitating simplified and improved photoimmunotherapy.

Objective Naoxintong Capsule (NXT) is a core drug commonly used in clinical treatment of cardiovascular and cerebrovascular diseases in China, which is based on Buyang Huanwu Decoction. However, the protective effect and dose–effect relationship of NXT on myocardial ischemia–reperfusion injury (MIRI) are unclear, so this study aimed to evaluate the cardioprotective effect of NXT and its potential mechanism, and explore its optimal dosage. Methods Sprague-dawley male rats received trimetazidine (20 mg/kg), NXT (0.08, 0.16, 0.32, 0.64 and 1.28 g/kg), or an equal volume of saline for seven days prior to MIRI induction via intragastric administration. The MIRI model was established by occluding the left anterior descending coronary artery for a duration of thirty min, followed by a two-hour reperfusion period. The dose–effect relationship and related mechanism of NXT protecting MIRI were investigated by integration effect and dose–effect metabolomics which based on multi-indicator composite index method. Results NXT could protect MIRI by improving cardiac function, decreasing myocardial injury indexes, reducing myocardial hypertrophy and infarct area, and ameliorating structural abnormalities. Through integration effect, it was found that the dose–effect relationship showed a bidirectional correlation of first increasing and then decreasing, and the best protective effect was achieved at 0.32 g/kg, and the differential metabolites induced by MIRI could be reversed to the greatest extent in this dose. More importantly, NXT could protect MIRI by intervening in a variety of metabolites, such as stearoyl-CoA, linoleoyl-CoA, glucosamine 6-phosphate, myricetin, and 4-chlorocatechol. Conclusion In this study, the integration effect and dose–effect metabolomics based on the multi-indicator composite index method were used to illustrate the NXT’s effect on MIRI, and it was found that the potential molecular mechanism of NXT’s dose–response effects was associated with seven key metabolic pathways and the nuclear factor-κB (NF-κB)/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3)/cystein-asparate protease-1 (Caspase-1)/gasdermin D (GSDMD) signaling pathway. So this study revealed the effect from the dual perspectives of metabolites and targets. Furthermore, the optimal dose of 0.32 g/kg that this study discovered may provide a certain reference for the application of NXT in cardiovascular diseases. These findings revealed the dose–effect characteristics and related mechanisms of NXT in MIRI for the first time, and provided a new insight for the prevention and treatment of MIRI.

Aging is a social problem that people pay particular attention to. Oxidative stress is closely related to aging. Natural polysaccharides have numerous biological activities, and they show good application prospect in preventing oxidative stress and aging prospects. Herein, morphological analysis, molecular weight analysis, monosaccharide composition, methylation analysis, and NMR spectrum analysis were used to characterize the structure of ZJP-04M, an isolated homogeneous polysaccharide from the Ziziphus jujuba fruit. Results indicated that in ZJP-04M, the trunk chain was formed through the alternately connected homogalacturonan skeleton and rhamnogalacturonan-I domain and the branch chain was formed by arabinan and arabinogalactan. After feeding the polysaccharide to Caenorhabditis elegans, ZJP-04M was found to exert a certain paraquet-induced protective effect on the oxidative damage. Under normal conditions, ZJP-04M could significantly prolong the lifespan of C. elegans but exhibited no obvious effect on its reproductive capacity. We futher confirmed that ZJP-04M can play an antiaging role by regulating DAF-16. Based on these results, ZJP-04M can be used in antioxidation and antiaging applications.

ETHNOPHARMACOLOGICAL RELEVANCE:Taraxacum mongolicum Hand.-Mazz. is a well-known plant used both medicinally and as food, commonly used in traditional Chinese medicine prescriptions to alleviate benign prostatic hyperplasia (BPH). However, the material basis and molecular mechanisms of T. mongolicum alone in improving BPH remain unclear. In recent years, triterpenoids have been considered to be a key chemical constituents for T. mongolicum to exert its biological activity. AIM OF THE STUDY:To explore the therapeutic efficacy and underlying mechanism of total triterpenoids from T. mongolicum (TTM) and its active constituents against BPH. MATERIALS AND METHODS:The chemical components of TTM were determined using UPLC-QTOF-MS analysis. We established a testosterone propionate (TP)-induced rat model of BPH to assess the potential of TTM in vivo. Subsequently, network pharmacology was combined with experimental results from a TGFβ1-stimulated BPH-1 cell model to reveal the molecular mechanism of TTM. The main active ingredient (taraxasterol, TAR) of TTM was screened by evaluating its antiproliferative ability against BPH-1 and WPMY cells. Eventually, RNA-sequencing, RT-qPCR, immunofluorescence, and Western blotting were employed to elucidate the potential molecular targets and signalling pathways of TAR in BPH rats. RESULTS:TTM was mainly composed of ten pentacyclic triterpenoids and one phytosterol, including TAR, lupeol, β-amyrin, taraxerol, and their acetates. TTM ameliorated TP-induced BPH by decreasing androgen levels and repressing inflammatory responses and oxidative stress. Furthermore, TTM inhibited epithelial-mesenchymal transition (EMT) and extracellular matrix (ECM) deposition via impeding the TGFβ1/Smad signalling pathway in BPH-1 cells based on the network pharmacology. Among the main chemical components of TTM, TAR exerted the strongest antiproliferative activity in vitro, and inhibited the growth of BPH-1 and WPMY-1 cells in a concentration dependent manner. Importantly, TAR also reduced androgen levels and inflammatory responses to balance proliferation and apoptosis in BPH rats. Transcriptomic analysis showed that TAR attenuated collagen deposition in BPH by inhibiting ECM-receptor interaction pathway. In addition, TAR notably suppressed EMT and the TGFβ1/Smad signalling in BPH rats, as evidenced by reduced the protein levels of collagen I, a-SMA, Snail, TGFβ1, p-Smad2/Smad2, and p-Smad3/Smad3, alongside an increase in E-cadherin expression. CONCLUSIONS:TTM or TAR could effectively improve TP-induced BPH by suppressing androgen levels, inflammatory response, and EMT via the TGFβ1/Smad signalling pathway. These findings may present new therapeutic approachs for BPH in clinical settings. Notably, this study is the first to systematically elucidate the therapeutic mechanism of triterpenoids from T. mongolicum in treating BPH.

The Zingiberis Rhizoma − Jujubae Fructus herb pair (ZJHP) is a classic herb pair in traditional Chinese medicine. The herb pair shows the effect of dispelling cold, harmonizing the middle and improving gastrointestinal function, and is widely used for patients with stomach cold syndrome (SCS), stomachache and anemofrigid cold. The gingerols, shogaols, flavonoids and triterpenic acids are the important bioactive ingredients of ZJHP. However, few pharmacokinetic studies have been investigated in vivo for the above compounds. To comprehend the kinetics of active components and promote their curative application, a fast and sensitive ultra-high performance liquid chromatography coupled with mass spectrometry (UHPLC-MS/MS) method was established for simultaneous determination of 12 analytes in normal and SCS rats in this study. The results showed that the pharmacokinetic parameters (Cmax, Tmax, t1/2z, MRT0-t, AUC0-t and AUC0-∞) in SCS model were significantly different from those in normal rats. In addition, the pharmacokinetics of rats given ZJHP were also varied from single herb oral administration, especially in model condition. These results indicated that the in vivo processes of the above analytes changed under pathological conditions and the compatibility of the herb pair could significantly influence the absorption of active components, which might provide an insight and further supports for the clinical application of ZJHP.

Myocardial ischemia (MI) is a significant contributor to ischemic heart diseases like angina pectoris and myocardial infarction. Reactive oxygen species produced during MI can trigger lipid peroxidation, damaging cell structure and function. Salvia miltiorrhiza (SM) has been widely used clinically in the treatment of cardiovascular diseases. However, in the process of rooting, the aboveground parts of this plant are usually discarded by tons. To make better use of these plant resources, the phenolic acids extracted and purified from the aerial part of SM were studied and chemically transformed, and the potential protective effect and possible mechanism of salvianolic acids containing a higher content of salvianolic acid A on MI were obtained. The transformed products of SM stem-leaves total phenolic acids with 8.16 % salvianolic acid A showed a better protective effect on the isoproterenol (ISO)-induced acute MI injury rat model. It can improve ST segment changes and has good antioxidant, anti-inflammatory and anticoagulant effects. In addition, the dysbiosis of gut microbiota and the related metabolic levels of short chain fatty acids (SCFAs), phenylalanine and glycerophospholipids were improved. This was achieved by reducing the abundance of Bacteroides, Faecalibaculum, and L-phenylalanine levels. In addition, the abundance of probiotics in Butyricoccus, Roseburia, and norank_f_Eubacterium_coprostanoligenes_group, as well as the contents of propionic acid and isobutyric acid, LPCs and PCs were increased. In conclusion, total phenolic acids of SM stem-leaves showed protective effects against ISO-induced rats, especially the strongest effect after conversion, which is a new option for the prevention and treatment of MI.

Background: Osteoporosis has emerged as a significant global public health concern, predominantly affecting postmenopausal women. Its pathogenesis is intricate and the disease course is protracted, imposing substantial medical burden on both society and individuals. Objective: To investigate the effects of 2,3,5,4′-tetrahydroxystilbene-2-O-β-d-glucoside (TSG) and acetylated-TSG (Ac-TSG) on osteoblast viability, in vivo pharmacokinetics in rats and anti-osteoporotic effects in ovariectomized rat model. Methods: Ac-TSG was obtained by acetylation of TSG, and the purity and structure of Ac-TSG were determined by HPLC and 1H NMR. The effects of TSG and Ac-TSG on MC3T3-E1 cell viability, pharmacokinetics in rats, blood biochemical indexes of OVX model rats were detected. Hematoxylin–eosin (HE) staining was used to observe bone tissue morphology, tartrate-resistant acid phosphatase (TRAP) staining was used to observe osteoclast morphology, micro-CT was used to perform three-dimensional reconstruction of femur, and femur parameters were analyzed. Results: The structure of the compound is Ac-TSG and the purity is more than 98%. Both TSG and Ac-TSG could reduce the damage of oxidative stress to MC3T3-E1 cells and effectively improve the levels of Ca, P, ALP and BGP in serum of OVX rats. Compared with TSG, Ac-TSG has a longer action time in vivo and can improve the femoral structure, the number of trabecular bone and the number of osteoclasts in rats. Conclusion: Ac-TSG conferred protection to MC3T3-E1 cells against oxidative stress-induced damage and enhanced their bioavailability. Simultaneously, Ac-TSG ameliorated abnormal bone metabolism and mitigated bone microstructural changes in OVX rats, exhibiting a protective effect against osteoporosis.

Tumor-associated macrophages (TAMs) in non-small cell lung cancer (NSCLC) promote tumor cell metastasis by interacting with cancer cells. Ginsenoside Re is capable of modulating the host immune system and exerts anticancer effects through multiple pathways. Both AMPK and STING are involved in the regulation of MΦ polarization, thereby affecting tumor progression. However, whether there is a regulatory relationship between them and its effect on MΦ polarization and tumor progression is unclear. The aim of this study was to provide mechanistic evidence that ginsenoside Re modulates MΦ phenotype through inhibition of the AMPKα1/STING positive feedback loop and thus exerts an antimetastatic effect in NSCLC immunotherapy. Cell culture models and conditioned media (CM) systems were constructed, and the treated MΦ were analyzed by database analysis, RT-PCR, Western blotting, flow cytometry, and immunofluorescence to determine the regulatory relationship between AMPK and STING and the effects of ginsenoside Re on MΦ polarization and tumor cells migration. The effects of ginsenoside Re (10, 20 mg/kg/day) on TAMs phenotype as well as tumor progression in mice were assessed by HE staining, immunohistochemical staining, and Western blotting. In this study, AMPKα1/STING positive feedback loop in NSCLC TAMs induced M2 type polarization, which in turn promoted NSCLC cell migration. In addition, ginsenoside Re was discovered to inhibit M2-like MΦ polarization, thereby inhibiting NSCLC cell migration. Mechanistically, Re was able to inhibit the formation of the AMPKα1/STING positive feedback loop, thereby inhibiting its induction of M2-like MΦ and consequently inhibiting the epithelial-mesenchymal transition (EMT) process of NSCLC cells. Furthermore, in mouse models, Re was found to suppress LLC tumor growth and colonization by inhibiting M2-type polarization of TAMs. Our finding indicates that ginsenoside Re can effectively modulate MΦ polarization and thus play an important role in antimetastatic immunotherapy of NSCLC.

目的 建立龙生蛭胶囊的HPLC指纹图谱,结合化学模式识别方法进行质量评价.方法 采用HPLC法建立龙生蛭胶囊的指纹图谱,进行相似度评价,确定共有峰;对测定结果进行层次聚类分析和主成分分析,并结合正交偏最小二乘-判别分析对样品进行模式识别,以VIP值大于1为标准筛选影响龙生蛭胶囊质量的差异性成分.结果 19批龙生蛭胶囊样品的HPLC指纹图谱共标定了20个共有峰,相似度均大于0.95,指认了9个共有峰,分别为槲皮素、没食子酸、原儿茶酸、紫丁香苷、绿原酸、芍药苷、刺五加苷E、异嗪皮啶、毛蕊异黄酮葡萄糖苷.19批样品可分为2类;前4个主成分的累积方差贡献率为85.504％;以VIP值大于1为标准,筛选出8个主要峰,13、8、2(没食子酸)、3(原儿茶酸)、1(槲皮素)、14(芍药苷)、10、11号峰.结论 建立了龙生蛭胶囊更全面、系统的质量评价和分析方法,为龙生蛭胶囊的质量标准提高提供理论依据.

目的:研究甘松药材不同采收期挥发油及有效成分的质量差异.方法:参考2020版《中华人民共和国药典》水蒸气蒸馏法进行挥发油提取测定及有效成分提取,结合化学计量方法及中药指纹图谱技术建立不同采收甘松HPLC指纹图谱,采用相似度评价、距离分析及主成分分析进行综合评价.结果:甘松不同采收期指纹图谱共指认共有峰15个,挥发油聚类分析为2类,主成分综合评价中共标记3个主成分,HPLC指纹图谱相似度为0.500～0.993,主成分因子分析综合得分为0.6741.结论:建立甘松不同采收指纹图谱评价方法,确定最佳采收期.

目的 探讨前列舒通胶囊抗慢性非细菌性前列腺炎(CNP)的作用.方法 采用角叉菜胶诱导建立CNP 大鼠模型,随机分为模型组、普乐安片(阳性药)组和前列舒通胶囊低、中、高剂量组,另设假手术组,每组 10 只.给药 4 周后取材,HE染色观察大鼠前列腺组织形态变化,ELISA法检测血清和前列腺组织IL-6、IL-1β、TNF-α水平,Western blot法检测前列腺组织p-PI3K/PI3K、p-Akt/Akt、p-NF-κB/NF-κB蛋白表达.结果 与假手术组比较,模型组大鼠前列腺组织内有大量炎性细胞浸润和腺体增生,血清和前列腺组织IL-6、IL-1β、TNF-α水平升高(P＜0.01),前列腺组织p-PI3K/PI3K、p-Akt/Akt、p-NF-κB/NF-κB蛋白表达升高(P＜0.01);与模型组比较,普乐安片组及前列舒通胶囊各剂量组大鼠前列腺炎症及破坏程度减轻,血清和前列腺组织IL-6、IL-1β、TNF-α水平降低(P＜0.05,P＜0.01),前列腺组织p-PI3K/PI3K、p-Akt/Akt、p-NF-κB/NF-κB蛋白表达降低(P＜0.01).结论 前列舒通胶囊对慢性非细菌性前列腺炎大鼠具有较好的保护作用,其机制可能与抑制PI3K/Akt/NF-κB信号通路蛋白表达及炎性介质的分泌有关.

Objective To establish the fingerprints of Kelu Oral Liquid and multi-component quality control method.Methods The fingerprints were established and the similarity was evaluated by ultra-performance liquid chromatography（UPLC） and Traditional Chinese Medicine Fingerprint Similarity Evaluation System（2012 version）, and 6 components in the preparation were determined.Results The fingerprints were established with 10 batches of Kelu Oral Liquid by UPLC. At 250 nm, a total of 25 common peaks were marked, and 4 common peaks were identified. At 354 nm, 19 common peaks were marked, and 5 common peaks were identified. The similarity with the control spectrum was ＞0.99. The content ranges of isoliquiritin-apioside, isoliquiritin, baicalin, baicalein, wogonoside and ammonium glycyrrhizinate in 10 batches of Kelu Oral Liquid were 25.057-29.652, 26.404-32.072, 824.457-1 291.929, 319.541-557.251, 4.243-15.839, and 250.566-479.037 μg·mL -1 , respectively.Conclusion The UPLC fingerprints and multi-component determination method established are reliable and simple, which can lay a foundation for the study of pharmacodynamic substances of Kelu Oral Liquid in future.

Objective To study the bioequivalence of generic and original hydrochloride terazosin tablets in Chinese healthy subjects after single dose under fasting conditions. Methods A single-center, random, open, single-dose, two-preparations, double-period, crossover study was adopted. A total of 24 healthy adult male and female subjects were included in the random crossover administration. Single oral dose 2 mg of test and reference were taken, respectively. Plasma concentration of terazosin was determined by liquid chromatography tandem mass spectrometry. The main pharmacokinetic parameters were calculated by Phoenix WinNonlin 8.0 software. Results The main pharmacokinetic parameters of the test and reference preparations of hydrochloride terazosin tablets in the fasting group were as follows: C max were（53.79±9.77） and（57.62±14.18） ng·mL -1 , AUC 0-t were（519.90±179.49） and（551.99±143.15） ng·mL -1 ·h, AUC 0-∞ were（565.59±161.15） and（576.91±149.43） ng·mL -1 ·h, t max were 1.00（0.28, 2.02） and 0.75（0.50, 1.67） h, t 1/2 were（10.35±1.71） and（10.30±1.61） h. Under fasting conditions, the 90% confidence intervals of the main pharmacokinetic parameters of the test and reference preparations were in the range of 80.00%-125.00%. Conclusion Under fasting conditions,a single oral dose of generic and original hydrochloride terazosin tablets in Chinese healthy adult volunteers showed bioequivalence.

Yangzheng Mixture is a traditional Chinese medicine used in clinical practice as an adjuvant therapy for tumors. However, little is known about its active components in tumor treatment. The purpose of this study was to explore the potential anti-tumor components of Yangzheng Mixture to better promote its clinical application. Using LC-MS/MS, 43 components were detected in concentrated Yangzheng Mixture. Six components, comprising astragaloside, calycosin, formononetin, isoquercitrin, ononin, and calycosin-7-O-β-D-glucoside, were identified in rat plasma. The cancer cell absorption assay showed that the intracellular concentration of four components, calycosin, calycosin-7-O-β-D-glucoside, formononetin, and ononin, increased with extended incubation time and demonstrated potential anti-tumor effects. The MTT assay results confirmed that Yangzheng Mixture inhibited different tumor cells proliferation. Additionally, the colony formation assay, flow cytometry analysis and wound healing displayed that Yangzheng Mixture and a combination of four components could inhibit colony formation, arrest the cell cycle and impair cell migration of tumor cells, including HCT-116, MHCC-97L, MCF-7 and NCI-H1299. In summary, our study highlighted the plausible application of Yangzheng Mixture as a potential adjuvant treatment for tumors. Furthermore, it identified effective anti-tumor components and provided evidences for the further clinical application of Yangzheng Mixture.

目的:探究不同处理对黄精种子萌发和幼苗生长的影响,完善黄精种子有性繁育技术体系.方法:采用不同pH值的缓冲溶液、不同外源激素溶液和不同种子包衣剂,分别处理黄精种子,综合测定分析各处理对黄精种子萌发和幼苗生长影响.结果:酸性环境pH(4～6)中黄精种子的发芽势、发芽率、初生小球茎直径、须根数均高于碱性环境pH(8～10).不同外源激素对黄精种子萌发和幼苗生长均有显著的促进作用.其中,6-BA处理组黄精种子的发芽势最高为20.00％,ABA处理组黄精种子的发芽率最高为44.00％,幼苗初生小球茎直径和须根数由高至低依次均为:ABA>6-BA>GA3>2,4-D.不同包衣剂对黄精种子萌发和幼苗生长均有一定的抑制作用,其中SLS处理组对黄精种子萌发和幼苗生长的抑制作用最弱,且与CK组无显著差异(P<0.05).综合评分表明,对黄精种子萌发和幼苗生长促进作用较强的处理组分别为:pH4处理组、ABA处理组和SLS处理组.结论:酸性土壤环境有利于黄精种子萌发和幼苗生长,100 mg/L ABA溶液可以显著促进黄精种子萌发和幼苗生长,SLS包衣剂可以作为黄精种子防治病虫害的优选包衣剂.

目的 建立前列舒通胶囊的HPLC指纹图谱并结合化学模式识别,对前列舒通胶囊的整体质量进行评价.方法 采用HPLC法测得18批样品的色谱图,并进行相似度分析;以22个共有峰的峰面积为变量,进行聚类分析、主成分分析和正交偏最小二乘法-判别分析.结果 18批样品的指纹图谱中有22个共有峰,相似度均大于0.98,造成不同批次间的差异性成分为F3、F7、F16、F21等4个峰,其中,F16为盐酸小檗碱.结论 所用方法准确可行,可为"药材-中间体-成品"全过程的质量控制提供依据.

Objective: The inductively coupled plasma-mass spectrometry（ICP-MS） method was established for the simultaneous determination of 26 metal elements such as Al, As, B, Ba, Ca, and Cd in fresh Rehmanniae Radix samples.Further, the content of metal elements in the fresh Rehmanniae Radix samples processed with five different methods was compared. This study aims to provide a scientific basis for improving the quality evaluation system and screening the optimum processing method of fresh Rehmanniae Radix products. Methods: The fresh Rehmanniae Radix samples were prepared by spray drying, freeze drying, microwave drying, vacuum drying, and oven drying, respectively. After microwave digestion, ICP-MS was employed to simultaneously determine the content of 26 metal elements in the samples. The principal component analysis（PCA）, orthogonal partial least squares-discriminant analysis（OPLS-DA）, and hierarchical cluster analysis（HCA） were then performed. Results: The established method showed good linear relationship of the metal element content（r＞0.999）, with the lower limits of detection of 0.004-12.493 μg·L–1 and the precision meeting the requirement（RSD＜5%）. The fresh Rehmanniae Radix samples processed with different methods contained rich metal elements, with high content of K, Na, Ca, Mg, Al, Fe, and Zn. The metal element content varied in the fresh Rehmanniae Radix samples processed with different methods. The content of the macroelements K and Na and the microelements Fe, Zn, Se, Cu, Co,and Mo essential for the human body were higher in the sample processed with spray drying than in the samples processed with other methods. PCA classified 15 batches of fresh Rehmanniae Radix samples processed with different methods into 5groups. OPLS-DA extracted five metal elements with the contribution rate above 1 to the grouping of fresh Rehmanniae Radix samples processed with different methods, which were K, Al, Na, Mg, and Fe, respectively. HCA classified the 15samples into two groups, including a group of samples processed with freeze drying and microwave drying and a group of samples processed with oven drying, spray drying, and vacuum drying, and the samples processed with spray drying and vacuum drying were clustered as a sub-group. Conclusion: The method established in this study is simple, reliable, accurate,and sensitive and can be used for the determination of metal elements in the fresh Rehmanniae Radix samples processed with different methods. Considering the results of this study and the development trend and market demand of Chinese herbal medicine processing technology, we suggest that spray drying, which can maintain the color and rich metal elements and be industrialized, be selected as the optimum processing method of fresh Rehmanniae Radix. This study provides a research basis for improving the quality control system and the development and utilization of fresh Rehmanniae Radix.

Background: With the gradual increase in prevalence in recent years, non-alcoholic steatohepatitis (NASH) has become one of the significant health problems that urgently needs to be addressed worldwide. GanShuang Granules (GSG) is derived from the classical Chinese formula Xiaoyao San and mainly used in the clinical treatment of chronic liver diseases. Objective: In this study, we aim to gain a deeper insight into the inhibiting effects of GSG on non-alcoholic fatty liver disease (NAFLD) rats and preliminarily elucidate the underlying intervention mechanisms. Methods: First, High performance liquid chromatography (UHPLC-Q/Orbitrap-MS/MS) was used for the active compounds prediction in GSG. Then the data was mapped to mzCloud database. The targets corresponding to GSG compounds were collected from public databases, along with disease genes for NAFLD. The core targets and molecular mechanisms of GSG for NAFLD treatment were predicted by protein-protein interaction (PPI) network, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analyses. Molecular docking of the core target-component interactions was simulated using AutoDock Vina software. The effect of GSG on NASH rats was evaluated by pathological staining and analysis of various index results. Finally, the candidate targets were further validated by ELISA and western blot (WB) analyses. Results: Combining UHPLC-Q/Orbitrap-MS/MS data analysis and public database data, a total of 346 cross-targets were obtained, corresponding to 81 compounds. The subnetwork with an MCODE score of 53.623 is a potential core target group for this study. GO and KEGG enrichment analyses showed that the targets of GSG in NAFLD were mostly related to oxidative stress, the NF-κB signaling pathway, and the apoptosis signaling pathway. By integrating the results of network pharmacology analysis, the core objectives of this study mainly include AKT1, CASP9, TNF, and CASP8. The core ingredients are related to resveratrol and fisetin. The molecular docking results indicated key binding activity between AKT1-fisetin, AKT1-Resveratrol, and CASP8-fisetin. Moreover, GSG could improve the inflammatory status and restore the abnormal lipid accumulation of NAFLD/NASH liver, and these levels are further verified by pathological staining and detection of related indicators. Mechanistically, GSG could regulate protein expression levels in the liver for P65, p-P65, IKB, p-IKB, IKK, caspase-3, -8, -9, and cytochrome C, etc. It reflects the inhibitory effect of GSG on the NF-κB/IκB signaling pathway. Conclusion: Our results suggested that GSG demonstrated therapeutic effects on NAFLD/NASH rats, and these may be mainly reflected in the inhibitory effects on the NF-κB/IκB signaling pathway and its downstream inflammation and apoptosis signals.

目的 评价地氯雷他定片仿制药与原研药在中国健康受试者中单剂量空腹和餐后条件下给药的生物等效性.方法 用单中心、随机、开放、单次给药、两制剂、两周期、双交叉试验设计,空腹和餐后试验各入组24例受试者.空腹或餐后条件下单次口服地氯雷他定片受试制剂和参比制剂5 mg,用液相色谱串联质谱法测定血浆中地氯雷他定的浓度.用Phoenix WinNonlin 8.0软件计算主要药代动力学(PK)参数.结果 空腹组的地氯雷他定片受试制剂和参比制剂主要PK参数:地氯雷他定Cmax分别为(3.81±1.44)和(3.76±1.25)ng·mL-1,AUC0-t分别为(52.18±19.21)和(50.71±18.21)ng·mL-1·h,AUC0-∞分别为(54.52±19.71)和(53.19±19.07)ng·mL-1·h.餐后组的地氯雷他定片受试制剂和参比制剂主要PK参数:地氯雷他定Cmax分别为(3.52±1.20)和(3.55±1.10)ng·mL-1,AUC0-t分别为(64.33±52.76)和(61.59±42.24)ng·mL-1·h,AUC0-∞分别为(75.48±93.64)和(72.03±64.78)ng·mL-1·h.在空腹和餐后条件下,受试制剂与参比制剂主要PK参数的90％置信区间均在80.00％～125.00％内.结论 在空腹和餐后条件下,中国健康受试者单次口服地氯雷他定片仿制药与原研药具有生物等效性.