BACKGROUND:Gastric motility disorder is an increasingly common problem among people with diabetes. Neurotransmitters have been recognized as critical regulators in the process of gastric motility. Previous study has shown that herb pair huanglian-banxia (HL-BX) can improve gastric motility, but the underlying mechanism is still unclear. The aim of this study was to further investigate the role of HL-BX in modulating brain-gut neurotransmission to promote gastric motility in diabetic rats, and to explore its possible mechanism.METHODS:The diabetic rats were divided into five groups. Gastric emptying rate, intestinal propulsion rate, body weight, and average food intake were determined. Substance P (SP), 5- hydroxytryptamine (5-HT), and glucagon-like peptide -1 (GLP-1) in the serum were measured by enzyme-linked immunosorbent assay. Dopamine (DA) and norepinephrine (NE) in the brain were analyzed by high-pressure liquid chromatography with a fluorescence detector. Protein expression of the tissues in the stomach and brain was determined by Western blot.KEY RESULTS:HL-BX reduced average food intake significantly, increased body weight, and improved gastric emptying rate and intestinal propulsion rate. HL-BX administration caused a significant increase in SP, GLP-1, and 5-HT, but a significant decrease in DA and NE. Interestingly, HL-BX regulated simultaneously the different expressions of MAPK and its downstream p70S6K/S6 signaling pathway in the stomach and brain. Moreover, berberine exhibited a similar effect to HL-BX.CONCLUSIONS:These results indicated that HL-BX promoted gastric motility by regulating brain-gut neurotransmitters through the MAPK signaling pathway. HL-BX and MAPK provide a potential therapeutic option for the treatment of gastroparesis.

INTRODUCTION:The function of promoting bone regeneration of Moutan Cortex (MC), a traditional Chinese medicine, has been widely known but, the effective components of MC in promoting osteoblast-mediated bone regeneration were still unclear.OBJECTIVE:The method of osteoblast membrane bio-specific extraction conjugated with HPLC analysis was established to screen bone regeneration active components from MC.METHODS:The fingerprints, washing eluate and desorption eluate of MC extract were analyzed by the established HPLC-DAD method. The established MC3T3-E1 cells membrane chromatography method was used for the bio-specific extraction of MC. The isolated compounds were identified by MS spectrometry. The effects and possible mechanisms of the isolated compounds were evaluated by molecular docking, ALP activity, cell viability by MTT Assay and proteins expression by Western Blot Analysis.RESULTS:The active compound responsible for bone regeneration from MC was isolated using the established method of osteoblast membrane bio-specific extraction conjugated with HPLC analysis, and it was identified as 1,2,3,4,6-penta-O-β-galloyl-D-glucose (PGG) by MS spectrometry. It was further demonstrated through molecular docking that PGG could fit well into the functional ALP, BMP2, and Samd1 binding pocket. The proliferation of osteoblasts was promoted, the level of ALP was increased, and the protein expression of BMP2 and Smad1 was increased as shown by further pharmacological verification.CONCLUSION:It was concluded that PGG, the bone regeneration active compound from MC, could stimulate the proliferation of osteoblasts to promote osteoblast differentiation, and its mechanism might be related to the BMP/Smad1 pathway.

建立了3-脱氧果糖(3-DF)初步检测的高效液相色谱(HPLC)方法.先使用气相色谱―质谱联用(GC-MS)对合成的3-DF标样进行鉴定.再采用C18反相色谱柱,流动相为乙腈-水(10％:90％～90％:10％)梯度洗脱60 min,紫外检测波长352 nm,应用2,4-二硝基苯肼柱前衍生法,以薄层色谱―高效液相色谱联用(TLC-HPLC)对3-DF衍生物峰进行定位,并完善衍生条件,进行方法学考察.结果表明:合成样品为3-DF;衍生条件以衍生液中1:3添加乙酸乙酯效果最佳;3-DF衍生物峰出峰时间为15.972 min,呈尖峰,分离度良好,线性范围为0.5～8 mg/mL,方法精密度RSD为2.4％,重复性RSD为4.4％,稳定性RSD为2.4％.本检测方法稳定、可靠,可初步用于3-DF定性及相对定量.

结合近五年糖尿病胃轻瘫的基础实验研究,对中药干预糖尿病胃轻瘫的的胃肠动力信号通路研究进行综述.发现干细胞因子/酪氨酸激酶信号通路、Ras同源A蛋白/RhoA相关蛋白激酶RhoA/ROCK等多条信号通路参与了糖尿病胃轻瘫的发生和发展过程;中药治疗糖尿病胃轻瘫疗效确切,具有多角度多靶点优势,但缺乏物质基础研究,需要开展符合辨证论治和遣方用药规律的物质基础和作用机理研究,以期为临床有效治疗糖尿病胃轻瘫提供思路.

目的:研究半夏泻心汤对3-脱氧葡萄糖醛酮(3DG)诱导的大鼠骨骼肌胰岛素抵抗的干预作用及作用机制.方法:将SD大鼠随机分为6组:正常对照组、模型组、二甲双胍组(剂量为0.2 g/kg)和半夏泻心汤低、中、高剂量组(剂量分别为1 g/kg、3 g/kg、5 g/kg),每组10只.每天灌胃给药1次,共2周.给药2周后,进行葡萄糖耐量试验;留取血清,测定空腹血糖(FBG)、空腹胰岛素(FINS);提取大鼠骨骼肌膜蛋白和总蛋白,采用West?ern Blot法分别检测骨骼肌膜上的葡萄糖转运蛋白4(GLUT4)、胰岛素受体蛋白-β(IR-β)及其磷酸化蛋白表达量.结果:与正常对照组比较,模型组血糖、葡萄糖曲线下面积(AUC)、FBG、FINS水平明显升高(P<0.05),GLUT4、p-IR-β/IR-β水平明显降低(P<0.01);与模型组比较,半夏泻心汤各剂量组血糖、AUC、FBG、FINS水平明显降低(P<0.05),GLUT4、p-IR-β/IR-β水平明显升高(P<0.05或P<0.01).结论:半夏泻心汤可改善大鼠胰岛素抵抗,其机制可能是通过上调骨骼肌胰岛素信号通路GLUT4、p-IR-β蛋白表达,促进骨骼肌葡萄糖摄取,从而改善大鼠骨骼肌胰岛素抵抗.

Objective To explore the differences of intestinal flora in patients with type 2 diabetes mellitus（T2DM） with different TCM syndromes through the high-throughput 16S rRNA sequencing analysis.Methods According to TCM syndrome types, 200 inpatients with T2DM were divided into Yin deficiency and heat excess syndrome（40 cases）,damp-heat accumulating spleen syndrome（33 cases）,QI-YIN deficiency syndrome（103 cases） and Yin-Yang deficiency syndrome（24 cases）.At the same time, 26 healthy patients were also involved as the healthy control group.The clinical data, serum and fecal specimens were collected, and the fecal specimens were sequenced by 16s rRNA high-throughput sequencing technique.The software R3.4.1 was used to analyze the composition and diversity of intestinal flora.LEfSe method was used to compare intestinal flora markers differences and analyze the correlation of these markers and clinical data.The change characteristics of intestinal flora function were predicted by the PICRUSt.Results The ratio of Firmicutes to Bacteroidetes in each T2DM syndrome group was lower than that in the healthy control group.The intestinal flora diversity of T2DM patients was significantly different from that of the healthy control group（P＜0.05）,and that of the damp-heat accumulating spleen syndrome group was the lowest.Venn diagram analysis showed that 70 OTUs only appeared in the Qi-Yin deficiency syndrome group, while 13,4 and 8 OTUs in Yin deficiency and heat excess syndrome group, damp-heat accumulating spleen syndrome group and Yin-Yang deficiency syndrome group, respectively.MRPP analysis showed that the difference of intestinal flora structure in different syndromes of T2DM among groups was greater than that within groups（P＜0.05）.The significant difference analysis of species at the level of genus showed that there were 16 genera with statistical differences among the four groups.LEfSe analysis showed that among the four groups of T2DM,Yin-Yang deficiency syndrome group showed much more diversity, with 8 representative intestinal floras, followed by Qi-Yin deficiency syndrome and Yin deficiency and heat excess syndrome, each with 2 representative floras.The correlation analysis between representative floras and clinical data showed that there was correlation with disease course, body mcss index（BMI）,lipid metabolism and so on.The damp-heat accumulating spleen syndrome ran through the whole course of diabetes and overlapped with other syndromes, no representative flora was found in this group.The prediction of intestinal flora function also confirmed that the four syndromes were different in carbohydrate metabolism and amino acid metabolism（P＜0.05）.Conclusion There are differences in flora structure in patients with T2DM with different TCM syndrome types, which may affect the progression of diabetes and the occurrence, development and outcome of different TCM syndromes.

目的 研究半夏泻心汤对3-脱氧葡萄糖醛酮(3 DG)致糖尿病前期大鼠肠道菌群的影响.方法 大鼠连续灌胃3DG(50 mg·kg-1·d-1)2周制得糖尿病前期大鼠模型.将雄性SD大鼠随机分为3组:正常组(7只)、模型组(7只)和实验组(3只).造模后,实验组灌胃半夏泻心汤水煎液(5 mL·kg-1),正常组和模型组灌胃蒸馏水,3组均连续灌服2周.收集3组大鼠粪.以高通量测序方法分析肠道整体微生物及OTU(operational taxonomic units)组成;基于菌群Alpha多样性和β多样性分析半夏泻心汤对糖尿病前期大鼠肠道菌群结构的影响;基于菌群属水平下的丰度分析差异菌属.结果 在17个样本中,共获得455个OTU,其中丰度最高的微生物门为拟菌门和厚壁菌门.正常组、模型组和实验组的Alpha多样性指数的Sobs指数分别为308.00±2.16,284.00±2.16和304.00±1.00;这3组的Ace指数分别为327.59±22.81,332.77±25.07和329.03±8.79;这3组的Chao指数为329.65±24.49,335.02±29.81和331.45±12.36;这3组的Shannon指数分别为3.35±0.17,3.46±0.20和3.41±0.18;这3组的Simpson指数分别为8.00×10-2±0.01,7.00×10-2±0.02和7.00×10-2±0.01.与正常组相比,模型组新增Enteractinococcus和Ignavigranum 2个菌属,且Facklamia、Papillibacter、Peptococcus和Parasutterella等4个菌属丰度显著上调,而Alistipes、Candidatus_Soleaferrea、Corynebacterium、Enterococcus、Tyzzerella和Faecalitalea等6个菌属丰度显著下调,差异均有统计学意义(P<0.05,P<0.01).与模型组相比,实验组Faecalitalea、Parasutterella以及Tyzzerella菌属丰度显著上调,而Corynebacterium和Facklamia菌属丰度显著下调,差异均有统计学意义(P<0.05,P<0.01).结论 半夏泻心汤对糖尿病前期的治疗作用与其对肠道菌群的调控有关.

Type 2 diabetes (T2D) is a major global public health burden, with β-cell dysfunction a key component in its pathogenesis. However, the exact pathogenesis of β-cell dysfunction in T2D is yet to be fully elucidated. Ferroptosis, a recently discovered regulated form of non-apoptotic cell death, plays a vital role in the development of diabetes and its complications. The current study aimed to identify the key molecules involved in β-cell ferroptosis3 in patients with T2D using the mRNA expression profile data of GSE25724 by bioinformatic approaches. The differentially expressed mRNAs (DE-mRNAs) in human islets of patients with T2D were screened using the islet mRNA expression profiling data from the Gene Expression Omnibus and their intersection with ferroptosis genes was then obtained. Ferroptosis-related DE-mRNA functional and pathway enrichment analysis in T2D islet were performed. Using a protein-protein interaction (PPI) network constructed from the STRING database, Cytoscape software identified ferroptosis-related hub genes in the T2D islet with a Degree algorithm. We constructed a miRNA-hub gene network using the miRWalk database. We generated a rat model of T2D to assess the expression of hub genes. A total of 1,316 DE-mRNAs were identified in the islet of patients between T2D and non-T2D (NT2D), including 221 and 1,095 up- and down-regulated genes. Gene set enrichment analysis revealed that the ferroptosis-related gene set was significantly different in islets between T2D and NT2D at an overall level. A total of 33 ferroptosis-related DE-mRNAs were identified, most of which were significantly enriched in pathways including ferroptosis. The established PPI network with ferroptosis-related DE-mRNAs identified five hub genes (JUN, NFE2L2, ATG5, KRAS, and HSPA5), and the area under the ROC curve of these five hub genes was 0.929 in the Logistic regression model. We constructed a regulatory network of hub genes and miRNAs, and the results showed that suggesting that hsa-miR-6855-5p, hsa-miR-9985, and hsa-miR-584-5p could regulate most hub genes. In rat model of T2D, the protein expression levels of JUN and NFE2L2 in pancreatic tissues were upregulated and downregulated, respectively. These results contribute to further elucidation of ferroptosis-related molecular mechanisms in the pathogenesis of β-cell dysfunction of T2D.

目的 探索半夏泻心汤对3-脱氧葡萄糖醛酮(3 DG)诱导的糖尿病前期大鼠的保护作用及机制.方法 SD大鼠连续灌服50 mg·kg-13DG,14 d,建立糖尿病前期大鼠模型.将造模成功大鼠按体质量随机分为模型组和实验组,每组10只;另取10只正常大鼠作为正常组.正常组和模型组均按10 mL·kg-1·d-1剂量灌胃给予生理盐水;实验组灌胃给予相当于5 g·kg-1·d-1生药的半夏泻心汤,每组均连续灌服14 d.采用酶联免疫吸附法测定结肠和血清中蛋白质羰基化程度、活性氧(ROS)以及白细胞介素-1β(IL-1β)含量.结果 正常组、模型组和实验组的结肠蛋白质羰基化程度分别为(3.20±0.21),(4.31±0.25)和(3.72±0.23)nmol·mg-1;这3组结肠ROS表达量分别为(738.10±17.29),(1082.00±51.93)和(834.80±27.63)U·mg-1;这3组结肠IL-1β表达量分别为(176.10±9.95),(237.70±16.08)和(200.10±11.59)ng·L-1.这3组血清中蛋白质羰基化程度分别为(0.53±0.06),(0.73±0.06)和(0.61±0.06)nmol·mg-1;这3组血清中ROS表达量分别为(295.60±38.90),(446.70±39.86)和(331.40±10.13)U·mL-1;这3组血清中IL-1β含量分别为(25.33±0.90),(29.83±2.44)和(25.93±1.02)ng·L-1.模型组的上述指标与正常组相比,实验组的上述指标与模型组相比,差异均有统计学意义(均P<0.05).结论 半夏泻心汤对糖尿病前期的保护作用可能与其降低结肠和血清中蛋白质羰基化水平、ROS水平以及炎症因子含量有关.

目的 探讨半夏泻心汤剂对3-脱氧葡萄糖醛酮(3-deoxyglucosone,3DG)诱导的糖尿病前期(impaired glucose regulation,IGR)大鼠血糖、胰岛素、胰高血糖素样肽-1(glucagon-like peptide-1,GLP-1)以及炎症因子的影响.方法 SD大鼠上午灌服50 mg·kg-1 3DG,连续灌服14 d,建立IGR大鼠模型.将造模成功大鼠按体质量随机分为模型组和实验组,每组12只;另取12只正常大鼠作为空白组.空白组和模型组均按10 mL·kg-1·d-1容积灌胃给予0.9％NaCl;实验组下午伴随灌服生药5 g·kg-1·d-1,连续14d.干预14 d后,进行空腹血糖、口服糖耐量试验(oral glucose tolerance test,OGTT),用酶联免疫吸附实验法检测OGTT 30 min血清胰岛素和血浆GLP-1水平.结果 实验组、模型组和空白组的空腹血糖分别为(4.85±0.26),(5.35±0.38)和(4.59±0.38)mmol·L-1,30 min 血糖分别为(8.42±0.48),(10.21±0.45)和(7.65±0.43)mmol·L-1,60 min 血糖分别为(7.51±0.46),(8.72±0.46)和(6.89±0.42)mmol·L-1,血清胰岛素分别为(13.76±1.45),(11.34±1.11)和(13.89±1.37)mU·L-1,血浆 GLP-1 分别为(3.12±0.25),(2.68±0.17)和(3.36±0.31)pmol·L-1;模型组的上述指标与实验组和空白组比较,差异均有统计学意义(均P＜0.05).结论 半夏泻心汤剂可改善3DG所致的大鼠糖调节受损,这可能与其增加GLP-1水平和降低炎症水平有关.

Ethnopharmacological relevance: Atopic dermatitis (AD) is a complex skin disease with highly heterogeneous inflammation, which ranks among the largest component of the nonfatal diseases worldwide. The medications currently used to treat AD primarily include antihistamines, vitamin D and anti-inflammatory drugs, etc. But, the usage of these drugs is usually accompanied by various side-effects. Formononetin (FMN), a natural active ingredient of Astragalus membranaceus (Fisch.) Bunge, decreases the AD relapse rate, reduces recurring severity incidence and resists the inflammation in the initial stage of AD. However, the underlying mechanism of FMN on repressing the development of AD is still unknown. Aim of the study: To investigate the potential mechanism of FMN on relieving the initial responses of AD and elucidate its possible therapeutic targets in vivo and in vitro. Materials and methods: A fluorescein isothiocyanate (FITC)-induced mouse model of the initial stage of AD was established in vivo. Human keratinocytes (HaCaT) cells were co-stimulated with tumor necrosis factor alpha (TNF-alpha) and polyinosinic-polycytidylic acid (Poly(I:C)) in vitro. The production of thymic stromal lymphopoietin (TSLP) and immunoglobulin E (IgE) were detected by enzyme-linked immunosorbnent assay (ELISA). The protein expression was measured through immunohistochemistry and western blotting. The mRNA expression was examined by real-time quantitative polymerase chain reaction (RT-qPCR). The impact of TNF-alpha-induced protein 3 (TNFAIP3/A20) was reflected using its small interfering RNA (siRNA). The role of G protein-coupled estrogen receptor (GPER) was explored using its agonist (G1), antagonist (G15) or siRNA (siGPER) in vitro. Results: We found that FMN upregulated the expression of A20 protein and mRNA in the initial stage of AD model, especially in the epithelial region of ear tissue, and inhibited the production of TSLP simultaneously. Consistently, FMN significantly upregulated A20 protein and its mRNA expression while reduced TSLP protein and its mRNA expression in vitro, and this effect could be antagonized by A20 siRNA (siA20). Moreover, compared with PPT (ER alpha agonist) and DPN (ER beta agonist), G1 could significantly increase the expression of A20. In addition, compared with MPP (ER alpha antagonist) and PHTPP (ER beta antagonist), G15 could markedly reduce the expression of A20. Furthermore, the effects of FMN on A20 were interfered by siGPER and G15 in vitro and in vivo. Conclusions: These results demonstrated that FMN attenuated AD by upregulating A20 expression via activation of GPER. This new strategy might have effective therapeutic potential for AD and other inflammatory disorders.

Aconitine, the main component in Radix Aconiti Lateralis Preparata, not only exerts the anti-tumor effect on Hepatocellular Carcinoma (HCC) but also damages on immune system. In the present study, Crude Monkshood Polysaccharide (CMP), another one natural composition component originated from the same herbal with aconitine, combined with aconitine to investigate the effects on HCC and immunity in vitro and in vivo. The combination of CMP and aconitine enhanced the ability of the immunocyte to kill the tumor cell in vitro and had an additive effect on anti-HCC in vivo. Aconitine-CMP in combination improved the spleen weights, spleen index, thymus weights, thymus index. Elevated CD4+ T and CD8+ T cells and macrophages in spleen, decreased serum IL-6 level and increased serum IFN-γ and TNF-α levels were observed in mice treated with the combination of aconitine and CMP compare with control group (P<0.05). Our results showed that the combination of aconitine and CMP exerts anti-tumor effect by directly killing tumor cells and enhancing the anti-tumor immune responses, which further implies that chemotherapy drugs combined with Chinese medicine immunopotentiator maybe a feasible and effective strategy for HCC.

胰高血糖素样肽-1(GLP-1)可以刺激胰岛素释放以及抑制胰高血糖素释放,与2型糖尿病有关.苦味中药可以激活肠道上的苦味受体(TAS2Rs),通过磷脂酶C通路和磷酸二酯酶通路刺激肠道内分泌L细胞未分泌GLP-1,广泛用于2型糖尿病的防治.本文综述了苦味中药对肠道TAS2 Rs介导的GLP-1分泌的调控作用,为苦味中药防治2型糖尿病提供新视角.

目的:通过寒冷致小鼠上呼吸道黏膜免疫功能低下(upper airway immunity dysfunction,UAID)模型的实验,基于免疫球蛋白A单克隆抗体(immunoglobulin A monoclonal antibody,IgA MAb)阻断新型冠状病毒(SARS-CoV-2)的Spike-ACE 2相互作用探究RMK-03TM喷剂对上呼吸道黏膜的免疫调节机制.方法:将40只SPF级ICR小鼠分为正常组、UAID模型组、利巴韦林喷雾剂组、RMK-03TM喷雾剂组4组,每组10只;利巴韦林喷剂组和RMK-03TM喷剂组先给予口腔喷雾0.1 mL/只q12h,给药3 d,;第3 d,除正常组外其他各组小鼠复制UAID模型,即将小鼠放入-20℃寒冷环境中15 min,之后在室温适应1 h,经腹腔注射毛果芸香碱滴眼液(0.25 mL/只),2 min后以安乐处死小鼠,取口腔、咽喉部黏膜组织进行HE病理学切片检查,以Western blotting法考察其IgA、pIgR蛋白的表达,采用ELISA法检测细胞因子IL-4、IL-6和IFN-y、TNF-a含量;采用免疫组化法检测ACE 2和ACE的表达情况.结果:UAID模型组小鼠唾液中SIgA含量显著低于正常组(P＜0.05),提示模型复制成功;与正常组相比,U AID模型组小鼠的口腔和咽喉黏膜组织可见轻度间质水肿、小血管扩张充血,中性粒细胞伴有淋巴管轻度扩张水肿;UAID模型组小鼠口腔、咽喉部黏膜组织IgA、pIgR蛋白表达,IgA相关细胞因子IL-4、IL-6的含量及pIgR相关细胞因子IFN-γ、TNF-a的含量均显著低于正常组(P＜0.05),该组的ACE、ACE 2平均光密度值(IOD/area)均显著性高于正常组(P＜0.05);利巴韦林喷剂组和RMK-03TM喷剂组的上述主要指标均在不同程度上优于UAID模型组,且后者的上述主要指标优于前者.结论:RMK-03TM皮肤黏膜护理喷剂通过调节局部黏膜免疫屏障低下相关IgA的含量,抑制呼吸道黏膜中ACE 2活性表达,间接推测其阻断SARS-CoV-2的Spike-ACE 2相互作用以提供黏膜免疫.

Diabetic gastroparesis (DGP) is a serious and chronic complication of long-standing diabetes mellitus, which brings a heavy burden to individuals and society. Traditional Chinese medicine (TCM) is considered a complementary and alternative therapy for DGP patients. Huanglian (Coptidis Rhizoma, HL) and Banxia (Pinelliae Rhizoma, BX) combined as herb pair have been frequently used in TCM prescriptions, which can effectively treat DGP in China. In this article, a practical application of TCM network pharmacological approach was used for the research on herb pair HL-BX in the treatment of DGP. Firstly, twenty-seven potential active components of HL-BX were screened from the TCMSP database, and their potential targets were also retrieved. Then, the compound-target network and PPI network were constructed from predicted common targets, and several key targets were found based on the degree of the network. Next, GO and KEGG enrichment analyses were conducted to obtain several significantly enriched terms. Finally, the experimental verification was made. The results demonstrated that network pharmacological approach was a powerful means for identifying bioactive ingredients and mechanisms of action for TCM. Network pharmacology provided an effective strategy for TCM modern research.

Our previous research suggests that 3-deoxyglucosone (3DG), formed in the caramelization course and Maillard reactions in food, is an independent factor for the development of prediabetes. Since the relationship between type 2 diabetes (T2D) and intestinal microbiota is moving from correlation to causality, we investigated the alterations in the composition and function of the intestinal microbiota in 3DG-induced prediabetic rats. Rats were given 50 mg/kg 3DG by intragastric administration for two weeks. Microbial profiling in faeces samples was determined through the 16S rRNA gene sequence. The glucagon-like peptide 2 (GLP-2) and lipopolysaccharide (LPS) levels in plasma and intestinal tissues were measured by ELISA and Limulus test, respectively. 3DG treatment did not significantly change the richness and evenness but affected the composition of intestinal microbiota. At the phylum level, 3DG treatment increased the abundance of nondominant bacteria Proteobacteria but did not cause the change of the dominant bacteria. Meanwhile, the abundance of the Prevotellaceae family and Parasutterela genus and the Alcaligencaeae family and Burkholderiales order and its attachment to the Betaproteobacteria class were overrepresented in the 3DG group. The bacteria of Candidatus Soleaferrea genus, Gelria genus, and Thermoanaerobacteraceae family and its attachment to Thermoanaerobacterales order were apparently more abundant in the control group. In addition, 45 KEGG pathways were altered after two-week intragastric administration of 3DG. Among these KEGG pathways, 13 KEGG pathways were involved in host metabolic function related to amino acid metabolism, carbohydrate metabolism, metabolism of cofactors and vitamins, and metabolism of terpenoids and polyketides. Moreover, the increased LPS levels and the decreased GLP-2 concentration in plasma and intestinal tissues were observed in 3DG-treated rats, together with the impaired fasting glucose and oral glucose tolerance. The alterations in composition and function of the intestinal microbiota were observed in 3DG-treated rats, which provides a possible mechanism linking exogenous 3DG intake to the development of prediabetes.

The tumor immunological microenvironment in hepatocellular carcinoma (HCC) is the T-helper (Th) 2 dominant inhibition state. Improving the immunosuppressive tumor microenvironment represents an important strategy for HCC treatment. TSLP-OX40L pathway is a target to improve Th2 immunosuppression. Yupingfeng granule (YPF) is clinically used to effectively improve the immune status of HCC. In this study, YPF increased the percentage of mature dendritic cells (DCs) and decreased levels of TSLP, TSLPR, and OX40L in tumor and adjacent tissues of the orthotopic-HCC mice model. This occurs together with the decreased levels of Th2 cytokines and increased levels of Th1 cytokines and Th1/Th2 ratio. In vitro experiment showed that YPF not only increased the percentage of mature DCs and stimulated IL-12 secretion in DCs but also reduced the positive rate of OX40L expression, decreased the proportion of CD4(+) IL-13(+) T cells, increased levels of Th1 cytokines, and decreased levels of Th2 cytokines from TSLP-treated DCs. In summary, these findings demonstrated that YPF promoted the maturation of DCs, decreased OX40L in TSLP-induced DCs, and improved the immunosuppressive state of Th2 in HCC microenvironment. Our results suggest that the mechanism underlying the improving effect of YPF on the immunosuppression is related to the DC-mediated TSLP-OX40L pathway.

目的 研究麻黄-甘草药对对过敏性哮喘的影响,并初步探讨其作用机制.方法 建立屋尘螨(HDM)诱导的过敏性哮喘模型,造模同时给予麻黄-甘草药对提取物(MG).使用无创肺功能仪检测小鼠气道反应性,HE染色观察小鼠肺组织病理学变化,Masson染色观察小鼠肺组织的胶原沉积情况,进行血液嗜酸性粒细胞(EOS)计数,ELISA法检测支气管肺泡灌洗液(BALF)及肺组织IL-4、IL-5、IL-13、TGF-β1及血清IgE的表达,免疫组化检测肺组织E-cadherin、N-cadherin、Vimentin、α-SMA和TGF-β1蛋白表达情况.结果 MG可显著降低气道高反应性,抑制BALF及肺组织IL-4、IL-5和IL-13的表达,降低血液EOS及血清IgE的表达,并且减轻肺组织的炎症浸润及胶原沉积.进一步结果表明MG可抑制肺组织TGF-β1的表达,上调E-cadherin以及下调N-cadherin、Vimentin、α-SMA的表达.结论 MG能够通过抑制支气管上皮间质转化,从而缓解气道的炎症状态以及改善哮喘的气道重塑,进而发挥对过敏性哮喘的治疗作用.

Allergic asthma is a common chronic lung inflammatory disease and seriously influences public health. We aim to investigate the effects of formononetin (FMN) and calycosin (CAL), 2 flavonoids in Radix Astragali, on allergic asthma and elucidate possible therapeutic targets. A house dust mite (HDM)-induced allergic asthma mouse model and TNF-α and Poly(I:C) co-stimulated human bronchial epithelial cell line (16HBE) were performed respectively in vivo and in vitro. The role of G protein-coupled estrogen receptor (GPER) was explored by its agonist, antagonist, or GPER small interfering RNA (siGPER). E-cadherin, occludin, and GPER were detected by western blotting, immunohistochemistry, or immunofluorescence. The epithelial barrier integrity was assessed by trans-epithelial electric resistance (TEER). Cytokines were examined by enzyme-linked immunosorbent assay (ELISA). The results showed that flavonoids attenuated pulmonary inflammation and hyperresponsiveness in asthmatic mice. These flavonoids significantly inhibited thymic stromal lymphopoietin (TSLP), increased occludin and restored E-cadherin in vivo and in vitro. The effects of flavonoids on occludin and TSLP were not interfered by ICI182780 (estrogen receptor antagonist), while blocked by G15 (GPER antagonist). Furthermore, compared with PPT (ERα agonist) and DPN (ERβ agonist), G1 (GPER agonist) significantly inhibited TSLP, up-regulated occludin, and restored E-cadherin. siGPER and TEER assays suggested that GPER was pivotal for the flavonoids on the epithelial barrier integrity. Finally, G1 attenuated allergic lung inflammation, which could be abolished by G15. Our data demonstrated that 2 flavonoids in Radix Astragali could alleviate allergic asthma by protecting epithelial integrity via regulating GPER, and activating GPER might be a possible therapeutic strategy against allergic inflammation.

Glucose triggers glucagon-like peptide (GLP)-1 secretion from L cells involving several glucose sensors including sodium-glucose transporter (SGLT)1, glucose transporter (GLUT)2, and sweet taste receptors (STRs). This study investigated the effects of different glucose concentrations on GLP-1 secretion, intracellular concentrations of Ca2+ and cAMP, glucose uptake, and protein levels of SGLT1, GLUT2, and STRs in STC-1 cells. Low glucose (5.6 mM) increased GLP-1 secretion, intracellular Ca2+ concentration, and SGLT1 protein level compared with glucose-free group. GLP-1 secretion and intracellular Ca2+ concentration triggered by low glucose were inhibited by the SGLT1 inhibitor. GLP-1 secretion or intracellular Ca2+ concentration in high-glucose (25, 100, 200 mM) groups was significantly higher than that of low-glucose group. Elevation of cAMP level was observed in concentration-dependent manner, and decreased glucose uptake was observed in 100 or 200 mM glucose group. High glucose increased protein levels of STRs and GLUT2 in comparison to low-glucose group. GLP-1 secretion and intracellular levels of Ca2+ and cAMP triggered by high glucose were inhibited in the presence of the GLUT2 or STR inhibitor. These results suggest that SGLT1 is dominantly responsible for GLP-1 secretion triggered by low glucose, and that STRs and GLUT2 are involved in GLP-1 secretion induced by high glucose.