Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive cognitive impairment, and there remains a lack of effective treatments capable of reversing or significantly slowing disease progression. Accumulating evidence indicates that the immune function of microglia, the resident immune cells of the central nervous system, is a critical factor in regulating AD pathogenesis. Emerging research in immunometabolism further reveals that glucose metabolic reprogramming serves as a central driver of microglial phenotypic and functional differentiation. This review systematically outlines the fundamental characteristics of microglial glucose metabolism and focuses on how the dynamic metabolic reprogramming it undergoes during AD progression regulates microglial immune behavior and inflammatory responses. Building on this, we further summarize key regulatory targets within the "metabolism-immune axis" and corresponding pharmacological intervention strategies. Finally, this article discusses current challenges and future research directions in the field of microglial immunometabolism. This review aims to provide a theoretical foundation for AD intervention strategies targeting the "metabolism-immune axis" and to offer insights for the development of novel disease-modifying therapeutics.

Background Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovial inflammation, bone and cartilage damage, musculoskeletal pain, swelling, and stiffness. Inflammation is one of the key factors that induce RA. Jingfang Granule (JFG) is a traditional Chinese medicine (TCM) with significant anti-inflammatory effects. Clinical studies have confirmed that JFG can be used to treat RA, but the mechanism is still vague. Purpose This study was designed to evaluate the protective function and the mechanism of JFG on rats with RA. Study design and methods Complete Freud’s Adjuvant (CFA) was used to establish a rat RA model, and JFG or Diclofenac Sodium (Dic) was orally administered. Foot swelling and hematoxylin eosin (H&E) staining were used to test the therapeutic effect of JFG on RA treatment, while ELISA kits were used to detect serum cytokines. Malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), catalase (CAT), and reactive oxygen species (ROS) were used to evaluate oxidative stress levels. The integration of label free proteomics, fecal short chain fatty acid (SCFA) targeted metabolomics, peripheral blood SCFA, medium and long chain fatty acid targeted metabolomics, and 16S rDNA sequencing of gut microbiota were used to screen the mechanism. Western blot technology was used to validate the results of multiple omics studies. Serum D-Lactic acid, lipopolysaccharide specific IgA antibody (LPS IgA), diamine oxidase (DAO), and colon Claudin 5 and ZO-1 were used to evaluate the intestinal barrier. Results The results confirmed that JFG effectively protected rats from RA injury, which was confirmed by improved foot swelling and synovial pathology. At the same time, JFG reduced the levels of TNF-α, IL-1β, and IL-6 in serum by inhibiting the NLRP3 inflammasome signaling pathway and TLR4/NF-κB signaling pathway in synovial tissue. Multiple omics studies indicated that JFG increased the abundance of gut microbiota and regulated the number of gut bacteria, thereby increased the levels of Acetic acid, Propionic acid, and Butyric acid in the gut and serum of RA rats, which activated AMPK to regulate fatty acid metabolism and fatty acid biosynthesis, thereby inhibited lipid oxidative stress induced ferroptosis to improve tissue damage caused by RA. Meanwhile, JFG improved the intestinal barrier by upregulating the expresses of Claudin 5 and ZO-1, which was confirmed by low concentrations of D-Lactic acid, LPS-SIgA and DAO in serum. Conclusions This study confirmed that JFG improved the disturbance of fatty acid metabolism by modulating gut microbiota and the production of fecal SCFAs to activate AMPK, and then inhibited ferroptosis caused by lipid oxidative stress in synovium tissue and prevented AR injury. This study proposes for the first time to investigate the mechanism of JFG treatment for RA from the perspective of the "Gut-joint" axis, and provides a promising approach for the treatment of RA.

ETHNOPHARMACOLOGICAL RELEVANCE:Cerebral ischemic stroke (CIS) is one of the most important factors leading to death and disability, which seriously threaten the survival and health of patients. The intentional flora and its derived metabolites are demonstrated to play vital roles in the physiology and onset of CIS. Shouhui Tongbian Capsules (SHTB), a Traditional Chinese Medicine, could regulate gut microbiota and metabolites. Study has found that SHTB has protective effect on CIS, but the mechanism is still unclear. AIM OF STUDY:This study was designed to evaluate the preventive effects and the mechanism of SHTB on CIS injury. MATERIALS AND METHODS:The rats were pretreated with SHTB for 5 days, then the middle cerebral artery occlusion/reperfusion (MCAO/R) was established. Neurological deficit score, TTC staining, brain water content, H&E and Nissl staining were preformed to evaluate the preventive effects of SHTB on CIS. The Occludin and ZO-1 were analyzed to evaluate the blood-brain barrier (BBB). 16S rDNA sequencing and LC-ESI-MS/MS-based metabolomics profiling were performed to analyze the gut microbiota composition and short chain fatty acids (SCFAs) profile in gut. Serum lipopolysaccharide specific IgA antibody (LPS-SIgA) and diamine oxidase (DAO), as well as colon Claudin 5 and ZO-1 were analyzed to evaluate the intestinal barrier. Proteomics was used to evaluated the proteins profile in brain. Lipidomics were used to evaluate the brain SCFAs as well as medium and long chain fatty acids (MCFAs and LCFAs). Malondialdehyde (MDA), Total Superoxide dismutase (T-SOD), Glutathione (GSH), Glutathione peroxidase (GSH-Px), Catalase (CAT) and reactive oxygen species (ROS) were assayed to evaluate the oxidative stress in brain. Western blot was performed to evaluate the expression of PPARγ, Nrf2, SLC3A2, SCL7A11, GPX4, ACSL4 and LOX. RESULTS:SHTB prevented rats from MCAO/R injury, which was confirmed by lower cerebral infarct rate, brain water content, neurological deficit score and nissl body loss, and improved brain pathology. Meanwhile, SHTB upregulated the expression of ZO-1 and Occludin to maintain the integrity of BBB. 16S rDNA sequencing and LC-ESI-MS/MS-based targeted metabolomics found that SHTB increased the abundance of gut microbiota, regulated the numbers of intestinal bacteria to increase the production of Acetic acid, Propionic acid, and Butyric acid, as well as decrease the production of Valeric acid and Hexanoic acid in the gut. Meanwhile, SHTB improved the intestinal barrier by upregulating the protein levels of Claudin 5 and ZO-1, which was confirmed by low concentrations of LPS-SIgA and DAO in serum. Multi omics and spearman correlation analysis indicated that SHTB regulated the abundance of Escherichia-Shigella and Lactobacillus to increase Acetic acid, Propionic acid, and Butyric acid to induce the expression of PPARγ, thereby regulating fatty acid metabolism and degradation, improving lipid metabolism disorders, downregulating lipid oxidative stress, inhibiting ferroptosis, and alleviating brain injury. CONCLUSION:This study confirmed that SHTB improved the disturbance of fatty acid metabolism in brain tissue by regulating gut microbiota and the production of fecal SCFAs to inhibit ferroptosis caused by lipid oxidative stress and prevent CIS injury, which provided a potential candidate drug for the prevention of CIS.

In recent years, the gut microbiota has been increasingly recognized for its influence on various central nervous system diseases mediated by microglia, yet the underlying mechanisms remain unclear. As key metabolites of the gut microbiota, short-chain fatty acids (SCFAs) have emerged as a focal point in understanding microglia-related interactions. In this review, we further refine the connection between the gut microbiota and microglia by introducing the concept of the “SCFAs-microglia” pathway. We summarize current knowledge on this pathway, recent discoveries regarding its role in neurological diseases, and potential pharmacological strategies targeting it. Finally, we outlined the current challenges and limitations in this field of research. We hope this review provides new insights into the role of the gut microbiota in neuroimmune regulation.

Ethnopharmacological relevance Chronic fatigue syndrome (CFS), as a complex, multisystemic, and multisystemic disorder affecting multiple organs and systems, often accompanies by symptoms such as post-exercise discomfort, sleep disorders, cognitive difficulties, and orthostatic intolerance. Jingfang Granule (JFG) is a traditional Chinese medicine that have significant protective effects on CFS, but the mechanism is still vague. Aim of study This study was designed to evaluate the protective mechanism of JFG on mice with CFS. Materials and methods The combined stimuli method was used to establish the mice CFS model, and JFG was orally administered. The body weight, exhaustion swimming training and tail suspension test were assayed every 7 days to evaluate the improvement of JFG on CFS. Lactic acid, adenosine triphosphate (ATP), malondialdehyde (MDA), superoxide dismutase (SOD), reactive oxygen species (ROS), IL-1 beta, TNF-alpha, IL-6 in serum and liver glycogen, muscle glycogen in muscle were analyzed. Transmission electron microscopy was used to detect mitochondrial morphology. The regulatory networks were investigated by proteomics and central carbon metabolomics, which were verified by Western blot. Results JFG reversed the loss of weight and reduce of exhaust swimming time (P < 0.05) induced by CFS in mice, and increased the tail suspension time (P < 0.05), indicating that JFG has an improving effect on CFS. Meanwhile, JFG increased the spleen index (P < 0.05), decreased the thymus index (P < 0.05) and cardiac index (P < 0.05), inhibited the secretion of Lactic acid (P < 0.05), and increased the content of liver glycogen (P < 0.05), muscle glycogen (P < 0.05), and ATP (P < 0.05), and improved mitochondrial morphology in mice with CFS. JFG also inhibited the release of TNF-alpha (P < 0.05), IL-1 beta (P < 0.05) and IL-6 (P < 0.05) in serum by inhibiting TLR4/NF-kappa B signaling pathway and NLRP3 inflammasome signaling pathway, and inhibited oxidative stress by activating Nrf2/HO-1/NQO1 axis. Integrated central carbon metabolomics, proteomics and Western blot showed that JFG intervened in CFS by increasing the expression of Idh1 (P < 0.05) and Idh2 (P < 0.01) to promote tricarboxylic acid (TCA) cycle. Conclusions This study confirmed that JFG promoted the TCA cycle by increasing the expression of Idh1 and Idh2, and then inhibited inflammation and oxidative stress to prevent CFS injury, which provided a potential drug candidate for CFS treatment.

ETHNOPHARMACOLOGICAL RELEVANCE:Cervical spondylosis (CS) is a common condition primarily caused by intervertebral disc degeneration (IVDD), adversely affecting quality of life. Traditional Chinese medicine believes that Qi deficiency and blood stasis are the main pathogenesis of CS. Guiqi Huoxue capsule (GQHX) has the effect of beneficial Qi tonifying kidneys and promoting blood circulation, which is mainly used in the clinical treatment of CS (Qi deficiency and blood stasis syndrome). However, the underlying mechanism of action has not been reported and clarified. AIM OF THE STUDY:The aim of this study was to investigate the efficacy and underlying mechanisms of GQHX in CS rats. MATERIALS AND METHODS:The CS rat model (Qi deficiency and blood stasis syndrome) was established by using IVDD and Ovariectomy (OVX) surgeries, along with qi deficiency and blood stasis modeling. The effects of GQHX on CS rats were evaluated by behavioral tests, blood indexes, H&E staining, and other means. Fatty acid profiles and gut microbiota were analyzed using lipidomics and 16S rRNA sequencing. The mechanism of action of GQHX was investigated by network pharmacology and western blotting. RESULTS:GQHX reduced the symptoms of CS rats as confirmed by behavioral indicators, serum markers, and other measures of efficacy. Meanwhile, 16S rRNA sequencing and lipidomics results showed that GQHX regulated the abundance of Blautia and Muribaculaceae, influencing the production of various fatty acids (e.g. isobutyric, isovaleric, and linoleic acids). More importantly, network pharmacology and Western blot results suggested that GQHX could alleviate the clinical symptoms of CS by regulating the abnormal expression of AGE-RAGE, MAPK, and HIF-1 signaling pathways. CONCLUSION:This study elucidated the role of GQHX in alleviating CS and highlighted the mechanisms involved, particularly the regulation of gut microbiota and lipid metabolism, as well as the AGE-RAGE, MAPK, and HIF-1 signaling pathways.

This study aims to investigate the protective effect and potential mechanism of Jingfang Granules(JF) on the mouse model of chronic fatigue syndrome(CFS). Mice were randomized into normal, model, and low-, medium-, and high-dose(0.9, 1.8, and 3.6 g·kg~(-1)·d~(-1), respectively) JF groups according to the body weight. In addition to the normal group, other groups of mice received exhaustive swimming training and tail suspension training every day for the modeling of CFS. The mice in each administration group were administrated with JF at the corresponding dose by gavage, and those in the other groups were administrated with an equal amount of purified water. The exhaustive swimming and tail suspension tests were conducted in each group. The UV-glutamate dehydrogenase method was used to determine the serum level of urea nitrogen(UREA), and the lactate dehydrogenase(LDH) assay kit was used to determine the LDH level. Enzyme-linked immunosorbent assay was employed to measure the levels of interleukin-6(IL-6) and tumor necrosis factor-α(TNF-α) in the serum, muscle tissue, and brain tissue of mice in each group. Western blot was employed to determine the expression levels of Toll-like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), nuclear factor-kappa B(NF-κB) and their phosphorylated proteins in the muscle tissue of mice. The 16S rDNA sequencing and ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) were adopted to detect the changes of intestinal flora and intestinal metabolites in mice. Compared with the model group, JF significantly prolonged the swimming exhaustion time and shortened the tail suspension time of the model mice, lowered the levels of LDH and UREA in the serum as well as the levels of IL-6 and TNF-α in the serum, muscle tissue, and brain tissue of CFS mice. In addition, JF down-regulated the expression of TLR4, MyD88, and p-NF-κB/NF-κB in the muscle tissue of CFS mice compared with the model group. The results of 16S rDNA sequencing demonstrated that JF ameliorated the intestinal flora disorder of CFS mice. The results of UPLC-MS/MS revealed that JF significantly affected the histidine metabolism pathway in the intestinal tract of CFS mice. Spearman analysis displayed that histamine, a metabolite involved in histidine metabolism, was negatively correlated with the abundance of Clostridia＿UCG-014, Dubosiella, and RF39 and positively correlated with the abundance of Coriobacteriaceae＿UCG-002. The metabolite imidazole-4-acetaldehyde was negatively correlated with the abundance of Clostridia＿UCG-014, Dubosiella, and RF39 and positively correlated with the abundance of Coriobacteriaceae＿UCG-002. In conclusion, JF can increase the swimming exhaustion time, reduce the immobility time of tail suspension, lower serum LDH and UREA levels, and alleviate inflammation response. It may exert the therapeutic effect by improving intestinal flora homeostasis and inhibiting histidine metabolism by down-regulating the expression of proteins in the TLR4/MyD88/NF-κB signaling pathway, thereby relieving the symptoms of CFS in mice.

Ulcerative colitis (UC) is a chronic, relapsing nonspecific intestinal inflammatory disease. It is difficult for a single drug to treat UC effectively and maintain long-term efficacy. There is an urgent need to find new drugs and treatment strategies. MAGL11 is a new kind of single acylglycerol lipase (MAGL) inhibitor. Icaritin (Y003) is the major metabolite of icariin in vivo. Several studies have confirmed the role of MAGL inhibitors and icariin in anti-inflammatory and regulation of intestinal stability. Therefore, this study adopted a new strategy of combining MAGL inhibitor with Icaritin to further explore the role and mechanism of drugs in the treatment of UC. Enzyme-linked immunosorbent assay (ELISA), hematoxylin-eosin staining (HE), immunohistochemical (IHC) and Western blot were used to detect the synergistic protective effects of MAGL11 and Y003 on intestinal pathological injury, intestinal mucosal permeability and inflammation in UC mice. 16S rDNA sequencing was used to detect the synergistic effect of MAGL11 and Y003 on gut microbiota. The effects of MAGL11 and Y003 combined therapy on serum and fecal metabolism of UC mice were analyzed by untargeted metabolomics. Proteomics method was applied to investigate the molecular mechanisms underlying MAGL11 and Y003 synergy in the treatment of UC. The results showed that MAGL11 and Y003 could synergistically improve the clinical symptoms, reduce intestinal inflammation and pathological damage, and improve intestinal mucosal permeability in UC mice. The mechanism study found that MAGL11 and Y003 could synergistically inhibit Toll-like receptors 4 (TLR4) / Myeloid differentiation primary response gene (Myd88)/Nuclear factor kappa-B (NF-κB) pathway and further regulate gut microbiota imbalance and metabolic disorders to treat UC.

ETHNOPHARMACOLOGICAL RELEVANCE:Mai Men Dong decoction (MMDD), a traditional Chinese medicine formula, is relevant to ethnopharmacology due to its constituents and therapeutic properties. The formula contains herbs like Ophiopogon japonicus (Thunb.) Ker Gawl., Pinellia ternata (Thunb.) Makino, Panax ginseng C.A.Mey, Glycyrrhiza uralensis Fisch, and Ziziphus jujuba Mill, Oryza sativa L., which have been used for centuries in Chinese medicine. These herbs provide a comprehensive approach to treating respiratory conditions by addressing dryness, cough, and phlegm. Ethnopharmacological studies have explored the scientific basis of these herbs and identified active compounds that contribute to their medicinal effects. The traditional usage of MMDD by different ethnic groups reflects their knowledge and experiences. Examining this formula contributes to the understanding and development of ethnopharmacology.AIM OF THE STUDY:In the case of pulmonary fibrosis (PF), treating it can be challenging due to the limited treatment options available. This study aimed to assess the potential of MMDD as a treatment for PF by targeting macrophages and the PI3K/Akt/FOXO3a signaling pathway.MATERIALS AND METHODS:In a mouse model of PF, we investigated the effects of MMDD on inflammation, fibrosis, and M2 macrophage infiltration in lung tissue. Additionally, we examined the modulation of pro-fibrotic factors and key proteins in the PI3K/Akt/FOXO3a pathway. In vitro experiments involved inducing M2-type macrophages and assessing the impact of MMDD on fibroblast activation and the PI3K/Akt/FOXO3a pathway.RESULTS:Results demonstrated that MMDD improved weight, reduced inflammation, and inhibited M2 macrophage infiltration in mouse lung tissue. It downregulated pro-fibrotic factors, such as TGF-β1 and PDGF-RB, as well as markers of fibroblast activation. MMDD also exhibited regulatory effects on key proteins in the PI3K/Akt/FOXO3a signaling pathway.CONCLUSIONS:MMDD inhibited M2 macrophage polarization and released profibrotic factors that inhibited pulmonary fibrosis. As a result, the PI3K/Akt/FOXO3a signaling pathway is suppressed. MMDD is proving to be a successful treatment for PF. However, further research is needed to validate its effectiveness in clinical practice.

目的 探讨二补助育改良方对高龄小鼠子宫内膜形态学及氧化应激相关指标的影响.方法 将8～9月龄妊娠小鼠据随机数字表分为高龄模型组、补佳乐组(0.3 mg/kg)、阿司匹林组(15 mg/kg)和二补助育改良方低、中、高(10.89、21.78、43.56 g/kg)剂量组,另选择6～8周龄妊娠小鼠为育龄空白组,每组8只.观察子宫大体形态、子宫内膜形态、子宫内膜胞饮突、胚胎着床情况,计算各组胞饮突评分、妊娠率、平均着床位点数,WST-1法测定血清与子宫内膜中超氧化物歧化酶(SOD)活力,比色法测定丙二醇(MDA)的含量.结果 与育龄空白组比较,高龄模型组子宫呈黯棕红色,管腔内有瘀血,内膜变薄,血管、腺体数量减少,微绒毛稀疏,胞饮突评分极低(P<0.05),平均着床位点数减少(P<0.05),血清、子宫SOD活力降低(P<0.05),血清、子宫MDA含量升高(P<0.05).与高龄模型组比较,各治疗组子宫颜色、管腔内淤血、内膜厚度、血管与腺体数量均有不同程度改善,改良方低、中、高剂量组和补佳乐组胞饮突评分提高(P<0.05),改良方低、中、高剂量组平均着床位点数及妊娠率提高(P<0.05),改良方高剂量组与阿司匹林组血清SOD活力升高(P<0.05),改良方低、中、高剂量组与补佳乐组子宫SOD活力升高(P<0.05),改良方低、中、高剂量组和补佳乐组血清MDA含量降低(P<0.05),改良方中、高剂量组子宫MDA含量降低(P<0.05).结论 二补助育改良方可丰富内膜微绒毛和胞饮突的表达,提高SOD活力,降低MDA含量,改善血清与子宫氧化应激水平,从而改善子宫内膜容受性,提高胚胎着床位点数和妊娠率.

目的 探讨不同半夏炮制品体外抗肺纤维化的作用及筛选出的半夏制品对肺纤维化小鼠的干预作用.方法 用10μg/L转化生长因子β1(TGF-β1)诱导人胚肺成纤维细胞系(MRC-5)建立肺纤维化体外模型,经水提生半夏(2.1、4.2、8.4、16.7 g/L)、清半夏(2.1、4.2、8.4、16.7 g/L)、法半夏(2.1、4.2、8.4、16.7 g/L)和姜半夏(2.1、4.2、8.4、16.7 g/L)分别处理48 h后,通过CCK-8法检测细胞活性、天狼星红染色法测定胶原含量进行抗纤维化筛选.用Western blot分析筛选出的生半夏干预MRC-5细胞表达α-平滑肌肌动蛋白(α-SMA)的情况.C57BL/6N小鼠经口腔进行气管滴注博来霉素(2 mg/kg)建立肺纤维化模型,第14天开始灌胃生半夏水提液(2.5、5.0、10.0 g/kg),第28天取肺组织经HE和Masson染色观察,并测定羟脯氨酸(HYP)含量.结果 与对照组比较,TGF-β1诱导后的MRC-5细胞对生半夏(8.4 g/L)的细胞活性抑制作用更敏感(P<0.05),而未经TGF-β1诱导的MRC-5细胞则需要更高浓度的生半夏或清半夏(16.7 g/L)抑制细胞活性(P<0.05).与模型对照组比较,生半夏(8.4、16.7 g/L)能够抑制TGF-β1诱导的胶原合成(P<0.05),清半夏、法半夏、姜半夏各组未见抑制效应.与模型对照组比较,生半夏(2.1、4.2、8.4 g/L)能够抑制TGF-β1诱导的MRC-5细胞中 α-SMA蛋白的表达(P<0.05).生半夏(10 g/kg)能够缓解肺纤维化模型小鼠肺间质中的炎性浸润,恢复肺泡结构,减少胶原沉积,降低肺组织中的HYP含量(P<0.05).结论 生半夏对TGF-β1诱导后的病理性肌成纤维细胞具有活性抑制作用,并能阻止成纤维细胞向肌成纤维细胞分化,抑制其胶原合成,缓解模型小鼠的肺纤维化进展,为临床应用半夏治疗肺纤维化提供新的思路.

目的 探讨二补助育改良方对高龄小鼠子宫内膜胞饮突、微绒毛、雌激素受体α(estrogen receptorα,ERα)、白血病抑制因子(leukemia inhibitory factor,LIF)的影响.方法 将8～9月龄妊娠小鼠按照随机数字表分为高龄模型组,二补助育改良方低(10.89 g/kg)、中(21.78 g/kg)、高剂量组(43.56 g/kg),补佳乐组(0.3 mg/kg),阿司匹林组(15 mg/kg),另选择6～8周龄妊娠小鼠为育龄空白组,每组8只.育龄空白组和高龄模型组小鼠每日灌服0.5％羧甲基纤维素钠(CMC)溶液,其余各组小鼠每日灌服相应药物,连续灌胃5d.通过扫描电镜观察各组小鼠子宫内膜胞饮突、微绒毛表达,免疫组化法对子宫内膜ERα、LIF进行蛋白定位,Western blot和RT-qPCR法检测子宫内膜ERα、LIF蛋白及基因的表达量.结果 扫描电镜下,育龄空白组子宫内膜表面形态平整,微绒毛短而致密,胞饮突表达丰富;高龄模型组子宫内膜表面凹凸不平、破坏严重,微绒毛稀疏,几乎不可见胞饮突;各给药组微绒毛及胞饮突表达均优于高龄模型组.与育龄空白组比较,高龄模型组子宫内膜ERαmRNA及蛋白表达水平升高(P<0.05),LIF mRNA及蛋白表达水平均降低(P<0.05);与高龄模型组比较,各给药组子宫内膜ERαmRNA及蛋白表达水平均降低(P<0.05),LIF mRNA及蛋白表达水平均升高(P<0.05).结论 二补助育改良方可丰富子宫内膜微绒毛和胞饮突的表达,降低ERαmRNA及蛋白表达,提高LIF mRNA及蛋白表达,从而改善子宫内膜容受性.

BACKGROUND:We investigated the effect of Erbu Zhuyu decoction (EBZY) on angiogenesis via uterine natural killer (uNK) cells and the PI3K/Akt/eNOS pathway in embryo implantation dysfunction (EID) mice. METHODS:Pregnant mice were randomly divided into blank, model, EBZY, progynova, and aspirin groups. Uteri were excised on the 5th day of pregnancy for analysis. RESULTS:Mice in the model group showed pale uteri, a reduced implantation rate, and lower expression levels of phosphatidylinositol-3-kinase (PI3K), protein kinase B (Akt), endothelial nitric oxide synthase (eNOS) and nitric oxide (NO). Compared to the model group, implantation rates in the medium-dose and high-dose groups of EBZY were significantly higher (P < .05), PI3K and Akt mRNA expression levels in the low-dose group were significantly higher (P < .05, P < .01), and the expression of p-PI3K, p-Akt, and p-eNOS proteins in all treatment groups were significantly increased (P < .01, P < .05). The expression of NO was significantly increased in the low-dose and high-dose groups (P < .01, P < .05, respectively). The level of p-Akt protein in the high-dose group was significantly higher than those in the other treatment groups (P < .01, P < .05). There was no significant difference in the density of uNK cells (P > .05). CONCLUSIONS:EBZY facilitated embryo implantation in EID mice by enhancing endometrial angiogenesis via activation of the PI3K/Akt/eNOS pathway, at least in part. There was no evidence to indicate that EBZY could adjust the expression of uNK.

目的:探讨二补助育汤对胚胎着床障碍模型小鼠子宫内膜形态及血管生成素-1(Ang-1)mRNA、血管内皮生长因子(VEGF)mRNA的表达和定位的影响.方法:24只ICR雌性小鼠随机分为空白组、模型组、戊酸雌二醇组、二补助育汤组,每组6只,用米非司酮建立胚胎着床障碍动物模型,各组给予相应药物灌胃,妊娠第5天处死小鼠后,检测各组妊娠率、平均着床位点数、子宫内膜Ang-1和VEGF mRNA表达量及其蛋白定位.结果:模型组小鼠平均胚胎着床位点数、Ang-1 mR-NA、VEGF mRNA表达量明显低于空白组(均P<0.05);与模型组比较,二补助育汤组平均胚胎着床位点数、Ang-1 mRNA、VEGF mRNA表达量显著提高(均P<0.05).结论:二补助育汤可提高子宫内膜Ang-1和VEGF蛋白表达量,促进子宫内膜血管生成,从而提高子宫内膜容受性.

目的 观察补肾活血通络法治疗薄型子宫内膜肾虚血瘀证的临床疗效.方法 选取68例薄型子宫内膜患者,采用随机数字表法将入组患者随机分为治疗组34例、对照组34例.剔除脱落病例后,治疗组30例、对照组30例.治疗组予补肾活血通络方加减治疗,对照组予口服戊酸雌二醇加黄体酮胶囊治疗.观察子宫内膜厚度、子宫内膜类型、子宫内膜血流类型、子宫内膜血流阻力指数(RI)和搏动指数(PI)值、子宫动脉RI和PI值、中医症状评分.结果 治疗后2组子宫内膜厚度均较治疗前增厚,差异均有统计学意义(P＜0.05).治疗后治疗组A型内膜比例高于对照组、C型内膜比例低于对照组,差异均有统计学意义(P＜0.05),治疗组优于对照组.治疗后2组子宫内膜血流类型均较治疗前改善(P＜0.05),治疗组优于对照组,差异均有统计学意义(P＜0.05).治疗后,治疗组患者子宫内膜血流RI,PI较治疗前降低(P＜0.05),治疗后2组患者子宫动脉血流RI,PI较治疗前降低(P＜0.05).治疗后2组月经量、行经时间评分较本组治疗前均降低,差异均有统计学意义(P＜0.05).治疗后,治疗组月经色质、腰膝酸软、头晕耳鸣、行经腹痛评分较本组治疗前均降低,差异均有统计学意义(P＜0.05).治疗后,治疗组各症状及总积分较对照组降低明显,治疗前后症状评分差值比较,差异均有统计学意义(P＜0.05).结论 补肾活血通络法治疗薄型子宫内膜肾虚血瘀证具有较好的临床效果.

目的:探讨二补助育汤对着床障碍小鼠内膜中环氧化酶-2(COX-2)、过氧化物酶体增殖物激活受体δ(PPARδ)表达的影响.方法:将C57妊娠小鼠随机分为空白、模型、阿司匹林、戊酸雌二醇片、二补助育汤高、中、低剂量组,每组8只,吲哚美辛油制备胚胎着床障碍小鼠模型,各组给予相应药物灌胃.妊娠第5天取小鼠子宫,观察外观形态、COX-2、PPARδ基因和蛋白表达.结果:模型组小鼠子宫苍白细长,串珠样改变少,且分布稀疏不均.各观察组小鼠子宫较模型组红润,有串珠状改变,串珠分布增多且不均.模型组COX-2和PPARδ蛋白表达低于空白组(P＜0.05);各观察组COX-2蛋白表达均高于模型组(均P＜0.05).二补助育汤低、高剂量组PPAR8 mRNA表达较模型组、戊酸雌二醇片组明显增加(P＜0.05),阿司匹林组PPAR8 mRNA表达高于二补助育汤中剂量组(P＜0.05);二补助育汤各剂量组PPARδ蛋白表达高于模型组(P＜0.01),其中中剂量组高于戊酸雌二醇片组和阿司匹林组(P＜0.05).结论:二补助育汤能够调节子宫内膜COX-2和PPARδ表达,提高子宫内膜容受性,有利于胚胎着床.

目的 探讨甘草酸对转化生长因子β1(TGF-β1)诱导的大鼠肾间质成纤维细胞(NRK-49 F)中纤维化相关因子表达的影响.方法 体外培养NRK-49 F细胞,不同浓度甘草酸干预细胞48 h后,CCK-8法检测其对细胞增殖的影响,确定对细胞活性无显著影响的作用浓度.以10μg/L TGF-β1刺激NRK-49 F细胞构建肾间质纤维化细胞模型,加入不同浓度甘草酸进行干预,免疫荧光法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(COL1)表达情况.实时荧光定量PCR、Western Blot法检测纤维化相关因子α-SMA、COL1、Ⅲ型胶原(COL3)mRNA及蛋白表达情况,以评价甘草酸对成纤维细胞活化及细胞外基质分泌的影响.结果 CCK-8结果显示甘草酸在100～600μmol/L浓度范围内对NRK-49 F细胞活性无影响.免疫荧光结果表明,加入甘草酸后可抑制 α-SMA表达及COL1分泌,随甘草酸浓度增加,抑制作用增强.Western Blot结果显示,与模型组相比,甘草酸干预可下调α-SMA、COL1、COL3的蛋白表达,其中甘草酸150μmol/L组COL1的蛋白表达水平降低(P<0.01),甘草酸200μmol/L组COL1、COL3表达均降低(P<0.01).RT-PCR结果表明,与模型组相比,甘草酸干预可下调α-SMA、COL1、COL3表达,甘草酸150μmol/L组COL1 mRNA的表达水平降低(P<0.01),甘草酸200μmol/L组COL1、COL3 mRNA表达均降低(P<0.01).结论 甘草酸能够有效抑制TGF-β1诱导的NRK-49 F细胞中α-SMA、COL1、COL3的mRNA及蛋白表达,其作用机制有待进一步探讨.

目的:探索围着床期小鼠子宫内膜自噬情况.方法:选择36只ICR妊娠小鼠随机分为空白组、胚胎着床障碍模型组,每组小鼠18只.模型组采用米非司酮制备,每组每日予以生理盐水灌胃,分别于妊娠第4、5、6天脱颈处死小鼠各6只.观察子宫形态,计算各组平均胚胎着床位点数,检测自噬标志物微管相关轻链蛋白3-Ⅱ(LC3-Ⅱ),透射电镜观察子宫内膜细胞中自噬小体的表达.结果:空白组妊娠第4天(pd4)子宫无明显串珠样改变,妊娠第5天(pd5)、妊娠第6天(pd6)子宫串珠样改变明显且分布均匀.模型组pd4、pd5未见明显串珠样改变,pd6可见少量串珠稀疏分布.空白组pd6的胚胎着床位点数较pd4增多,差异有统计学意义(P<0.05);而pd5的胚胎着床位点数与pd4比较有增多,但差异无统计学意义(P>0.05).模型组pd5、pd6平均胚胎着床位点数分别低于空白组pd5、pd6(P<0.05,P<0.01).2组LC3-Ⅱ蛋白水平差异无统计学意义(P>0.05).透射电镜下空白组小鼠子宫内膜细胞完整,pd5自噬小体数量较pd4减少,pd6又增多.模型组部分细胞损伤、坏死,pd4自噬小体表达较空白组多,亦较模型组pd5、pd6多.结论:自噬参与并调控胚胎着床过程,自噬异常可能破坏子宫内膜细胞活性和功能,影响胚胎着床.

目的 观察二补助育汤对胚胎着床障碍小鼠子宫内膜自噬作用的影响.方法 将24只ICR妊娠小鼠随机分为空白组、模型组、补佳乐组、二补助育汤组,每组小鼠6只.胚胎着床障碍小鼠模型采用米非司酮制备,每组予以相应药物灌胃,妊娠第5天脱颈处死.计算各组小鼠平均胚胎着床位点数,检测自噬标志物微管相关轻链蛋白3-Ⅱ(LC3-Ⅱ)、Beclin-1表达,透射电镜观察子宫内膜细胞中自噬小体的表达.结果 模型组平均着床位点数低于空白组(P<0.05),补佳乐、二补助育汤组高于模型组(P>0.05).各组小鼠子宫内膜细胞中均见自噬小体,空白组见少量自噬小体表达;模型组部分细胞有破坏、坏死,内膜发育不良,细胞内自噬小体多于其他组;补佳乐组和二补助育汤组子宫内膜腺体和血管较模型组丰富,细胞内自噬小体表达虽多于空白组,但较模型组少.模型组LC3-Ⅱ、Beclin-1蛋白水平均低于空白组,其中LC3-Ⅱ有显著差异(P<0.05);补佳乐组和二补助育汤组LC3-Ⅱ表达高于模型组(P>0.05);二补助育汤组Beclin-1表达高于模型组和补佳乐组(P<0.05).结论 二补助育汤能够通过调节子宫内膜自噬作用,维持细胞稳态,从而利于胚胎着床.