Infection with influenza A virus (IAV) may trigger excessive inflammatory responses, leading to severe viral pneumonia and accelerating disease progression. Therefore, controlling these excessive inflammatory responses is crucial for the prevention and treatment of pneumonia caused by IAV. Berberine (BBR), an isoquinoline alkaloid extracted from traditional Chinese medicine, possesses extensive pharmacological activities. However, its immunoregulatory effects and molecular mechanisms in the context of IAV infection require further investigation. This study explored the impact of BBR on macrophage pyroptosis and inflammatory responses induced by IAV infection. Our findings revealed that BBR effectively inhibits the release of IL-1β and TNF-α induced by IAV infection and suppresses gasdermin D (GSDMD)-mediated pyroptosis in a dose-dependent manner. Further research indicates that BBR alleviates macrophage pyroptosis and inflammatory responses in IAV-infected cells by reducing the release of mitochondrial reactive oxygen species (mtROS), inhibiting mitochondrial antiviral signaling protein (MAVS) expression and blocking the activation of the NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome. Experiments using siRNA to knockdown MAVS further confirmed the pivotal role of MAVS in BBR’s inhibition of IAV-induced macrophage pyroptosis. This study provides a scientific basis for the application of BBR as an anti-inflammatory drug in the treatment of inflammatory diseases caused by IAV infection and directs future research endeavors.

The rapeseed meal, a type of residual by-product of rapeseed oil production was used as the precursor to prepare nitrogen self-doping carbon dots RM-CDs through an easy hydrothermal process. Thanks to the introduction of nitrogen element and oxygen-containing functional groups, RM-CDs had a fluorescence quantum yield of 18.6 %. The RM-CDs demonstrated highly stable to the ionic strength, pH, and ultraviolet radiation. However, their bright blue fluorescence was significantly weakened by baicalein owing to the inner-filter effect and static quenching effect. As a result, RM-CDs could be used as the fluorescent sensing materials for baicalein detection with a wide range of linear response (0.5-170 mu M) as well as low limit of detection (0.046 mu M). Moreover, the as- developed sensing platform was also applicable to detect baicalein in actual samples. The developed RM-CDs was sustainably prepared from the biomass by-product of rapeseed meal, this approach expanded the application scopes of CDs.

In this study, the efficacy of Fatigue, Resistance, Ambulation, Illness and Loss of weight (FRAIL) management combined with acetylcysteine (AChE) in blood purification therapy for sepsis (SS) patients was investigated. Seventy-eight SS patients undergoing blood purification were randomized to an observation group (FRAIL + AChE, n=42) or a control group (conventional care, n=36). FRAIL score, APACHE II score, vital signs (heart rate, mean arterial pressure, oxygen saturation), inflammatory factors oxidative stress indexes and nutritional proteins were assessed before and after the intervention in both groups. The results of the study showed that after the intervention, the FRAIL score and APACHE II score were reduced more significantly in the observation group (P<0.05). In addition, vital signs improved more significantly in the observation group. After the intervention, all inflammatory factors in the observation group were lower than those in the control group, while nutritional proteins were higher than those in the control group (P<0.05). In addition, oxidative stress injury was also more favorable in the observation group than in the control group after the intervention. These results suggest that FRAIL management combined with AChE may play a significant role in SS treatment.

Objective To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD). Methods Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice. Results The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group ( P <0.05). Serum DLA was significantly lower in the YD group than HCD-P group ( P <0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group ( P <0.01). There was no statistical significance in the serum level of DAO among groups ( P >0.05). Conclusions YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.

Renal fibrosis (RF) is the common pathway for a variety of chronic kidney diseases that progress to end-stage renal disease. Chitosan oligosaccharide (COS) has been identified as possessing many health functions. However, it is not clear whether COS can prevent RF. The purpose of this paper was to explore the action and mechanism of COS in alleviating RF. First, an acute unilateral ureteral obstruction operation (UUO) in male BALB/c mice was performed to induce RF, and COS or fosinopril (positive control drug) were administered for 7 consecutive days. Data from our experiments indicated that COS treatment can significantly alleviate kidney injury and decrease the levels of blood urea nitrogen (BUN) and serum creatinine (SCr) in the UUO mouse model. More importantly, our results show that COS can reduce collagen deposition and decrease the expression of fibrosis proteins, such as collagen IV, fibronectin, collagen I, α-smooth muscle actin (α-SMA) and E-cadherin, ameliorating experimental renal fibrosis in vivo. In addition, we also found that COS suppressed oxidative stress and inflammation in RF model mice. Further studies indicated that the mechanism by which COS alleviates renal fibrosis is closely related to the regulation of the TGF-β1/Smad pathway. COS has a therapeutic effect on ameliorating renal fibrosis similar to that of the positive control drug fosinopril. Taken together, COS can alleviate renal fibrosis induced by UUO by reducing oxidative stress damage and regulating the TGF-β1/Smad pathway.

目的 探讨苦参碱(MT)逆转肺癌A549顺铂耐药株活性中的作用及分子机制.方法 对数生长期人肺腺癌耐顺铂细胞株A549/DDP细胞,用不同浓度MT(1,4,16,32μmol·L-1)处理,采用噻唑蓝(MTT)法在24 h检测细胞增殖抑制率.将细胞分为空白对照组及MT组(16μmol·L-1),光学显微镜、Transwell及免疫荧光检测两组细胞的细胞形态、侵袭能力、E-钙粘蛋白(E-cadherin)及波形蛋白(vimentin)细胞免疫荧光变化.另将细胞分为空白对照组、MT组(16μmol·L-1MT)、顺铂(DDP)组(3×10-3μmol·L-1DDP)和MT+DDP组(16μmol·L-1MT+3×10-3μmol·L-1DDP),Western blotting法检测上皮间充质转化(EMT)表型相关标志物E-caherin、vimentin、slug、p-p65等蛋白表达,流式细胞仪检测细胞凋亡率.结果 MT能明显抑制A549/DDP细胞的增殖及侵袭能力,MT处理后细胞呈现上皮细胞样形态及上皮表型.此外,与对照组及DDP组比较,MT+DDP组细胞凋亡率明显升高(P<0.05),E-cadherin蛋白表达水平明显升高(P<0.05),vimentin、slug及p-p65蛋白表达水平明显降低(P<0.05).结论 苦参碱具有逆转肺癌顺铂耐药的作用,其机制与抑制肺癌细胞上皮间质转化及抑制核因子NF-κB信号通路有关.

目的 比较截断疗法经典方犀角地黄汤合银翘散及其单方对小鼠流感病毒性肺炎的治疗作用,探究犀角地黄汤和银翘散在截断疗法中的不同作用.方法 ICR小鼠和BALB/c小鼠,随机分为正常组、模型组、犀角地黄汤合银翘散组(简称合方组)、犀角地黄汤组、银翘散组、达菲组,除正常组外其余小鼠以流感病毒滴鼻感染,1 h后给药.各组分别以对应的水煎剂或溶液灌胃,每日1次,连续6 d.ICR小鼠观察14 d,记录小鼠体重及生存时间;BALB/c小鼠分别在感染后第2、4、6天取材,光镜下观察肺组织病理切片、RT-PCR法检测肺组织中甲型流感病毒核蛋白(IAV-NP)mR-NA水平、ELISA法检测肺组织肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平.结果 合方组可提高感染小鼠生存率,延长生存时间,推后死亡极期,效果优于犀角地黄汤组和银翘散组,而犀角地黄汤组优于银翘散组;合方组在第6天可减轻感染小鼠肺组织炎性病理改变,效果优于犀角地黄汤组和银翘散组,犀角地黄汤组比银翘散组病变轻.在第2、4、6天,银翘散组IAV-NP mRNA表达降低,与模型组比较差异有统计学意义(P<0.01),且在第4、6天,银翘散组IAV-NP mRNA表达低于合方组(P<0.01);第4、6天,犀角地黄汤组IAV-NP mRNA表达与模型组比较,差异无统计学意义.在第4、6天,合方组、犀角地黄汤组和银翘散组肺组织TNF-α和IL-6水平低于模型组(P<0.05、P<0.01),合方组肺组织TNF-α 和IL-6水平低于犀角地黄汤组和银翘散组(P<0.05、P<0.01).犀角地黄汤组第6天TNF-α水平低于银翘散组(P<0.05),而其第2、4天IL-6水平低于银翘散组(P<0.01).结论 银翘散以抗病毒作用为主,而犀角地黄汤以抗炎作用为主,两方合用可在病毒感染早期截断病邪深入,又在疾病极期扭转严重炎症,以达到治疗效果.

The present study investigated the sensitization of 5-fluorouracil (5-FU)-resistant colon cancer cells in vitro, using oxymatrine, a Chinese herb, and a quinolizidine alkaloid compound extracted from the root of Sophora flavescens. The HCT-8 colon cancer cell line and its 5-FU-resistant subline HCT-8/5-FU were treated with 5-FU and oxymatrine, alone or in combination, at various doses. The cells were subsequently assessed for changes in cell viability, apoptosis and morphology and analyzed by fluorescence microscopy and western blotting. The data demonstrated that HCT-8/5-FU markedly increased the dose of 5-FU required for the suppression of tumor cell viability (78.77±1.90 µg/ml vs. 9.20±0.96 µg/ml in parental HCT-8 cells), whereas HCT-8/5-FU induced the tumor cell epithelial-mesenchymal transition (EMT). By contrast, oxymatrine alone and in combination with 5-FU altered HCT-8/5-FU cell morphology, apoptosis and EMT phenotypes. The combination of oxymatrine and 5-FU reduced the protein expression of snail family transcriptional repressor 2 and vimentin, phosphorylated p65 and induced the expression of E-cadherin, by inhibiting the nuclear factor κB (NF-κB) signaling pathway. In conclusion, the data from the present study demonstrated that EMT was associated with 5-FU chemoresistance in HCT-8/5-FU colon cancer cells, and that oxymatrine treatment was able to reverse such resistance. Oxymatrine may regulate tumor cell EMT and inactivate the NF-κB signaling pathway, and may therefore serve as a potential therapeutic drug to reverse 5-FU resistance in colon cancer cells.

BACKGROUND Massive hepatocyte death is the core event in acute liver failure (ALF). Gasdermin D (GSDMD)-mediated pyroptosis is a type of highly inflammatory cell death. However, the role of hepatocyte pyroptosis and its mechanisms of expanding inflammatory responses in ALF are unclear. AIM To investigate the role and mechanisms of GSDMD-mediated hepatocyte pyroptosis through in vitro and in vivo experiments. METHODS The expression of pyroptosis pathway-associated proteins in liver tissues from ALF patients and a hepatocyte injury model was examined by Western blot. GSDMD short hairpin RNA (shRNA) was used to investigate the effects of downregulation of GSDMD on monocyte chemotactic protein 1 (MCP1) and its receptor CC chemokine receptor-2 (CCR2) in vitro. For in vivo experiments, we used GSDMD knockout mice to investigate the role and mechanism of GSDMD in a D-galactose/lipopolysaccharide (D-Galn/LPS)-induced ALF mouse model. RESULTS The levels of pyroptosis pathway-associated proteins in liver tissue from ALF patients and a hepatocyte injury model increased significantly. The level of GSDMD-N protein increased most obviously (P < 0.001). In vitro, downregulation of GSDMD by shRNA decreased the cell inhibition rate and the levels of MCP1/CCR2 proteins (P < 0.01). In vivo, GSDMD knockout dramatically eliminated inflammatory damage in the liver and improved the survival of D-Galn/LPS-induced ALF mice (P < 0.001). Unlike the mechanism of immune cell pyroptosis that involves releasing interleukin (IL)-1β and IL-18, GSDMD-mediated hepatocyte pyroptosis recruited macrophages via MCP1/CCR2 to aggravate hepatocyte death. However, this pathological process was inhibited after knocking down GSDMD. CONCLUSION GSDMD-mediated hepatocyte pyroptosis plays an important role in the pathogenesis of ALF, recruiting macrophages to release inflammatory mediators by upregulating MCP1/CCR2 and leading to expansion of the inflammatory responses. GSDMD knockout can reduce hepatocyte death and inflammatory responses, thus alleviating ALF.

Aging that refers the accumulation of genetic and physiology changes in cells and tissues over a lifetime has been shown a high risk of developing various complex diseases, such as neurodegenerative disease, cardiovascular disease and cancer. Over the past several decades, natural products have been demonstrated as anti-aging interveners via extending lifespan and preventing aging-associated disorders. In this study, we developed an integrated systems pharmacology infrastructure to uncover new indications for aging-associated disorders by natural products. Specifically, we incorporated 411 high-quality aging-associated human genes or human-orthologous genes from mus musculus (MM), saccharomyces cerevisiae (SC), caenorhabditis elegans (CE), and drosophila melanogaster (DM). We constructed a global drug-target network of natural products by integrating both experimental and computationally predicted drug-target interactions (DTI). We further built the statistical network models for identification of new anti-aging indications of natural products through integration of the curated aging-associated genes and drug-target network of natural products. High accuracy was achieved on the network models. We showcased several network-predicted anti-aging indications of four typical natural products (caffeic acid, metformin, myricetin, and resveratrol) with new mechanism-of-actions. In summary, this study offers a powerful systems pharmacology infrastructure to identify natural products for treatment of aging-associated disorders.

Severe influenza infections are featured by acute lung injury, a syndrome of pulmonary microvascular leak. A growing number of evidences have shown that the pulmonary microvascular endothelial cells (PMVEC) are critical target of influenza virus, promoting microvascular leak. It is reported that there are multiple mechanisms by which influenza virus could elicit increased pulmonary endothelial permeability, in both direct and indirect manners. Ezrin/radixin/moesin family proteins, the linkers between plasma membrane and actin cytoskeleton, have been reported to be involved in cell adhesion, motility and may modulate endothelial permeability. Studies have also shown that ERM is phosphorylated in response to various stimuli via p38MAPK, Rho/ROCK or PKC pathways. However, it is unclear that whether influenza infection could induce ERM phosphorylation and its relocalization. In the present study, we have found that there are cytoskeletal reorganization and permeability increases in the course of influenza virus infection, accompanied by upregulated levels of p-ERM. p-ERM's aggregation along the periphery of PMVEC upon influenza virus infection was detected via confocal microscopy. Furthermore, we sought to determine the role of p38MAPK, Rho/ROCK and PKC pathways in ERM phosphorylation as well as their involvement in influenza virus-induced endothelial malfunction. The activation of p38MAPK, Rho/ROCK and PKC pathways upon influenza virus stimulation were observed, as evidenced by the evaluation of phosphorylated p38 (p-p38), phosphorylated MKK (p-MKK) in p38MAPK pathway, ROCK1 in Rho/ROCK pathway and phosphorylated PKC (p-PKC) in PKC pathway. We also showed that virus-induced ERM phosphorylation was reduced by using p38MAPK inhibitor, SB203580 (20 μM), Rho/ROCK inhibitor, Y27632 (20 μM), PKC inhibitor, LY317615 (10 μM). Additionally, influenza virus-induced F-actin reorganization and hyperpermeability were attenuated by pretreatment with SB203580, Y27632 and LY317615. Taken together, we provide the first evidence that p38MAPK, Rho/ROCK and PKC are involved in influenza-induced cytoskeletal changes and permeability increases in PMVEC via phosphorylating ERM.

A Chinese medicine granule, Shu-Feng-Xuan-Fei (SFXF), is critical for viral clearance in early phase of influenza virus infection. In this study, 72 ICR mice were randomly divided into six groups: normal control group, virus control group, Oseltamivir group, low-dose SFXF, medium-dose SFXF, and high-dose SFXF. Mice were anesthetized and inoculated with 4LD50 of influenza virus A (H1N1) except normal control group. Oseltamivir group received 11.375 mg·kg−1·d−1 Oseltamivir Phosphate. SFXF 3.76, 1.88 and 0.94 g·kg−1·d−1 were administrated to mice in all SFXF groups. Each group was in equal dose of 0.2ml daily for 4 consecutive days. Mice were sacrificed and then total RNA was extracted in lung tissue. Some genes involved in T-cell-mediated immunity were selected by DNA microarray. These candidate genes were verified by Real-Time PCR and western immunoblotting. Compared with virus control group, in Toll-like receptor signaling pathway, 12 virus-altered genes were significantly reduced following medium-dose SFXF treatment. Eighteen antigen processing presentation-associated genes were upregulated by medium-dose SFXF. In the process of T cell receptor signaling pathway, 19 genes were downregulated by medium-dose SFXF treatment. On exploration into effector T cells activation and cytokines, all of altered genes in virus control group were reversed by medium-dose SFXF. Real-time PCR and western immunoblotting showed that the regulation of medium-dose SFXF in IL-4, IFN-γ, TNF-α, IL-1β, TLR7, MyD88, p38, and JNK was superior to Oseltamivir and high-dose SFXF group. Therefore, SFXF granules could reduce influenza infected cells and activation of T cells.

OBJECTIVE:To investigate the expression of the inflammation-related cytokines in pneumonia mice infected with influenza virus and regulation of Shufengxuanfei(SFXF) and Jiebiaoqingli (JBQL) Chinese herbal anti-virus formulas.METHODS:Mice were anesthetized and then infected intranasally by dropping 0.05 mL of influenza virus suspension (4×LD50;) except normal group. Mice were divided randomly into nine groups: normal group, model group (virus only), control group [11.375 g/(kg.d)Oseltamivir], low-dose SFXF [0.94 g/(kg.d)], medium-dose SFXF [1.88 g/(kg.d)], high-dose SFXF [3.76 g/(kg.d)], low-dose JBQL [1.09 g/(kg.d)], medium-dose JBQL [2.18 g/(kg.d)] and high-dose JBQL [4.36 g/(kg.d)]. Oseltamivir group, SFXF groups and JBQL groups were administered to mice by oral gavage in equal dose of 0.2 mL daily for 4 consecutive days, while the rest of the groups received water only. Total RNA was extracted in each group. Then gene chips were used to screen these RNA samples. Select differentially expressed genes of cytokines involved in inflammation. Some candidate genes, such as IL-1β, IL-8, IL-10, RANTES and ICAM-1 were verified by qRT-PCR. To confirm the genes expression data from the microarray involved in inflammation in response to virus infection and treatment, we used qPCR to verify mRNA relative expressions of IL-1β, IL-8, IL-10, RANTES and ICAM-1. The expression of IL-1β protein in lung tissues was verified by Western blotting.RESULTS:IL-1β, CXCR2, CCL5, IL-10, IL-6, IL-18, TGF-β1 and CCL2 were up-regulated in model group. Gene expressions of IL-1β, CXCR2, CCL5, IL-10 and IL-6 were significantly down-regulated by all therapeutic groups. SFXF in medium-dose and low-dose down-regulated gene expressions of IL-18, TGF-β1, CCL2 and CCL5. IL-18 and CCL5 was down-regulated by both low-dose and medium-dose JBQL. qRT-PCR and western blot experiments showed that two formulas in medium-dose can down-regulate mRNA and protein expression of IL-1β (P<0.01). Both SFXF and JBQL in medium-dose significantly decreased the IL-8, RANTES, ICAM-1 and IL-10 mRNA expression (P<0.05 or P<0.01), compared with the model group. As expected, qRT-PCR data were in good agreement with the microarray assay.CONCLUSION:The two anti-viral formulas may inhibit inflammatory immunopathogenesis, and may have the actions of protection the lung tissue from influenza-induced injury.

Objective During viral infection,apoptosis by caspase pathway is induced as a consequence of activation of host cellular defense mechanism to limit the virus replication,but the mechanism of apoptosis infected with influenza virus remains to be clarified. The aim of this study was to investigate the related genes of apoptosis of pneumonia mice infected with influenza virus and the therapeutic effects of two herbal anti-virus formulas. Methods The model of pneumonia by nasal dropping influenza virus A was established in mice,and then divided randomly into nine groups: normal group,pneumonia model group,oseltamivir control group,low-dose,medium-dose and high-dose Shufengxuanfei groups,low-dose,medium-dose and high-dose Jiebiaoqingli groups. Total RNA was extracted in each group and gene chips were used to screen the genes involved in apoptosis. These candidate genes were verified by Real-Time PCR. Results Compared with the normal group,casp-8,casp-9,Fas,FasL,TNF,TNFrasf1a,Tradd,IL1a,IL1b,IL1r1,IL1r2 and casp-7gene increased in pneumonia model group. Casp-8,casp-7,Fas,FasL,TNF,TNFrasf1a,Tradd,IL1a,IL1b,IL1r1and IL1r2 gene decreased in medium-dose Shufengxuanfei group. The role of downregulation of these above genes in low dose Shufengxuanfei group was better than in low dose Jiebiaoqingli group. The therapeutic effects of medium dose Shufengxuanfei was better than in medium dose Jiebiaoqingli. Compared with the model group,the casp-3,-8,-9 mRNA expression decreased in low-dose Shufengxuanfei group and mediumdose Jiebiaoqingli group( P 0. 01). The therapeutic effects of different doses of Shufengxuanfei and Jiebiaoqingli as follows: low-dose Shufengxuanfei medium-dose Shufengxuanfei medium-dose Jiebiaoqingli low-dose Jiebiaoqingli. Conclusion The regulation effect of apoptosis gene in Shufengxuanfei formula was superior to that of Jiebiaoqingli. Shufengxuanfei formula suppressed casp-3,-8,-9 mRNA expression,and protected against the apoptosis induced by influenza virus.

OBJECTIVE To investigate the regulation of two herbal anti-virus formulas on gene expression profile associated with natural killer cell (NK cell) mediated cytotoxicity in pneumonia mice infected with influenza virus. METHODS According to random number table, 90 ICR mice were divided into nine groups with 10 mice in each group: normal group (N), model group (M), oseltamivir group (control group, C), low-dose, medium-dose and high-dose Shufeng Xuanfei formula groups (SL, SM, SH groups), and low-dose, medium-dose and high-dose Jiebiao Qingli formula groups (JL, JM, JH groups). The model of pneumonia was reproduced by nasal dropping influenza virus A (FM1) in mice. N group was given isotonic saline 0.05 ml in nasal drops. After 2 hours of model-building, C group was received 11.375 mg×kg⁻¹×d⁻¹ oseltamivir phosphate. Shufeng Xuanfei formula (mainly honeysuckle, forsythia and radix isatidis, etc.) with 3.76, 1.88 and 0.94 g×kg⁻¹×d were administrated to SH, SM and SL groups by gastric irrigation respectively. Jiebiao Qingli formula (mainly ephedra, gypsum, glycyrrhiza glabra, etc.) with 4.36, 2.18 and 1.09 g×kg⁻¹×d⁻¹ were administrated to JH, JM and JL groups by gastric irrigation respectively. In N and M groups, normal saline was administrated with gastric perfusion. Each group was in equal dose of 0.2 ml daily over a 4-day period. Total RNA in lung tissue of mice were extracted in each group, then gene chips were used to screen these RNA samples. Some genes involved NK cell mediated cytotoxicity were selected, with "I" representing of signal intensity. These candidate genes were verified by real-time fluorescent quantitation polymerase chain reaction (PCR) and Western blotting. RESULTS In the pathway of NK cell mediated cytotoxicity, M group up-regulated 43 genes expression, and 36, 29, 22, 21, 20 and 10 genes showed down-regulation in SM, JM, SL, JH, SH and JL groups, respectively. Apart from gene co-expression network in SH, SL, JH, JM and JL, SM also expressed other differential genes which SH, SL, JH, JM and JL did not. So medium-does Shufeng Xuanfei formula had the most significant regulation in gene expression of NK cell mediated cytotoxicity. By real-time PCR and Western blotting experiments showed that compared with the M group, mRNA and protein expression of tumor necrosis factor-α (TNF-α) in these two formula groups were significantly down-regulated, especially prominent in SM group and JM group (TNF-α mRNA: 1.07 ± 0.19, 1.19 ± 0.14 vs. 3.20 ± 0.56, both P<0.01). CONCLUSIONS Influenza viral replication in host cell, which means influenza antigens exposure in infected cells as target cells. NK cells recognize and exert cell mediated cytotoxic function against influenza antigens. Genes associated with NK cell mediated cytotoxicity in influenza infection were up-regulated. Shufeng Xuanfei and Jiebiao Qingli formulas could down-regulate these genes. The mechanism of down-regulated genes is that the number of influenza infected cells and NK cells activation decreases in treatment with two formulas.

目的研究黄芩苷对甲型流感病毒性肺炎小鼠肺组织炎性损伤的作用机制。方法将96只ICR鼠随机分为正常对照组、模型组、利巴韦林组（100 mg.kg-1）,黄芩苷组（1 500、750、375 mg.kg-1）[5],每组16只。用流感病毒亚甲型鼠肺适应株A/FM/1/47（H1N1）感染小鼠,制备小鼠流感病毒性肺炎模型,用不同剂量黄芩苷溶液灌胃治疗后,观察肺组织H-E染色切片病理变化,采用RT-PCR测定肺组织c-jun、c-fos mRNA表达,采用Western blot测定肺组织c-jun、磷酸化c-jun蛋白表达,应用ELISA检测肺匀浆炎性细胞因子TNF-α、IL-1β的含量。结果黄芩苷750、375 mg.kg-1剂量均能明显减轻肺组织炎性损伤;明显降低c-jun、c-fosmRNA的表达（P＜0.05,P＜0.01）,明显降低c-jun、磷酸化c-jun蛋白表达（P＜0.01）;明显抑制TNF-α、IL-1β的分泌（P＜0.01）。结论黄芩苷能够缓解甲型流感病毒诱导的炎性病理损伤,其可能通过抑制流感病毒感染引起的转录因子AP-1高表达而降低炎性细胞因子的分泌水平,从而发挥抗流感病毒感染的作用。

OBJECTIVE:To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo. METHODS:In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase assays in Madin Darby canine kidney cells. In vivo anti-influenza virus assays were performed on the viral pneumonia model of mice. The numbers of mice that died within day 2 to day 14 postinfection were recorded to calculate the mortality. On days 2, 4, and 6, the viral titers in the lungs were determined by hemagglutination assay; hematoxylin/eosin staining was used to assess the pathogenic changes of lung tissues; the concentrations of tumor necrosis factor-alpha (TNF-α) and monocyte specific chemoattractant molecule (MCP-1) were measured by radio immunoassay or enzyme-linked immunosorbent assay; the concentrations of nitric oxide (NO) and inducible nitric oxide synthetase (iNOS) were detected by colorimetric method; reverse transcription polymerase chain reaction was used to detect the mRNA level of TNF-α and MCP-1. RESULTS:Berberine showed inhibitory effects on cytopathogenic effects and neuraminidase activity of virus, with the therapeutic index 9.69. In vivo, berberine decreased mice mortality from 90% to 55%, reduced virus titers in the lungs on day 2 postinfection (P<0.05). The lung histology scores were 1.50 ± 0.67, 4.50 ± 1.00, and 5.50 ± 1.00 in the berberine group on days 2, 4, and 6, respectively, which were significantly reduced compared to 2.17 ± 0.22, 6.83 ± 0.44, and 8.50 ± 0.33 in the infected group (P<0.05). The productions of NO and iNOS were repressed by berberine compared with those in the infected group (P<0.01). The transcription and expression of TNF-α were inhibited by berberine on day 4 (P<0.01) and day 6 (P<0.05), and those of MCP-1 were inhibited on day 6 (P<0.01) compared with the infected group. CONCLUSIONS:Berberine exhibited antiviral effects on the influenza virus both in vitro and in vivo. The possible therapeutic mechanism of berberine on influenza-induced viral pneumonia might be inhibiting the virus infection, as well as improving the pathogenic changes by repressing inflammatory substances release.

OBJECTIVE:To observe the effects of ginsenoside (Gs) and berberine (Ber), two kinds of active components of traditional Chinese herbal medicine, on transforming growth factor-beta1 (TGF-beta1) and prostaglandin E2 (PGE2) in PG cells.METHODS:Co-culture system of human lung carcinoma cell line PG and human T lymphocyte cell line Jurkat was established. PG cells were treated with Gs (100 microg/ml) and Ber (10 mug/ml) for twenty-four hours, and then cocultured with Jurkat cells. After 24-hour coculture, the state of Jurkat cells was observed with inverted microscope. The viable count of Jurkat cells was detected by trypan blue staining after 6- and 24-hour coculture, and the apoptosis of Jurkat cells was evaluated by flow cytometry. PG cells were treated with 100, 50, 25 microg/ml Gs and 10, 5, 2.5 microg/ml Ber respectively, and the content of TGF-beta1 and PGE(2) in PG cells was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA) method.RESULTS:After coculture with PG cells treated with Gs and Ber, the number of Jurkat cells was less than blank control group, and the apoptosis rates of Jurkat cells in Gs- and Ber-treated groups were higher than blank control group. Gs and Ber could promote the secretion of TGF-beta1 in PG cells, but could not change the level of PGE(2).CONCLUSION:Gs and Ber can promote the growth inhibition and apoptosis of Jurkat cells induced by PG cells, which may be related to the up-regulation of Gs and Ber on TGF-beta1 secretion in PG cells.