Introduction:Wuji Wan (WJW) is a classical Chinese formula traditionally prescribed for diarrhea/dysentery and abdominal pain. In ulcerative colitis (UC), inflammatory diarrhea reflects not only mucosal inflammation but also inflammation-linked disruption of epithelial electrolyte and water handling, highlighting membrane transport as a mechanistic bridge between symptom burden and immune activation. However, it remains unclear whether WJW confers therapeutic benefit in UC and whether any benefit is accompanied by coordinated regulation of membrane-transport-linked pathways. This study therefore asked whether WJW shows therapeutic effects in a UC model and whether these effects are accompanied by changes in epithelial Na+/Cl- transport and water-channel programs and by modulation of the T-cell-linked potassium channel Kv1.3. Methods:We investigated this question in a mouse model of DSS-induced colitis (3% dextran sulfate sodium). Our assessment included disease activity index (DAI) scores, histopathological analysis, ELISA, Western blotting, untargeted metabolomics, and whole-cell patch-clamp electrophysiology. Results:WJW significantly ameliorated DSS-induced colitis, as reflected by improved colonic pathology and partial normalization of DSS-associated serum metabolic perturbations. Untargeted metabolomics highlighted transport-related pathways. WJW increased/normalized the expression of key epithelial transport proteins involved in Na+/Cl- absorption and water handling, including sodium/hydrogen exchanger 3 (NHE3), epithelial sodium channel (ENaC), downregulated in adenoma (DRA), aquaporin-3 (AQP3), and aquaporin-8 (AQP8). In parallel, WJW reduced IL-6, IL-17A, and IFN-γ and dampened ERK/NF-κB pathway activation. WJW also reduced colonic Kv1.3 protein expression, and WJW-containing plasma directly inhibited Kv1.3 currents in Jurkat T cells. Conclusion:WJW ameliorated DSS-induced colitis and was accompanied by coordinated modulation of epithelial and immune membrane-transport-linked readouts.

This study aims to investigate the action mechanism by which Stellera chamaejasme extract(SCLE) overcomes multidrug resistance(MDR) in breast cancer through the mitogen-activated protein kinase kinase(MEK)/extracellular signal-regulated kinase(ERK) signaling pathway. Sensitive cell line MCF-7 of human breast cancer and its multidrug resistance counterpart MCF-7/adriamycin(ADR) were employed as models. The effect of SCLE on cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay. The transcriptional level of P-glycoprotein(P-gp) was analyzed by quantitative real-time PCR(qPCR). Western blot was employed to detect the expression levels of MEK and ERK, their phosphorylated forms(p-MEK and p-ERK), the drug resistance-related protein P-gp, DNA damage-related proteins including phosphorylated histone H2AX(γ-H2AX) and the tumor suppressor p53(p53), apoptosis-related proteins B-cell lymphoma-2(Bcl-2), Bcl-2-related X protein(Bax), and cleaved caspase-3. Immunofluorescence staining was used to observe the focus formation of DNA damage marker γ-H2AX, and flow cytometry was applied to detect apoptosis. Furthermore, DNA microarray was performed to identify the effect of SCLE on differentially expressed genes(DEGs) in MCF-7/ADR cells. Gene Ontology(GO) function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were conducted on these DEGs. The MTT results showed that the resistance factor(RF) of MCF-7/ADR cells in the SCLE group was 0.77, which was significantly lower than that observed for vincristine(VCR), paclitaxel(PTX), and ADR. Western blot and qPCR results indicated that SCLE inhibited the activation of the MEK/ERK pathway and significantly downregulated the expression of P-gp at both the protein and transcriptional levels(P<0.05). Concurrently, the SCLE treatment group increased the number of γ-H2AX foci and significantly upregulated the protein expression of p53, Bax, and cleaved caspase-3, while downregulating Bcl-2(P<0.05). Annexin-V/PI double staining revealed a dose-dependent increase in the apoptosis rate of the resistant cells(P<0.05). KEGG and GO analyses corroborated the cellular experimental findings, suggesting that SCLE may concurrently target multiple pathways, including adenosine triphosphate(ATP) binding, endoplasmic reticulum function, the mitogen-activated protein kinase(MAPK) pathway, the mammalian target of rapamycin(mTOR) pathway, and the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt) pathway, to coordinately induce DNA damage and apoptosis. In conclusion, SCLE may play a role in overcoming the MDR effect in breast cancer by inhibiting the activation of MEK/ERK in the core downstream cascade within the MAPK pathway, modulating P-gp levels to enhance intracellular drug accumulation, and subsequently triggering DNA damage and apoptosis.

IntroductionIrritable bowel syndrome (IBS) is a typical disorder of gut-brain interaction (DGBI). The microbiota-gut-brain (MGB) axis is pivotal in preventing and treating IBS. Wuji Pill is a traditional Chinese medicine commonly used to treat IBS. This study aimed to investigate the mechanism by which Wuji Pill improves IBS via the MGB axis.MethodsThe visceral sensitivity and colonic motor function were evaluated using the abdominal wall withdrawal reflex test and the colonic motility curve. Depression-like behavior were evaluated using sucrose preference test, open field test, novelty-suppressed feeding test, and forced swimming tests. The intestinal mucus secretion and the activation status of microglia was detected using AB-PAS staining and immunofluorescence staining, respectively. The species composition and abundance of gut microbiota were detected through 16S rRNA sequencing and RT-qPCR. Targeted metabonomics and RT-qPCR were used for metabolites and metabolic enzymes analysis.ResultsIn this study, Wuji Pill improved the symptoms of IBS rats and increased the relative abundance of Akkermansia muciniphila in feces. Additionally, antibiotics affected the repair of intestinal mucus secretion and significantly reduced the level of short-chain fatty acids. Subsequently, fecal microbiota transplantation and A. muciniphila transplantation can improve the symptoms of IBS rat by increasing intestinal mucus secretion, elevating the levels of acetic acid and butyric acid in feces. Additionally, the microglia in the cortex were suppressed, and the tryptophan-kynurenine pathway in the hippocampus was inhibited, leading to the conversion of tryptophan into 5-HT.DiscussionThis study highlights the Wuji Pill may alleviate IBS symptoms by modulating A. muciniphila and regulating the tryptophan metabolism pathway through MGB axis.

This study aimed to investigate the mechanism of dihydroartemisinin(DHA)in ameliorating multiple sclerosis(MS).Hematoxylin and eosin(HE)staining was used to assess inflammatory cell infiltration,while luxol fast blue(LFB)staining and electron microscopy were performed to evaluate myelin sheath structure.In cell experiments,this study measured programmed cell death ligand 1(PD-L1)expression on BV2 cells and forkhead box protein p3(Foxp3)expression in Jurkat T cells co-cultured with BV2 cells,determined the C-C motif chemokine ligand 5(CCL5)concentration in the supernatant of BV2 cells,and evaluated BV2 cell chemotaxis.Western blot(WB)was performed to detect protein levels of receptor tyrosine kinase(AXL),phosphorylated AXL(p-AXL),signal transducer and activator of transcription 1(STAT1),phosphorylated STAT1(p-STAT1),and suppressors of cytokine signaling 3(SOCS3).To confirm the role of AXL,key cellular assays were repeated following inhibition of AXL.Additionally,under physiological conditions,the effects of DHA on body weight,spleen weight,and peripheral blood immune cell profiles were examined.The results showed that DHA significantly reduced disease scores,attenuated body weight loss,suppressed inflammatory infiltration,and promoted myelin sheath repair in experimental autoimmune encephalomyelitis(EAE)mice.At the cellular level,DHA upregulated PD-L1 expression on BV2 cells and Foxp3 expression in co-cultured Jurkat cells,and inhibited CCL5 release and BV2 cell chemotaxis.It also upregulated AXL,p-AXL,p-STAT1,and SOCS3 protein expression in BV2 cells.When AXL was inhibited,these effects are nullified.In healthy mice,DHA did not have any effect on their various parameters.In conclusion,DHA maintains inflammatory homeostasis in the EAE model by activating the AXL signaling pathway in microglia.

This study employed the methods of serum chemistry,in vivo drug distribution analysis,and network pharmacology to investigate the potential therapeutic effects of Bufei Pills on respiratory disorders and decipher the underlying mechanisms.For the in vivo distribution study,rats were administrated with Bufei Pills,and the absorbed components were analyzed by HPLC-Q-TOF-MS/MS.The results showed that 56 components were present in the rat serum,including neocyclomorusin,asterinin A,neoisostegane,goimisin R,and frehmaglutoside H.These components were absorbed orally into the blood and distributed in various organs such as the heart,liver,spleen,lung,kidney,and pancreas as well as in the cerebrospinal fluid.The network pharmacological analysis identified 81 core targets such as ALB,IL6,AKT1,TNF,EGFR,ESR1,JUN,and HSP90AA1.These targets were involved in signaling pathways such as PI3K-AKT,MAPK,VEGFA,and AGE-RAGE.These findings suggested that Bufei Pills might be effective in treating respiratory disorders such as chronic obstructive pulmonary disease(COPD),pulmonary fibrosis,and pneumonia as well as COPD complications such as diabetes and depression by regulating inflammation,oxidative stress,metabolism,and immune function via the targets.In addition,Bufei Pills play a role in alleviating the shortness of breath,fatigue,and bodily pain during the recovery stage of COVID-19.Further research and development are needed to fully evaluate the potential of Bufei Pills.

This study aims to explore the effects of tetramethylpyrazine(TMP)on pharmacokinetics in plasma and brain dialysate and neuropathic pain in the rat model of partial sciatic nerve injury(SNI),and to investigate the correlation between the analgesic effect of TMP and its concentrations in the plasma and brain dialysate.Male SD rats were randomized into Sham,SNI,and SNI+TMP groups.Mechanical stimulation with von frey filaments and cold spray method were employed to evaluate the mechanical sensitivity and cold sensitivity of rats.Another two groups,Sham+TMP and SNI+TMP,were used to intubate the common jugular vein and implant microdialysis probes into the anterior cingulate gyrus(ACC),respectively.After intraperitoneal injection of TMP at a dose of 80 mg·kg-1,automatic blood collection and intracerebral microdialysis(perfusion rate of 1 μL·min-1)systems were used to collect the blood and brain dialysate for 24 h.HSS T3 C18 reversed-phase chromatographic column(2.1 mm×50 mm,2.5 μm)was used for liquid chromatographic separation.Gradient elution was carried out with the mobile phase of methanol-water(containing 0.005%formic acid)at a flow rate of 0.25 mL·min-1.Electrospray ion source was used for mass spectrometry,and the scanning mode was multi-reaction monitoring under the positive ion mode.The ion pairs for quantitative analysis were TMP m/z 137/122 and aspirin m/z 179/137,respectively.DAS 2.11 was used to calculate the pharmacokinetic parameters.The optimal time of TMP to exert the analgesia effect and inhibit cold pain sensitivity was 60 min after treatment.The TMP in the plasma and brain dialysate of SNI rats showed the Tmax of 15 min and 30 min,the Cmax of(2 866.43±135.39)and(1 462.14±197.38)μg·L-1,the AUC0-t of(241 463.30±28 070.31)and(213 115.62±32 570.07)μg·min·L-1,the MRT0-t of(353.13±47.73)and(172.16±12.72)min,and the CLZ of 0.73 and 0.36 L·min·kg-1,respectively.The analgesic effect of TMP had a significant correlation with the blood drug concentration in the ACC,which indicated that this method was suitable for the detection of TMP in rat plasma and brain dialysate.The method is accurate,reliable,and sensitive and can realize the important value of the application of correlation analysis theory of"automatic blood collection-microdialysis/PK-PD"in the research on neuropathic pain.

ETHNOPHARMACOLOGICAL RELEVANCE:Shenlian (SL) extract, a Chinese medicinal compound mainly pointing at inflammation response of atherosclerosis, is composed of Salvia miltiorrhizae Bunge and Andrographis paniculata (Burm.f.) Nees. Salvia miltiorrhizae Bunge has been reported to activate blood to remove stasis, while another herb, Andrographis paniculata (Burm.f.) Nees, has been revealed to clear endogenous heat toxins. The anti-atherosclerotic effects of these two herbs have been reported closely relating to inflammation. However, from the point of view of adventitial inflammation, the in-depth study of SL extract in anti-atherosclerotic effects by relieving adventitial inflammation is still unknown.AIM OF THE STUDY:To explore the effects of adventitial inflammation in atherosclerosis progression and if SL extract could reverse the process.MATERIALS AND METHODS:A novel atherosclerosis model based on adventitial inflammation was established. High-fat diet-fed ApoE-/- mice were implanted a cotton thread soaked with LPS on the right common carotid artery (RCCA). Meanwhile, three time points were set (week 2, 4, and 12) to accurately evaluate the effect of SL extract on the whole process of atherosclerosis with adventitial inflammation. The pathological changes of phenotype transformation of VAFs, vascular cell proliferation and collagen synthesis were observed dynamically by immunohistochemistry (IHC), BrdU method and sirius red staining. Then primary VAFs were stimulated by LPS to mirror the process of adventitial inflammation in vitro. The VAFS phenotype conversion and its function alterations including proliferation, migration, inflammatory secretion was assessed. Finally, we established a co-culture model of activated VAFs and vascular smooth muscle cells (VSMCs) to observe the impacts of activated VAFs on phenotype transformation and migration of VSMCs.RESULTS:SL extract improved atherosclerosis progression by reducing lipid content, adventitial inflammation and plaque formation. HE results showed sham-operated group (Sham) appeared light infiltrated inflammation only in adventitia at week 2, and the degree of inflammation infiltrated in model was more severe than that in Sham at week 2, 4, and 12. At week 12, the sham and model group showed evidently thickened media and intima. The phenotypic transformation, proliferation and migration of vascular adventitial fibroblasts (VAFs) as well as inflammatory secretion enhanced remarkably in vivo and vitro, but SL extract reversed these changes. Moreover, SL extract downregulated JAK2-STAT3-MMP2 signal pathway. The VSMCs transformed from contractile phenotype into synthetic phenotype and the migration of VSMCs increased after co-culture with activated VAFs. In contrast, SL extract could suppress theses effects.CONCLUSIONS:Taken together, atherosclerotic inflammation could be a "outside-in" signaling. Adventitial inflammation not only accelerated intimal plaque formation in atherosclerosis, but also worsened the degree of vascular lesion. And SL extract improved atherosclerosis by relieving adventitial inflammation, and the underlying mechanisms could be associated with curbing phenotypic transformation, proliferation and migration of VAFs and VSMCs.

This study investigated the chemical and biological activity of the secondary metabolites from an endophytic fungus Fusa-rium solani MBM-5 of Datura arborea. A total of six alkenoic acid compounds, including a new compound and five known ones, were isolated from the ethyl acetate extract of F. solani MBM-5 by using the chromatographic methods(open ODS column chromatography, silica gel column chromatography, Sephadex LH-20, and semi-preparative HPLC). The structures of the compounds were identified by using their physical and chemical data, spectroscopic methods(UV, IR, NMR, and HR-ESI-MS), and Mosher's reaction, which were fusaridioic acid E(1), fusaridioic acid C(2), fusaridioic acid A(3), L660282(4), hymeglusin(5), and hymeglnone(6). Compound 1 is new. MTT assay and Griss method were used to evaluate the growth inhibition of all the compounds against two tumor cells, as well as their influence and anti-inflammatory action on the release of NO from LPS-induced RAW264.7 cells. The results showed that compound 5 had strong growth inhibition activity against A549 and HepG2 cell lines, with IC_(50) values of 4.70 and 13.57 μmol·L~(-1), respectively. Compounds 1 and 6 significantly inhibited the release of NO from LPS-induced RAW264.7 cells, with IC_(50) values of 77.00 and 70.33 μmol·L~(-1), respectively.

Oral administration of Chinese Herbal Medicine (CHM) faces various challenges in reaching the target organs including absorption and conversion in the gastrointestinal tract, hepatic metabolism via the portal vein, and eventual systemic circulation. During this process, factors such as gut microbes, physical or chemical barriers, metabolic enzymes, and transporters play crucial roles. Particularly, interactions between different herbs in CHM have been observed both in vitro and in vivo. In vitro, interactions typically manifest as detectable physical or chemical changes, such as facilitating solubilization or producing precipitates when decoctions of multiple herbs are administered. In vivo, such interactions cause alterations in the ADME (absorption, distribution, metabolism, and excretion) profile on metabolic enzymes or transporters in the body, leading to competition, antagonism, inhibition, or activation. These interactions ultimately contribute to differences in the therapeutic and pharmacological effects of multi-herb formulas in CHM. Over the past two thousand years, China has cultivated profound expertise and solid theoretical frameworks over the scientific use of herbs. The combination of multiple herbs in one decoction has been frequently employed to synergistically enhance therapeutic efficacy or mitigate toxic and side effects in clinical settings. Additionally combining herbs with increased toxicity or decreased effect is also regarded as a remedy, a practice that should be approached with caution according to Traditional Chinese Medicine (TCM) physicians. Such historical records and practices serve as a foundation for predicting favorable multi-herb combinations and their potential risks. However, systematic data that are available to support the clinical practice and the exploration of novel herbal formulas remain limited. Therefore, this review aims to summarize the pharmacokinetic interactions and mechanisms of herb-herb or herb-drug combinations from existing works, and to offer guidance as well as evidence for optimizing CHM and developing new medicines with CHM characteristics.

Background:Jiaohong pills (JHP) consist of Pericarpium Zanthoxyli (PZ) and Radix Rehmanniae ,two herbs that have been extensively investigated over many years due to their potential protective effects against cognitive decline and memory im-pairment.However,the precise mechanisms underlying the beneficial effects remain elusive.Here,research studies were conducted to investigate and validate the thera-peutic effects of JHP on Alzheimer's disease. Methods:BV-2 cell inflammation was induced by lipopolysaccharide.AD mice were administered amyloid-β (Aβ).Behavioral experiments were used to evaluate learn-ing and memory ability.The levels of nitric oxide (NO),tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β),and interleukin-10 (IL-10) were detected using enzyme-linked immunosorbent assay (ELISA).The protein expressions of inducible nitric oxide synthase (iNOS) and the phosphorylation level of mitogen-activated protein kinase (MAPK) and nuclear factor kappa-B (NF-κB) were detected using Western blot.Nissl staining was used to detect neuronal degeneration. Results:The results demonstrated that an alcoholic extract of PZ significantly de-creased the levels of NO,IL-1β,TNF-α,and iNOS; increased the expression level of IL-10; and significantly decreased the phosphorylation levels of MAPK and NF-κB.These inhibitory effects were further confirmed in the AD mouse model.Meanwhile,JHP improved learning and memory function in AD mice,reduced neuronal damage,and enriched the Nissl bodies in the hippocampus.Moreover,IL-1β and TNF-α in the cortex were significantly downregulated after JHP administration,whereas IL-10 showed increased expression. Conclusions:It was found that JHP reduced neuroinflammatory response in AD mice by targeting the MAPK/NF-κB signaling pathway.

Objective To elucidate the pharmacodynamic substances and the molecular mechanism of the ancient classical formula Baoyuan Decoction(BYT),in order to explore more potential clinical application advantages of BYT.Methods The UPLC-QTOF-MS/MS technology was was used to perform gradient elution on a Waters ACQUITY UPLC-HSS T3 column(100 mm× 2.1 mm,1.7μm)with 0.1％formic acid water(A)-acetonitrile(B)as mobile phase system.High resolution time-of-flight mass spectrometry electrospray ionization source(ESI)and positive and negative ion scanning modes were used to detect and identify the chemical components of BYT,and the prototype chemical components and metabolites in the drug-containing plasma of BYT rats after multiple and multi-day ig administration were analyzed.Furthermore,key targeting effects and the therapeutic potential of the absorbed prototype components of BYT into blood were predicted using network pharmacological methods through TCMSP database and drug information database.Results According to accurate relative molecular mass data and multistage mass spectrometry fragment ions,combined with reference products,databases and literature reports,a total of 133 chemical components in BYT were identified,and it was determined that 35 components could be absorbed into the blood through oral absorption to achieve stable plasma concentration after rats were given BYT by ig.The"component-target"network map of the absorbed prototype components of BYT into blood,core target gene gene ontology(GO)functional enrichment analysis,Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis,and disease prediction analysis indicated that BYT might act on mitogen-activated protein kinase 1(MAPK1),tumor protein p53(TP53),signal transducer and activator of transcription and other core targets through eight key components,including ginsenoside Rb1,paeonol,formononetin,echinatin,daidzein,licochalcone A,6-gingerol and salicylic acid,and exert medicinal effects through the signaling pathways of advanced glycation end products-receptor for advanced glycation end-products(AGE-RAGE),epidermal growth factor receptor(EGFR),janus kinase-signal transducer and activator of transcription(JAK-STAT).It suggests that BYT has many potential therapeutic advantages on such as Alzheimer disease,reperfusion injury,postmenopausal osteoporosis,besides chronic heart failure.There were several derivatives of the same components in the 35 absorbed prototype components of BYT,,and one compound can dock with multiple targets,as well multiple analogues dock with the same target,which supporting the theoretical understanding of"multi-component,multi-target synergy of traditional Chinese medicine(formula)"and"multi-component,single-target superposition of traditional Chinese medicine".Conclusion Applying the UPLC-QTOF-MS/MS,network pharmacological and its database,which could provide a feasible scientifiic method for exploring the pharmacodynamic substances of traditional Chinese medicine(formula)and its molecular mechanism of action,as well clarifying the absorbed prototype components of BYT into blood and identifying some metabolites of some prototype components.And the target of action based on these prototype components was explained from the molecular level,and its potential therapeutic advantages were revealed,so as to provide reference for expanding the clinical application of this prescription and explore more clinical therapeutic potential.

Ethnopharmacological relevance: Shenlian (SL) extract is consisted of extracts from Salvia miltiorrhiza Bunge and Andrographis paniculata (Burm.f.) Nees, two herbs commonly used in Chinese clinical formula to treat atherosclerosis by removing blood stasis and clearing away heat. Pharmacologically, the anti-atherosclerotic effects of these two herbs are related to unresolved inflammation and the macrophage anergy or apoptosis in lesions led by the lipid flux blockage and ER stress. However, the deeper understanding of SL extract in protecting macrophage in plaques remains unknown.Aim of the study: This study aimed to investigate the underlying mechanism of SL extract in protecting ER-stressed macrophages from apoptosis in atherosclerosis.Methods: The ApoE-/- atherosclerotic mice model and ox-LDL loaded macrophages model were established to assess the effect of SL extract on ER stress in vivo and in vitro. Key markers related to ER stress in plaque were determined by immunohistochemical staining. Proteins involved in apoptosis and ER stress in macrophages loaded by ox-LDL were assessed by Western blot. ER morphology was observed by electron microscope. Lipid flux was temporally and quantitatively depicted by Oil red staining. The LAL and LXRa were blocked by lalistat and Gsk 2033 respectively to investigate whether SL extract protected the function of macrophages by the activation of LAL-LXRa axis.Results: Our study reported that, in ApoE-/- atherosclerotic mice, SL extract effectively relieved ER stress of carotid artery plaque. In lipid-overloaded macrophage models, SL extract significantly alleviated ER stress by promoting cholesterol degradation and efflux, which finally prevented apoptosis of foam cells induced by oxLDL. Blockage of ER stress by 4-Phenylbutyric acid (4-PBA), an inhibitor of Endoplasmic Reticulum (ER) stress, largely attenuated the protective effects of SL extract on macrophage. By utilizing the selective antagonists against both LAL and LXRa, this study further revealed that the beneficial effects of SL extract in macrophages was dependent on the proper functionalization of LAL-LXRa axis. Conclusions: By highlighting the therapeutic significance of macrophage protection in resolving atherosclerosis inflammation, our study pharmacologically provided convincing mechanistic evidence of SL extract in the activation LAL-LXRa axis and revealed its promising potential in the promotion of cholesterol turnover and prevention of ER stress induced apoptosis in lipid-loaded macrophages.

中枢神经系统(central nervous system,CNS)持续性的自身免疫性炎症和髓鞘脱失是多发性硬化症(multiple sclerosis,MS)的典型病理特征,尤其以神经系统持续衰退导致慢性脱髓鞘轴突病变引起的继发-进展型MS(secondary progressive multiple sclerosis,SPMS)最为难治.有多种免疫细胞和神经细胞参与MS的疾病进程,但在CNS中主要负责炎症反应调节和髓鞘再生的分别为小胶质细胞(microglia,MG)和少突胶质祖细胞(oligodendrocyte precursor cells,OPCs).在MS患者病灶部位发现具有MG M1/M2极化失衡和OPCs分化受阻导致的髓鞘再生障碍的现象,以往研究通常把MG的极化失衡和OPCs分化受阻当成两个独立的现象.但近期研究表明,两者之间存在交互式作用,MG极化失衡是导致OPCs分化受阻的核心机制之一,而OPCs也可反作用于MG,两者共同影响髓鞘的再生过程.在MS的治疗上,目前临床上治疗MS的药物只能对症治疗,无法根治MS.探索新的药物研发方向是目前MS临床治疗中亟待解决的关键科学问题.本研究就"从极化到分化"这一领域进行综述,聚焦于通过靶向MG极化失衡从而促进OPCs的分化和髓鞘再生的新策略,以期为MS及其他以CNS组织修复障碍为共同病理特征的神经退行性疾病的治疗提供新的药物研发方向.

The Wuji pill, also called Wuji Wan (WJW), is an effective traditional medicine for the clinical treatment of irritable bowel syndrome (IBS). It is principally composed of Rhizoma Coptidis, Fructus Evodiae Rutaecarpae, and Radix Paeoniae Alba. There have been no reports on the pharmacokinetics of WJW on IBS. Because it is more meaningful to study pharmacokinetics in relation to specific pathological conditions, our study investigated the pharmacokinetic differences of five representative components (berberine, palmatine, evodiamine, rutaecarpine, and paeoniflorin) in normal rats and chronic visceral hypersensitivity IBS (CVH-IBS) model rats after single dose and multiple doses of WJW using ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Transmission electron microscopy, immunohistochemistry, and immunofluorescence were used to explore mechanisms behind the pharmacokinetic differences in terms of tight junction proteins (Occludin and ZO-1), myosin light chain kinase (MLCK), and transporters including P-glycoprotein (P-gp), multidrug resistance associated protein 1 (MRP1), and multidrug resistance associated protein 2 (MRP2) in rat colons. After a single dose, for all components except rutaecarpine, significant differences were observed between normal and model groups. Compared with normal group, T1/2 and AUC0-t of berberine and palmatine in model group increased significantly (562.5 ± 237.2 vs. 1,384.9 ± 712.4 min, 733.8 ± 67.4 vs. 1,532.4 ± 612.7 min; 5,443.0 ± 1,405.8 vs. 9,930.8 ± 2,304.5 min·ng/ml, 2,365.5 ± 410.6 vs. 3,527.0 ± 717.8 min·ng/ml), while Cl/F decreased (840.7 ± 250.8 vs. 397.3 ± 142.7 L/h/kg, 427.7 ± 89.4 vs. 288.9 ± 114.4 L/h/kg). Cmax and AUC0-t of evodiamine in model group increased significantly (1.4 ± 0.6 vs. 2.4 ± 0.7 ng/ml; 573 ± 45.3 vs. 733.9 ± 160.2 min·ng/ml), while T1/2, Tmax, Cl/F, and Vd/F had no significant difference. Tmax and AUC0-t of paeoniflorin in model group increased significantly (21.0 ± 8.2 vs. 80.0 ± 45.8 min; 15,428.9 ± 5,063.6 vs. 33,140.6 ± 5,613.9 min·ng/ml), while Cl/F decreased (110.5 ± 48.1 vs. 43.3 ± 9.5 L/h/kg). However, after multiple doses, all five components showed significant differences between normal and model groups. Moreover, these differences were related to tight junction damage and the differential expression of transporters in the colon, suggesting that dose adjustment might be required during administration of WJW in the clinical treatment of IBS.

As a classic prescription, Wuji Pills is composed of Coptidis Rhizoma, Euodiae Fructus Preparata, and stir-fried Paeo-niae Radix Alba at the ratio of 6∶1∶6. The practical application of it is limited compared with other famous Chinese medicine prescriptions. Only one company produces Wuji Pills in China. In this study, ultra-performance liquid chromatography quadrupole time of flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to analyze and identify 26 identical compounds from Wuji Pills and drug-containing plasma of rats. Based on these components, 46 potential targets were screened out with network pharmacology methods, followed by the component-target network construction, Gene Ontology(GO) term enrichment, Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment, and disease prediction. It was concluded that Wuji Pills acted on core targets such as PTGS2, PTSG1, NCOA2, HSP9 OAD1, and RXRA through magnoflorine, hydroxyevodiamine, daucosterol, and berberine and exerted pharmacodynamic effects through various pathways such as calcium ion signaling pathway, phosphatidylinositol-3-kinase-protein kinase B(PI3 K-Akt) signaling pathway, and vascular endothelial growth factor(VEGF) signaling pathway. Thus, Wuji Pills has therapeutic potential for Alzheimer's disease, diabetes mellitus, myocardial ischemia, and other diseases in addition to the conventional disease(irritable bowel syndrome, IBS). The above research results can provide a reference for the comprehensive interpretation of the pharmacodynamic basis of Wuji Pills and the expansion of clinical application. At the same time, a lot of components in serum and the in vivo transformed and metabolized components of Wuji Pills have similar structure and relative molecular weight. In theory, these components may show additive effects and the competitive/antagonistic effects on the same target. According to the hypothesis of "additive effect of multiple components for a single target" in traditional Chinese medicine, multiple similar components may exert the additive effects on local targets. This study can partly prove the scientificity of this hypothesis and provide laboratory evidence.

目的 测定戊己丸中10个活性成分单次给药和多次口服给药后不同时间点的血药浓度,比较其在正常大鼠和慢性内脏高敏感肠易激综合征(CVH-IBS)大鼠体内药代动力学特征的差异.方法 采用乳鼠结肠球囊刺激法制备CVH-IBS大鼠模型,并对其进行内脏敏感性评价.单次或多次灌胃给予戊己丸提取物后于不同时间点从颈静脉采血,采用超高效液相色谱-串联质谱(UPLC-MS/MS)同时检测血浆中10个戊己丸活性成分的血药浓度,比较其药代动力学参数的差异.结果 CVH-IBS大鼠内脏敏感性增强.与单次给药相比,多次给药正常与模型大鼠达峰时间(tmax)均提前.单次给药后模型与正常大鼠的差异可正向验证前期实验结果.多次给药后与正常大鼠相比,模型大鼠体内戊己丸活性成分血药浓度峰值(Cmax)、曲线下面积(AUC0-t)、总清除率(Cl)发生显著改变.盐酸小檗碱、盐酸黄连碱、表小檗碱Cmax显著升高,盐酸巴马汀、盐酸药根碱、二氢小檗碱、吴茱萸碱、吴茱萸内酯Cmax显著下降;盐酸黄连碱、表小檗碱AUC0-t显著升高,盐酸巴马汀、二氢小檗碱、盐酸药根碱、吴茱萸碱、吴茱萸内酯AUC0-t显著下降;盐酸黄连碱、吴茱萸内酯Cl明显升高,表小檗碱Cl显著降低.盐酸巴马汀、芍药内酯苷t12明显降低;盐酸药根碱Vd显著升高.模型大鼠多次给药和单次给药相比较,黄连活性成分盐酸小檗碱、盐酸药根碱、表小檗碱、二氢小檗碱Cmax明显降低;盐酸药根碱Cl明显降低.白芍活性成分芍药苷t1/2明显下降,Cl明显上升.结论 戊己丸中活性成分在正常大鼠、CVH-IBS大鼠、戊己丸治疗后期的CVH-IBS大鼠体内的药代动力学行为存在明显差异,这可能与IBS治疗早期肠道屏障被破坏和治疗后期肠道屏障修复、药物肝肠循环蓄积作用以及肝酶的活性等代谢功能改变相关.

Context Shenlian extract (SL) is a combination of Salvia miltiorrhiza Bge. (Labiatae) and Andrographis paniculata (Burm. F.) Wall. Ex Nees (Acanthaceae) extracts, which promote blood circulation and clear endogenous heat toxins. Myocardial ischaemia-reperfusion injury (MI/RI) is aggravated myocardial tissue damage induced by reperfusion therapy after myocardial infarction. Objectives This study explores the effect of SL on MI/RI and the underlying mechanism. Materials and methods Primary peritoneal macrophages (pMACs) were treated with LPS and SL (5, 10 or 20 mu g/mL) for 24 h. The myocardial ischaemia-reperfusion (MI/R) model was established after administration of different doses of SL (90, 180 or 360 mg/kg). Myocardial tissue injury was assessed by methylthiazolyl tetrazolium (TTC) staining and levels of creatine kinase (CK), lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in mice. The double immunofluorescence staining of iNOS/F4/80 and CD86/F4/80 was used to detect macrophage M1 polarization. The levels of miR-155, inflammatory factors and chemokines were detected by qRT-PCR or ELISA. CD86, iNOS, SOCS3, JAK2, p-JAK2, STAT3 and p-STAT3 proteins expressions in macrophages were analyzed by western blotting. Conditioned medium transfer systems were designed to unite M1 macrophages with H/R cardiomyocytes, and cell apoptosis was detected by TUNEL staining, western blotting or immunohistochemistry. Results SL reduced apoptosis, diminished CK and LDH levels, raised SOD concentration and decreased infarct size in the MI/R model. Meanwhile, SL decreased miR-155 level, inhibited M1 macrophage polarization and inflammation. Furthermore, SL promoted SOCS3 expression and blocked JAK2/STAT3 pathway in vitro. Conclusions SL may be a promising TCM candidate for MI/RI. The underlying mechanisms could be associated with inhibition of M1 macrophage polarization via down-regulating miR-155.

To the Editor: In breast cancer, it is the metastasis that makes a clear distinction between life and death. However, effective treatment targeted against metastasis remains an unmet clinical need. Accumulating evidence suggests that an essential step in metastasis is tumor cell acquisition of motility. Furthermore, epithelial– mesenchymal transition (EMT) initiates the early steps of tumor cell spread by endowing them with greater motility and invasiveness. Focal adhesion kinase (FAK) controls cell mesenchymal characteristics responsible for invasiveness and adhesion, placing itself as a critical molecule in regulating EMT in various cancer cells.

Background: Non-small cell lung cancer (NSCLC) is a major subtype of lung cancer with a higher mortality rate. Both apoptosis and autophagy are crucial processes in the pathophysiology of NSCLC. Muyin extract (MSE) is a combination of Momordica cochinchinensis (Lour.) Spreng seeds and Epimedium brevicornu Maxim extract, with an optimal ratio of 1:1. Our previous research has firstly shown that MSE exerts a good anti-tumor activity, especially for NSCLC.Purpose: This study aims to evaluate the inhibitory effect of MSE on NSCLC and explore the underlying mechanism.Methods: In vitro , cell proliferation was examined by MTT and colony formation. Apoptosis was detected by annexin V-FITC/PI assay while autophagy was assessed by Acridine orange (AO) and Monodansylcadaverine (MDC) staining. In vivo , Lewis lung cancer cell transplanted mice model was established to measure the effect of MSE on tumor growth. Hematoxylin eosin (H & E) staining was used to observe the pathological changes of the tumor after MSE treatment. The apoptosis in tumor tissue was detected by TUNEL assay. Meanwhile, the cellular proliferation marker Ki67 and autophagy marker LC3II were observed by immunohistochemistry staining. The IL-4 and IFN-gamma concentrations in blood were tested by Elisa. The apoptosis related factors (Bcl-2, Bax Caspase-3, cleaved Caspase-3, Caspase-9 and p53), autophagy marker proteins (Atg-5, Becline-1, LC3II/Iand p62) as well as Akt/mTOR pathway were detected by western blotting.Results: Present study showed that MSE greatly inhibited the proliferation of NSCLC in vitro and in vivo , together with apoptotic rate increasing. P53 and cleaved Caspase-3 levels were up-regulated while Bcl-2/Bax ratio, Caspase-3 and Caspase-9 levels were significantly down-regulated treated with MSE. Meanwhile, MSE activated autophagy, Atg-5, Becline-1 as well as the ratio of LC3II/I were notably up-regulated while p62 was down regulated after MSE treatment. Importantly, MSE significantly blocked Akt/mTOR pathway, which is a common upstream signal triggered by autophagy and apoptosis. Furthermore, when co-treated with specific autophagy inhibitor, the inhibitory rate and anti-apoptotic Bcl-2 level were significantly reversed. Impressively, MSE remarkably increased IFN-gamma/ IL-4 ratio while VP16 did not in animal model, and the inhibition rate in tumor weight after MSE treatment was higher than xiaojin pill.Conclusion: Taken together, it is proved that MSE may be a promising oral TCM candidate for NSCLC therapy with immunity improvement. The underlying mechanisms could be associated with the induction of apoptosis and autophagy through blocking Akt/mTOR pathway, meanwhile, it may promote crosstalk between autophagy and apoptosis.