Chronic obstructive pulmonary disease (COPD) is a progressive lung disorder linked to oxidative stress, mitochondrial damage, and impaired mitophagy. Tanreqing (TRQ) inhalation solution is widely used for respiratory diseases but its mechanism against COPD remains poorly defined. Here we investigated the protective effects of TRQ and its underlying pathway in a COPD rat model and cigarette smoke extract (CSE)-stimulated A549 cells. Our results showed that TRQ treatment alleviated body weight loss, lung pathological injury, alveolar destruction, and airway remodeling in rats. TRQ reduced ROS and MDA levels, restored SOD activity, and improved mitochondrial function including ATP production, mitochondrial membrane potential, and mitochondrial permeability transition pore homeostasis. Moreover, TRQ enhanced autophagy and mitophagy by upregulating Bnip3, Nix, Pink1, and Parkin expression. In vitro, knockdown of Pink1 abolished TRQ-mediated mitophagy activation, confirming that the Pink1/Parkin pathway was essential for TRQ action. In conclusion, TRQ inhalation solution ameliorates COPD by suppressing oxidative stress and restoring mitochondrial homeostasis via activating Pink1/Parkin-mediated mitophagy. This study identifies TRQ as a promising candidate for COPD treatment.

ObjectiveTo establish a mouse model of hypoxic pulmonary hypertension (HPH) through intermittent hypoxia induction, and to develop a comprehensive and reliable evaluation system for the HPH model, thereby providing experimental evidence for mechanistic studies and translational applications related to this disease.MethodsTwenty-four male specific pathogen-free (SPF) BALB/c mice were randomly divided into a control group and a model group, with 12 mice per group. Mice in the model group were placed in a hypobaric oxygen chamber control system to establish the HPH model, whereas mice in the control group received no intervention. After 28 days of modeling, a comprehensive evaluation of the pathophysiological characteristics of both groups was performed using a general condition scoring scale, echocardiography, hemodynamic measurements, blood gas analysis, hematological tests, organ coefficient determination, histopathological examination, and enzyme-linked immunosorbent assay (ELISA). In addition, correlation analyses were conducted among echocardiographic parameters, the contents of endothelin‑1 (ET‑1) and N‑terminal pro‑B‑type natriuretic peptide (NT‑proBNP) in lung tissue, and other measured indices.ResultsWith prolonged modeling duration time, body weight, water intake, and activity level of mice in the model group were significantly decreased compared with the control group (P<0.05). Echocardiography showed that, compared with the control group, the model group exhibited increased right ventricular dimensions (P<0.01), right ventricular anterior wall thickening (P<0.01), and widened main pulmonary artery diameter (P<0.01), whereas the peak systolic velocity across the pulmonary valve, tricuspid annular plane systolic excursion, and peak systolic velocity of the tricuspid annulus were significantly decreased (P<0.001); tricuspid regurgitation was observed in some model animals. Hemodynamic results revealed that right ventricular systolic pressure was elevated in the model group compared with the control group (P<0.001). Blood gas analysis showed that pH, partial pressure of oxygen, oxygen saturation, actual bicarbonate, and total carbon dioxide were all decreased in the model group compared with the control group (P<0.01). Hematological indices demonstrated that lymphocyte counts (P<0.05) and reticulocyte counts (P<0.001) were decreased in the model group compared with the control group. Compared with the control group, the organ coefficients of heart and lung in the model group were significantly increased (P<0.05 and P<0.001). Pathological examination revealed that the right ventricular hypertrophy index was significantly increased in the model group (P<0.001), with varying degrees of damage to the right ventricle, pulmonary artery, and pulmonary vessels; both the pulmonary artery wall thickness percentage and the pulmonary wall area percentage were significantly elevated (P<0.001). ELISA results showed that the levels of ET‑1 and NT‑proBNP in lung tissues were significantly increased in the model group compared with the control group (P<0.05 and P<0.001, respectively). Correlation analysis indicated that some echocardiographic parameters were highly correlated with multiple indices in the development of HPH (P<0.05).ConclusionEchocardiography can accurately assess a series of hemodynamic changes in HPH, including right ventricular structural, functional impairment, and pulmonary hypertension. Laboratory tests not only help verify whether the model has been successfully established, but also provide deeper insights into the pathogenesis of HPH, evaluate the effects of interventions, and offer scientific evidence for clinical outcomes. Pathological examination can further confirm the alterations in pulmonary vascular remodeling and increased right heart load. This multimodal analysis provides a reliable animal model and evaluation paradigm for both basic and translational research on HPH, and is of great significance for exploring disease mechanisms and developing precision therapeutic strategies.

This study aimed to explore the pharmacological mechanism of Yourenji Capsules(YRJ) in improving osteoporosis by combining network pharmacology and proteomics technologies. The SD rats were randomly divided into a blank control group and a 700 mg·kg~(-1) YRJ group. The rats were subjected to gavage administration with the corresponding drugs, and the blank serum, drug-containing serum, and YRJ samples were compared using ultra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) to analyze the main components absorbed into blood. Network pharmacology analysis was conducted based on the YRJ components absorbed into blood to obtain related targets of the components and target genes involved in osteoporosis, and Venn diagrams were used to identify the intersection of drug action targets and disease targets. The STRING database was used for protein-protein interaction(PPI) network analysis of potential target proteins to construct a PPI network. Gene Ontology(GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment were performed using Enrichr to investigate the potential mechanism of action of YRJ. Ovariectomy(OVX) was performed to establish a rat model of osteoporosis, and the rats were divided into a sham group, a model group, and a 700 mg·kg~(-1) YRJ group. The rats were given the corresponding drugs by gavage. The femurs of the rats were subjected to label-free proteomics analysis to detect differentially expressed proteins, and GO functional enrichment and KEGG pathway enrichment analyses were performed on the differentially expressed proteins. With the help of network pharmacology and proteomics results, the mechanism by which YRJ improves osteoporosis was predicted. The analysis of the YRJ components absorbed into blood revealed 23 bioactive components of YRJ, and network pharmacology results indicated that key targets involved include tumor necrosis factor(TNF), tumor protein p53(TP53), protein kinase(AKT1), and matrix metalloproteinase 9(MMP9). These targets are mainly involved in osteoclast differentiation, estrogen signaling pathways, and nuclear factor-kappa B(NF-κB) signaling pathways. Additionally, the proteomics analysis highlighted important pathways such as peroxisome proliferator-activated receptor(PPAR) signaling pathways, mitogen-activated protein kinase(MAPK) signaling pathways, and β-alanine metabolism. The combined approaches of network pharmacology and proteomics have revealed that the mechanism by which YRJ improves osteoporosis may be closely related to the regulation of inflammation, osteoblast, and osteoclast metabolic pathways. The main pathways involved include the NF-κB signaling pathways, MAPK signaling pathways, and PPAR signaling pathways, among others.

This study aims to investigate the mechanism of Xueshuantong Injection(XST) on pulmonary fibrosis induced by bleomycin(BLM) in rats based on the coagulation cascade pathway. Sixty SD rats were randomly divided into sham surgery group,model group, pirfenidone(PFD, 50 mg·kg~(-1)) group, and 27, 54, and 81 mg·kg~(-1) XST groups. The rat model of pulmonary fibrosis was established by intratracheal injection of BLM(5 mg·kg~(-1)). After 24 hours, the administration groups were given corresponding drugs, while the sham surgery group and model group were given equal volumes of saline. On the 28th day, samples were collected,and the imaging and collagen fiber changes in the lungs of rats were observed. Immunofluorescence(IF) method was used to detect the expression level of alpha-smooth muscle actin(α-SMA), collagen Ⅰ(Col-Ⅰ), E-cadherin(E-cad), and vimentin(Vim). Western blot was used to determine the protein expression of α-SMA, Col-Ⅰ, Vim, and E-cad. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of prothrombin fragment(F1 + 2), thrombin-antithrombin complex(TAT), soluble fibrin monomer complex(SFMC), and rat fibrinogen degradation products(FDP) in rat lung tissue. Finally, the mRNA and protein levels of protease activated receptor 1(PAR-1) were detected by RT-qPCR, western blot, and IF. Compared with the model group, the scanning of the lungs of rats receiving XST treatment also exhibited patchy and non-homogeneous shadows, but these shadows were less dense than those in the model group. At the same time, there was a significant decrease in Col-Ⅰ fibers in the lungs of rats, and XST could inhibit epithelial-mesenchymal transition(EMT) and downregulate α-SMA and Col-Ⅰ protein expression. In the aspect of the coagulation system, administration of 81 mg·kg~(-1) XST significantly reduced the levels of SFMC and FDP. Meanwhile, 81 mg·kg~(-1) XST significantly downregulated the mRNA and protein levels of PAR-1. XST has an anti-pulmonary fibrosis effect in rats, and its mechanism may be related to the downregulation of PAR-1 to rebalance the coagulation cascade pathway.

A liquid chromatography-tandem mass spectrometry method was established and validated for determining the concentrations of costunolide(CO), piperine(PI), agarotetrol(AG), glycyrrhizic acid(GL), vanillic acid(VA), and glycyrrhetinic acid(GA) in rat plasma. This method was then applied to the toxicokinetic study of these six compounds in rats with chronic cerebral ischemia(CCI) following multiple oral doses of Zhachong Shisanwei Pills. Finally, the effects of continuous multiple-dose administration of Zhachong Shisanwei Pills on the liver of CCI rats were investigated. The results showed that after oral administration of different doses of Zhachong Shisanwei Pills, the in vivo exposure of AG, VA, and GA was relatively high, with AUC_(0-∞) values ranging from 604.0-2 494.2, 1 305.4-4 634.5, and 2 177.5-4 045.7 h·ng·mL~(-1), respectively, while the exposure of CO, PI, and GL was relatively low, with AUC_(0-∞) values ranging from 37.8-238.2, 2.4-17.0, and 146.9-408.5 h·ng·mL~(-1), respectively. The C_(max) and AUC_(0-∞) of the six compounds were positively correlated with the administered dose. The T_(max) of PI and AG ranged from 0.3 to 2.0 h, their T_(1/2) ranged from 0.8 to 2.9 h, and their mean residence time(MRT) ranged from 1.0 to 3.7 h. The T_(max) of GL and VA was shorter(0.4-1.9 h), while their T_(1/2)(2.6-5.9 h) and MRT(2.5-8.5 h) were longer. Both CO and GA exhibited a bimodal phenomenon, with T_(max) ranging from 1.6 to 6.6 h, T_(1/2) ranging from 2.8 to 7.7 h, and MRT ranging from 4.1 to 12.9 h. Liver histopathology after 28 days of continuous multiple-dose administration of Zhachong Shisanwei Pills showed that the liver tissue remained normal at a low dose(crude drug 0.8 g·kg~(-1), approximately 5 times the clinical equivalent dose). However, as the dose increased(crude drug 1.1-3.0 g·kg~(-1), 6.9-18.8 times the clinical equivalent dose), varying degrees of liver damage were observed. Blood biochemical tests revealed no significant changes in the serum levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), alkaline phosphatase(ALP), and total bile acid(TBA) in CCI rats from administration groups 1 to 3(crude drug 0.8, 1.1, 1.5 g·kg~(-1)). However, ALT, AST, ALP, and TBA levels in groups 4 and 5(crude drug 2.1, 3.0 g·kg~(-1)) showed significant increases. This study preliminarily elucidated the toxicokinetic characteristics of the six compounds in Zhachong Shisanwei Pills and their effects on liver tissue in CCI rats, providing data as a reference for clinical use.

Ethnopharmacological relevance: Pulmonary fibrosis (PF) is a chronic, progressive, and often fatal interstitial lung disease. Traditional Chinese medicine formulations and their active ingredients have shown potential in the treatment of PF. Panax notoginseng saponin (PNS) is extracted from the widely used traditional Chinese medicinal herb Panax notoginseng (Burkill) F. H. Chen, exhibiting therapeutic effects in pulmonary diseases treatment. Aim of the study: This study aimed to investigate the effects and elucidate possible potential mechanisms of PNS on bleomycin (BLM)-induced PF in rats. Materials and methods: PF was induced in rats by intratracheal administration of bleomycin (BLM, 5 mg/kg). After disease model induction, the rats were treated with PNS (50, 100, or 200 mg/kg per day) or pirfenidone (PFD, 50 mg/kg per day) for 28 days. Lung function, histopathological changes, collagen deposition, and E- and Ncadherin levels in lung tissue were evaluated. The mechanism of action of PNS was investigated using tandem mass tag-based quantitative proteomics analysis. Immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), and Western blot analysis were performed to verify the proteomic results. Results: PNS treatment improved lung function, ameliorated the BLM-induced increase in the lung coefficient, attenuated the degree of alveolar inflammation and fibrosis, and reduced the elevated collagen level in PF rats. PNS treatment also down-regulated the expression of N-cadherin while up-regulating the expression of E-cadherin. Proteomic and bioinformatic analyses revealed that the renin-angiotensin system (RAS) was closely related to the therapeutic effect of PNS. Immunohistochemistry, Western blot, and ELISA results indicated that PNS exerted its anti-fibrotic effect via regulation of the balance between the angiotensin-converting enzyme (ACE)angiotensin (Ang)II-AngII receptor type 1 (AT1R) and ACE2-Ang(1-7)-MasR axes. Conclusions: PNS ameliorates BLM-induced PF in rats by modulating the RAS homeostasis, and is a new potential therapeutic agent for PF.

Background: Luteolin-7-O-glucuronide (L7Gn) is a flavonoid isolated from numerous traditional Chinese herbal medicines that exerts anti-inflammatory effects. Previous research has revealed that aerosol inhalation is the most straightforward way of administration for the delivery of respiratory agents. Thus far, the impact of aerosol inhalation of L7Gn on lung inflammation and the underlying mechanisms remain unknown. Methods: The real-time particle size for L7Gn aerosol inhalation was detected by the Spraytec spray droplet size measurement system, including transmission and size diameters. The acute lung injury (ALI) rat model was induced by aerosol inhalation of LPS to evaluate the protective effect of L7Gn. The inhibitory effect of NLRP3 inflammasome activation assays was conducted in LPS-induced MH-S cells. Elisa, Western blotting, and RT-PCR were utilized to investigate the expression of NLRP3 inflammasome-relevant proteins and genes. Results: In this study, we found that inhalation of L7Gn aerosol significantly reduced pulmonary injury by inhibiting inflammatory infiltration and enhancing lung function. Meanwhile, the NLR family pyrin domain containing 3 (NLRP3) inflammasome was activated dramatically, accompanied by upregulated expression of IL-1β and IL-18, both in the ALI rat model and in LPS-induced MH-S cells. Moreover, L7Gn was found to significantly downregulate the expression of NLRP3, ASC, caspase-1, and cleaved caspase-1, which are critical components of the NLRP3 inflammasome, as well as the expression of IL-1β and IL-18. Conclusions: Based on our findings, L7Gn could exert anti-inflammatory effects by inhibiting NLRP3 inflammasome activation, which may emerge as potential therapeutic agents for the treatment of ALI.

目的 基于醒脑静雾化吸入给药粒径分布,多次雾化吸入给药黏膜过敏及刺激性,观察醒脑静雾化吸入给药的解热作用,为醒脑静临床雾化吸入给药提供依据和基础.方法 采用激光衍射法检测醒脑静雾化产生的粒径;通过连续多次雾化吸入给药观察醒脑静雾化吸入给药对大鼠黏膜刺激性;通过豚鼠全身主动过敏实验考察醒脑静雾化吸入给药过敏反应;采用干酵母致热模型观察醒脑静雾化吸入溶液解热作用.结果 粒径分布结果显示,醒脑静雾化最小粒径为0.117μm,90％粒径在10 μm以下左右,Dv(50)=4.76 μm提示醒脑静雾化后能够进入各级支气管及肺.刺激实验结果显示醒脑静雾化给药对大鼠肺灌洗液白细胞及其分类无显著影响,病理结果显示对鼻黏膜、咽喉、气管和肺部无明显影响.过敏结果显示醒脑静雾化吸入给药豚鼠全身主动过敏反应阴性.解热实验结果表明,醒脑静吸入给药能够明显抑制干酵母诱导发热模型大鼠的体温升高,能够降低下丘脑前列腺素E2(prostaglandin E2,PGE2)及环磷酸腺苷(cyclic adeno-sine monophosphate,cAMP)的含量和血清中肿瘤坏死因子(tumor necrosis factor-α,TNF-α)的含量.结论 醒脑静雾化粒径能够进入各级支气管及肺,雾化吸入后对大鼠粘膜无刺激性,豚鼠全身主动过敏反应阴性.对干酵母诱导的大鼠发热模型具有解热作用,其作用机理可能与其降低下丘脑PGE2及cAMP和血清中TNF-α的含量有关.

Objective: To explore the characteristics and mechanism of Zhentongji Capsule on hyperlipidemia model rats by high-resolution untarget metabonomics. Methods: Forty SPF SD rats were used in this experiment. Six animals were randomly selected as the control group and given normal diet， and all the other animals were given high-fat diet. After 4 weeks with successful modeling， they were divided into model group and administration group. The administration group was divided into high， medium and low dose groups. The Administration group was given Zhentongji Capsule once a day for 8 weeks via i.g， with the dosage of 0.432， 0.216， 0.108 g·kg -1 . After the last administration， the animals were anesthetized， blood was collected from abdominal aorta， the serum was centrifuged， and the blood lipid level in the serum was detected by automatic biochemical instrument. Some liver tissues were taken for the study of high-resolution untarget metabonomics. Results: After administration of Zhentongji Capsule， the levels of triglyceride， cholesterol， low-density lipoprotein and high-density lipoprotein in high and medium doses were significantly lower than those in the model group， and the difference was statistically significant. Compared with the control group， 138 different metabolites were detected in the model group. Compared with the model group， 123 different metabolites were detected in the administration group， 24 of 72 different metabolites in positive ion mode were corrected after administration， and 9 of 51 different metabolites in negative ion mode were corrected after administration. The metabolic process involves multiple signal pathways， mainly including mammalian target of rapamycin （mTOR）signal pathway， cyclic guanosine 3’，5’-monophosphate-dependent protein kinase （cGMP-PKG） signal pathway， ForkheadBoxProteinO （FoxO） signal pathway， protein digestion and absorption， pyrimidine metabolism， microbial digestion and absorption， glycerol phospholipid metabolism， amino acid biosynthesis， purine metabolism， etc. Conclusion: Zhentongji Capsule can effectively decrease the blood lipid level of hyperlipidemia rats， which was closely related to its ability to regulate purine metabolism， pyrimidine metabolism， glycerophosphatide metabolism， mTOR signal pathway and cGMP-PKG signal pathway.

Purpose: This study aimed to investigate the main pharmacological action and underlying mechanisms of Jin Gu Lian Capsule (JGL) against rheumatoid arthritis (RA) based on network pharmacology and experimental verification.Methods: Network pharmacology approaches were performed to explore the core active compounds of JGL, key therapeutic targets, and signaling pathways. Molecular docking was used to predict the binding affinity of compounds with targets. In vivo experiments were undertaken to validate the findings from network analysis.Results: A total of 52 targets were identified as candidate JGL targets for RA. Sixteen ingredients were identified as the core active compounds, including, quercetin, myricetin, salidroside, etc. Interleukin-1 beta (IL1B), transcription factor AP-1 (JUN), growth-regulated alpha protein (CXCL1), C-X-C motif chemokine (CXCL)3, CXCL2, signal transducer and activator of transcription 1 (STAT1), prostaglandin G/H synthase 2 (PTGS2), matrix metalloproteinase (MMP)1, inhibitor of nuclear factor kappa-B kinase subunit beta (IKBKB) and transcription factor p65 (RELA) were obtained as the key therapeutic targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis showed that the efficacy of JGL was functionally involved in regulating immune-mediated inflammation, in which IL-17/NF-kappa B signaling was recommended as one of the main pathways. Molecular docking suggested that the core active compounds bound strongly to their respective targets. Experimentally, JGL treatment mitigated inflammation, showed analgesic activity, and ameliorated collagen-induced arthritis. Enzyme-linked immunosorbent assay showed that JGL effectively reduced the serum levels of cytokines, chemokines, and MMPs. Immunohistochemistry staining showed that JGL markedly reduced the expression of the targets in IL-17/NF-kappa B pathway including IL-17A, IL-17RA, NF-kappa B p65, C-X-C motif ligand 2, MMP1 and MMP13. Conclusion: This investigation provided evidence that JGL may alleviate RA symptoms by partially inhibiting the immune-mediated inflammation via IL-17/NF-kappa B pathway.

目的:制备咖啡酸分子烙印聚合物(MIP)并特异性地去除金银花提取物中的咖啡酸,考察咖啡酸去除前后金银花提取物抑制前列腺素E2(PGE2)释放的变化,从整体上评价其抗炎活性.方法:采用溶胶-凝胶法,以粒径为 62～105 μm的二氧化硅微珠为载体、咖啡酸为模板分子、(3-氨丙基)三乙氧基硅烷为功能单体、四乙氧基硅烷为交联剂、四氢呋喃为溶剂,合成了咖啡酸MIP.以此MIP作为液相色谱固定相,以甲醇-乙酸(500∶1和 9∶1)为流动相,特异性去除了金银花提取物中的咖啡酸.通过脂多糖刺激巨噬细胞RAW264.7 释放PGE2 实验评价去除咖啡酸前后金银花提取物的抗炎作用.结果:溶胶-凝胶法制备的咖啡酸MIP能够从复杂体系中特异性地分离和富集微量的咖啡酸,对咖啡酸的容量因子和烙印效率分别为 15.8 和 9.7,最终从金银花提取物 2.6 g中分离了咖啡酸146 μg,纯度为 92%,回收率为 89%.金银花提取物和去除咖啡酸的提取物在质量浓度为 100、200、400 μg·mL-1时对脂多糖诱导的RAW264.7 细胞释放PGE2 的抑制率分别为 5.8%、35.6%、62.5%和 5.4%、13.3%、57.5%,去除微量咖啡酸以后的提取物在 3 个质量浓度时的抑制率较金银花提取物均有所降低.结论:咖啡酸是金银花提取物抗炎活性的一个重要成分,咖啡酸分子烙印聚合物可以实现金银花提取物中微量咖啡酸的特异性分离,分子烙印技术能够在保证中药完整性的前提下评价中药的药理活性.

目的 基于细胞色素P450(CYP450)系统研究乌头碱配伍鞣花酸、甘草苷减毒机制.方法 将HepG2细胞分为空白组、乌头碱组、鞣花酸组、甘草苷组、乌头碱+鞣花酸组、乌头碱+甘草苷组、乌头碱+鞣花酸+甘草苷组,CCK8法和LDH法分别检测细胞活力和细胞毒性,高内涵分析技术检测细胞数目、DNA和活性氧(ROS)含量及线粒体膜电位(MMP),RT-PCR和Western blot检测CYP1A2、CYP2C9和CYP3A4 mRNA及蛋白表达.结果 与空白组比较,乌头碱组细胞ROS含量显著增加,MMP显著降低(P<0.05),乌头碱+鞣花酸+甘草苷组细胞ROS、MMP差异无统计学意义(P>0.05).RT-PCR和Western blot结果显示,与空白组比较,乌头碱组细胞CYP1A2、CYP3A4 mRNA和蛋白表达显著降低(P<0.05),甘草苷组细胞CYP1A2、CYP2C9、CYP3A4 mRNA和蛋白表达显著升高(P<0.01,P<0.05);与乌头碱组比较,乌头碱+鞣花酸+甘草苷组细胞CYP1A2、CYP3A4 mRNA和蛋白表达显著升高(P<0.01,P<0.05,P<0.001).结论 乌头碱配伍鞣花酸、甘草苷可上调CYP1A2、CYP3A4表达,减少乌头碱在体内蓄积时间,起到减毒作用.

[目的]研究香砂平胃颗粒对正常小鼠及胃肠动力障碍模型小鼠胃排空、肠推进和血清中胃动素(MOT)、胃泌素(GAS)、生长抑素(SS)、P物质(SP)和生长激素释放肽(Ghrelin)含量的影响.[方法]灌胃给予正常小鼠及阿托品所致胃肠动力障碍模型小鼠香砂平胃颗粒高、中、低剂量(2.6 g/kg、1.3 g/kg、0.65 g/kg),采用称重法计算小鼠胃残留率和小肠炭末推进法计算小鼠肠推进率,进一步应用ELISA方法检测小鼠血清中MOT、GAS、SS、SP及Ghrelin的含量,并利用RT-PCR技术检测下丘脑中Ghrelin mRNA与生长激素促分泌素受体(GHSR)mRNA转录水平.[结果]在胃肠运动方面,香砂平胃颗粒高剂量组能显著促进正常小鼠的胃排空(P<0.05),并能拮抗阿托品所致小鼠的胃排空抑制作用(P<0.05),香砂平胃颗粒高、中、低剂量组均能显著拮抗胃肠动力障碍模型小鼠的小肠推进抑制作用(P<0.05或P<0.01);在胃肠激素方面,香砂平胃颗粒高剂量组能提高正常小鼠及胃肠动力障碍模型小鼠血清中MOT和GAS水平(P<0.05或P<0.01),且能上调胃肠动力障碍模型小鼠血清中Ghrelin含量与下丘脑中Ghrelin mRNA转录水平(P<0.01).[结论]香砂平胃颗粒能提高正常小鼠与胃肠动力障碍模型小鼠的胃肠动力,其作用机制可能与增加血清中MOT、GAS与Ghrelin含量有关.

Objective:To investigate the effects of Guiling Gao on body temperature, gastrointestinal motility, gastrointestinal hormones, Th1/Th2 cytokines and water metabolism in rats with damp-heat syndrome.Methods:Totally 60 SD rats were randomly divided into control group, model group, mosapride group, Guiling Gao low dose group (3.4 g/kg), medium dose group (6.8 g/kg) and high dose group (13.6 g/kg) according to random number table method, with 10 rats in each group. Except for the blank group, the other groups adopted the method of "environmental factors + fat and sweet diet + biological factors" to prepare the rat model of damp heat syndrome of febrile diseases. After modeling, they were administered by gavage for 7 days. During the experiment, the general state, body weight and body temperature were observed, the gastric residue rate of rats was calculated by weighing method, the intestinal propulsion rate of rats was calculated by charcoal propulsion method, and the levels of serum motilin (MTL), gastrin (GAS), somatostatin (SS), substance P (SP),IL-4 and interferon-γ (IFN-γ) were detected by ELISA, and the changes of aquaporin 3 (AQP3) mRNA transcription level were detected by real-time PCR.Results:Compared with the model group, the weight of rats in Guiling Gao high dose group increased after experiment of 22 days ( P<0.05), and body temperature of rats in Guiling Gao medium and high dose group decreased in 19-20 day ( P<0.01); and the gastric emptying rate and the small intestine propulsion rate of small intestine in Guiling Gao medium and high dose group increased significantly ( P<0.01 or P<0.05); the serum MTL, GAS and SP levels increased ( P<0.01 or P<0.05), and SS decreased ( P<0.01 or P<0.05) in the Guiling Gao medium and high dose groups; The levels of IL-4, IFN-γ and IFN-γ/IL-4 ratio decreased ( P<0.01); The expression of AQP3 mRNA (1.16 ± 0.25 vs. 0.23 ± 0.01) in the Guiling Gao high dose group was up-regulated ( P<0.01). Conclusions:Guiling Gao can effectively improve the activity state of damp-heat syndrome model rats caused by complex factors. This mechanism may be related to enhancing gastrointestinal movement, increasing gastrointestinal hormone secretion, restoring the dynamic balance of immune system Th1/Th2 and promoting the transport of water from intestinal cavity.

Background:A. chinense frequently used in Miao medicine to treat rheumatic diseases. However, as a famous toxic herb, Alangium chinense and its representative components exhibit ineluctable neurotoxicity, thus creating significant challenges for clinical application. The combined application with compatible herbs in Jin-Gu-Lian formula attenuates such neurotoxicity according to the compatible principle of traditional Chinese medicines. Purpose: We aimed to investigate the detoxification of the compatible herbs in Jin-Gu-Lian formula on A. chinense-induced neurotoxicity and investigate its mechanism. Methods: Neurobehavioral and pathohistological analysis were used to determine the neurotoxicity in rats administered with A. chinense extract (AC), extract of compatible herbs in Jin-Gu-Lian formula (CH) and combination of AC with CH for 14 days. The mechanism underlying the reduction of toxicity by combination with CH was assessed by enzyme-linked immunosorbent assays, spectrophotometric assays, liquid chromatography tandem-mass spectrometry and real-time reverse transcription-quantitative polymerase chain reaction. Results: Compatible herbs attenuated the AC-induced neurotoxicity as evidenced by increased locomotor activity, enhanced grip strength, the decreased frequency of AC-induced morphological damage in neurons, as well as a reduction of neuron-specific enolase (NSE) and neurofilament light chain (NEFL) levels. The combination of AC and CH ameliorated AC-induced oxidative damage by modulating the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC). AC treatment significantly reduced the levels of monoamine and acetylcholine neurotransmitters in the brains of rats, including acetylcholine (Ach), dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), norepinephrine (NE), and serotonin (5-HT). Combined AC and CH treatment regulated the abnormal concentrations and metabolisms of neurotransmitters. Pharmacokinetic studies showed that the co-administration of AC and CH significantly decreased plasma exposure levels of two main components of AC, as evidenced by the reduction of maximum plasma concentration (Cmax), area under the plasma concentration-time curve (AUC) compared to AC. In addition, the AC-induced downregulation in mRNA expression of cytochrome P450 enzymes was significantly reduced in response to combined AC and CH treatment. Conclusion: Compatible herbs in Jin-Gu-Lian formula alleviated the neurotoxicity induced by A. chinense by ameliorating oxidative damage, preventing abnormality of neurotransmitters and modulating pharmacokinetics.

目的:基于代谢组学探究阿托伐他汀联合臻通集胶囊治疗高脂血症模型大鼠的疗效和作用机制.方法:SD大鼠随机取6只作为对照组,正常饮食、饮水.其余动物均给予高脂饲料,连续4周后,甘油三酯(TG)和胆固醇(CHO)明显升高视为模型成功;根据CHO水平进行分组,分为模型组、西药组(阿托伐他汀钙片,0.9 mg/kg)、中药组(臻通集胶囊,0.216 g/kg)、中西医联合组(二者联用).连续给药8周后,检测各组动物血清中的血脂水平,肝脏组织用于高分辨非靶代谢组学的研究.结果:模型组TG、CHO、高密度脂蛋白(HDL)和低密度脂蛋白(LDL)水平较对照组显著升高(P＜0.05);与模型组相比,西药组、中药组和中西医联合组血脂水平显著降低(P＜0.05):与对照组相比,模型组共检测出138个差异代谢物;与模型组相比,西药组共检测出64个差异代谢物,中药组共检测出123个差异代谢物,中西医联合组共检出106个差异代谢物;中药组与西药组相比,共检出101个差异代谢物;经相关性分析和KEGG通路分析,主要涉及衰老、脂质代谢、氨基酸代谢、膜转运等通路.结论:阿托伐他汀联合臻通集胶囊能够有效地改善高血脂大鼠的血脂水平,代谢组学分析示其可能通过调节cGMP-PKG信号通路相关因子的表达,达到抑制肝脏细胞坏死的目的.

Background Systemic sclerosis (SSc; also known as “scleroderma”) is an autoimmune disorder characterized by extensive fibrosis, vascular changes, and immunologic dysregulation. Baicalein (phenolic flavonoid derived from Scutellaria baicalensis Georgi) has been used to treat the pathological processes of various fibrotic and inflammatory diseases. In this study, we investigated the effect of baicalein on the major pathologic characteristics of SSc: fibrosis, B-cell abnormalities, and inflammation. Methods The effect of baicalein on collagen accumulation and expression of fibrogenic markers in human dermal fibroblasts were analyzed. SSc mice were produced by injecting bleomycin and treated with baicalein (25, 50, or 100 mg/kg). The antifibrotic features of baicalein and its mechanisms were investigated by histologic examination, hydroxyproline assay, enzyme-linked immunosorbent assay, western blotting and flow cytometry. Results Baicalein (5–120 μM) significantly inhibited the accumulation of the extracellular matrix and fibroblast activation in transforming growth factor (TGF)-β1- and platelet derived growth factor (PDGF)-induced human dermal fibroblasts, as evidenced by abrogated deposition of total collagen, decreased secretion of soluble collagen, reduced collagen contraction capability and downregulation of various fibrogenesis molecules. In a bleomycin-induced model of dermal fibrosis in mice, baicalein (25–100 mg/kg) restored dermal architecture, ameliorated inflammatory infiltrates, and attenuated dermal thickness and collagen accumulation in a dose-dependent manner. According to flow cytometry, baicalein reduced the proportion of B cells (B220 + lymphocytes) and increased the proportion of memory B cells (B220 + CD27 + lymphocytes) in the spleens of bleomycin-induced mice. Baicalein treatment potently attenuated serum levels of cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-17A, tumor necrosis factor-α), chemokines (monocyte chemoattractant protein-1, macrophage inflammatory protein-1 beta) and autoantibodies (anti-scleroderma 70 (Scl-70), anti-polymyositis-scleroderma (PM-Scl), anti-centromeres, anti-double stranded DNA (dsDNA). In addition, baicalein treatment can significantly inhibit the activation of TGF-β1 signaling in dermal fibroblasts and bleomycin-induce mice of SSc, evidenced by reducing the expression of TGF-β1 and IL-11, as well as inhibiting both small mother against decapentaplegic homolog 3 (SMAD3) and extracellular signal-related kinase (ERK) activation. Conclusions These findings suggest that baicalein has therapeutic potential against SSc, exerting modulating B-cell abnormalities, anti-inflammatory effects, and antifibrosis.

Objective To study the therapeutic effect of Tanreqing Injection(痰热清注射液)on chronic obstructive pulmonary disease(COPD)model rats,and explore its possible mechanism by tandem mass tags(TMT)quantitative protein omics.Methods Wistar rats were randomly divided into control group,model group,dexamethasone(4 mg/kg)group and Tanreqing Injection high-,medium-and low-dose(4,2,1 g/kg)groups.Except for the control group,COPD models were prepared in other groups,and the corresponding drugs were given to each group.After 90 d of continuous administration,lung function and cytokine level in alveolar lavage fluid were detected.Hematoxylin-eosin and picrosirius red staining were used to observe the pathological changes of lung tissue.The differential proteins in lung tissue of rats in control group,model group and Tanreqing Injection medium-dose group were analyzed by TMT quantitative protein omics.Results Compared with control group,the autonomic activity and mental state of rats in model group were decreased,the compliance of lung function was decreased(P＜0.001),the resistance was increased(P＜0.001),inflammatory cells infiltrated in lung tissue,alveolar cavity expanded and collagen fibers deposited in airway wall,which indicated that the COPD model was successfully prepared.Compared with model group,the lung function of rats in Tanreqing Injection group was significantly improved(P＜0.01,0.001),and the levels of inflammatory factors in alveolar lavage fluid were significantly decreased(P＜0.05,0.01,0.001).Proteomic analysis showed that there were 61 differential proteins in Tanreqing Injection medium-dose group and model group,which were closely related to cell necrosis,apoptosis,autophagy and other signal pathways.Conclusion Tanreqing Injection can obviously improve the lung function and other symptoms of COPD model rats,and its mechanism is closely related to signal pathways such as cell necrosis,apoptosis and autophagy.

Tibetan medicine is one of the oldest traditional medicine systems in the world. Taking the Ruyi Zhenbao tablet (RYZB) as an example, which is a widely used classic oral Tibetan medicine, this article discusses the pharmacokinetics of single administration and long-term treatment and analyzed its metabolic properties and tissue distribution in vivo. After single administration, blood samples were collected before administration and at different time points after administration in different groups of rats. In the study of long-term treatment effects, blood samples were collected from the animals in each group on days 1, 15, and 30 and on day 15 after withdrawal. The results showed that after a single administration, the dose change had no significant effect on the T1/2 and Tmax of agarotetrol, isoliquiritigenin, and piperine (p > 0.05). There was a certain correlation between the increase in AUC0-t and the Cmax of agarotetrol, isoliquiritigenin, piperine, and the increase in dosage, with a dose range of 0.225–0.900 g/kg. There were no significant differences in Cmax and AUC0-t of ferulic acid at different doses (p > 0.05). Meanwhile, there was no significant sex-based difference in the pharmacokinetic parameters of these four components in rats. After long-term administration, the distribution agarotetrol in various tissues of rats was kidney > liver > heart > brain; the tissue distribution in low- and medium-dose groups of isoliquiritigenin was liver > kidney > heart > brain, and in the high-dose group, kidney > liver > heart > brain. The tissue distribution of piperine in each dose group was liver > kidney > heart > brain, and that of ferulic acid in each dose group was kidney > liver > heart > brain. Through the establishment of the previously developed methodology, the pharmacokinetic properties of RYZB were analyzed after a single administration and long-term administration. Our findings confirmed this approach for the exploration and establishment of a pharmacokinetic evaluation of Tibetan medicine, to support its guiding role in clinical application, but also to accelerate research into Tibetan medicine theory and medicine and to provide a solid foundation for the translation of Tibetan medicine throughout the world.

目的:探讨复方苦参注射液与血必净注射液对大鼠急性肺损伤的作用.方法:将Wistar大鼠随机分为正常组、模型组、阳性对照(地塞米松注射液)组、血必净注射液组及苦参高、中、低剂量组.除正常组外,其余各组均采用脂多糖(LPS)建立急性肺损伤大鼠模型,连续3d.造模后立即给药,末次造模后15 h给予第4次给药.给药后1h处死动物,取肺灌洗液和肺组织,采用酶联免疫吸附试验(ELISA)法检测肺灌洗液和组织匀浆中白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子(TNF-α)和核因子(NF)-κB的含量,苏木精—伊红(HE)染色观察肺组织病理学变化.结果:与正常组比较,模型组肺组织炎性细胞浸润程度升高,阳性对照组、血必净注射液组、苦参高剂量组肺组织炎性细胞浸润程度低于模型组.与模型组比较,苦参高、中剂量组和血必净注射液组肺灌洗液和肺组织中IL-1β、IL-6、NF-κB和TNF-α水平降低(均P＜0.05);与血必净注射组比较,苦参注射液中剂量组肺灌洗液和肺组织中NF-κB、TNF-α水平降低(P＜0.05).结论:复方苦参注射液和血必净注射液在治疗急性肺损伤方面均有一定的效果,能够有效抑制炎症反应,且前者的效果更为明显.