Ginkgetin alleviates LPS-induced inflammatory damage in macrophages of mouse model via inhibiting IκB-α degradation / 中国药理学通报

Aim To explore the anti-inflammatory effect of natural compound Ginkgetin on lipopolysaccharide-induced mouse primary peritoneal macrophages and the underlying mechanism, in order to provide a theoretical basis for the development of clinical drug candidates. Methods MTT test kit was used to detect the cytotoxicity of Ginkgetin on mouse primary peritoneal macrophages; ELISA and RT-qPCR methods were used to detect the anti-inflammatory effect of different concentrations of Ginkgetin on LPS-induced cell inflammation injury model; Western blot was used to detect the anti-inflammatory mechanism of ginkgo flavonoids. Results Compared with LPS stimulation group, Ginkgetin treatment group produced a concentration-dependent anti-inflammatory effect, which could be attributed to the fact that Ginkgetin could inhibit LPS-induced activation of NF-κB signaling pathway. MTT results also showed that ginkgo flavonoids had little toxicity to mouse primary peritoneal macrophages. Conclusions Ginkgelin alleviates LPS-induced inflammatory injury of mouse primary peritoneal macrophages by inhibiting the activation of NF-κB signaling pathway. It is expected to be a natural monomer antiinflammatory drug candidate.

Effect and regulatory mechanism of peritoneal macrophage polarization in the progress of acute pancreatitis / 中华消化外科杂志

Acute pancreatitis (AP) is a local and systemic inflammation process which is characterized by extensive infiltration of inflammatory cells and outbreak of inflammatory factors.Recent researches have shown that macrophage play a critical role in the process of AP,in particular,peritoneal macro phage as a main innate immune cell in the peritoneal cavity which has been reported not only serve as the first line in host defense,but also help maintaining the immune homeostasis and coordinate the repair and regenerate response by regulating the polarization of their phenotype.Recently,peritoneal macrophages could interact with ascetic fluid of AP patient,and further affect the development and resolution of inflammation.In this review,authors focused on the role and regulatory mechanism of peritoneal macrophage in the progress of AP.

Effect of Porphyromonas gingivalis lipopolysaccharide on gene expression of inflammatory cytokines in peritoneal macrophages of hyperlipidemic rabbits / 口腔疾病防治

Objective @#To compare the inflammatory reaction of peritoneal macrophage after Pg-LPS stimulated in healthy rabbit and hyperlipidemia rabbit. @*Methods @#12 New Zealand rabbits were randomly divided into 2 groups with 6 rabbits in each group, and normal diet and high-fat diet were fed to them respectively. The hyperlipidemia model was set up after 6 weeks. The peritoneal macrophage in normal and hyperlipidemia group were isolated and cultured, and then the cells in both groups were respectively divided into 3 groups: control, 1 μg/mL Pg-LPS, and 1 μg/mL E.coli-LPS. After 24 h treatment, the expressions of C-reaction protein (CRP), interleukin-1β (IL-1β), interleukin-6 (IL-6) , interleukin-8 (IL-8), and tumor necrosis factor-α (TNF-α) were detected by realtime PCR. @*Results @#The levels of CRP, IL-1β, IL-6, IL-8, and TNF-α were higher in hyperlipidemia control group than normal control group. The expressions of inflammatory substances were increased after stimulated by Pg-LPS, and statistically higher in hyperlipidemia rabbit group than normal group (P < 0.05). @*Conclusion@#Pg-LPS can enhance the mRNA expressions of inflammation related factors in peritoneal macrophage in hyperlipidemia rabbit.

In vitro Staphylococcus aureus-induced oxidative stress in mice murine peritoneal macrophages：a duration-dependent approach

Objective: To evaluate the free radical generation and status of the antioxidant enzymes in murine peritoneal macrophage during in vitro vancomycin sensitive Staphylococcus aureus (VSSA) treatment with different time interval.Methods:Peritoneal macrophages were treated with 5í106 CFU/mL VSSA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity, nitric oxide generation, antioxidant enzyme status and components of glutathione cycle were analyzed.Results:Superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity and nitric oxide generation got peak at 3 h, indicating maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during VSSA infection. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-s-transferase activity were decreased significantly (P<0.05) with increasing time of VSSA infection. But the oxidized glutathione level was time dependently increased significantly (P<0.05) in murine peritoneal macrophages. All the changes in peritoneal macrophages after 3 h in vitro VSSA treatment had no significant difference.Conclusions:From this study, it may be summarized that in vitro VSSA infection not only generates excess free radical but also affects the antioxidant status and glutathione cycle in murine peritoneal macrophages.

In vitro Staphylococcus aureus-induced oxidative stress in mice murine peritoneal macrophages: a duration-dependent approach

<p><b>OBJECTIVE</b>To evaluate the free radical generation and status of the antioxidant enzymes in murine peritoneal macrophage during in vitro vancomycin sensitive Staphylococcus aureus (VSSA) treatment with different time interval.</p><p><b>METHODS</b>Peritoneal macrophages were treated with 5×10(6) CFU/mL VSSA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity, nitric oxide generation, antioxidant enzyme status and components of glutathione cycle were analyzed.</p><p><b>RESULTS</b>Superoxide anion generation, NADPH oxidase activity, myeloperoxidase activity and nitric oxide generation got peak at 3 h, indicating maximum free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during VSSA infection. Reduced glutathione level, glutathione peroxidase, glutathione reductase, and glutathione-s-transferase activity were decreased significantly (P<0.05) with increasing time of VSSA infection. But the oxidized glutathione level was time dependently increased significantly (P<0.05) in murine peritoneal macrophages. All the changes in peritoneal macrophages after 3 h in vitro VSSA treatment had no significant difference.</p><p><b>CONCLUSIONS</b>From this study, it may be summarized that in vitro VSSA infection not only generates excess free radical but also affects the antioxidant status and glutathione cycle in murine peritoneal macrophages.</p>

IL-33 Priming Enhances Peritoneal Macrophage Activity in Response to Candida albicans

IL-33 is a member of the IL-1 cytokine family and plays a role in the host defense against bacteria, viruses, and fungi. In this study, we investigated the function of IL-33 and its receptor in in vitro macrophage responses to Candida albicans. Our results demonstrate that pre-sensitization of isolated peritoneal macrophages with IL-33 enhanced their pro-inflammatory cytokine production and phagocytic activity in response to C. albicans. These macrophage activities were entirely dependent on the ST2-MyD88 signaling pathway. In addition, pre-sensitization with IL-33 also increased ROS production and the subsequent killing ability of macrophages following C. albicans challenge. These results indicate that IL-33 may increase anti-fungal activity against Candida through macrophage-mediated resistance mechanisms.

Effects of serum containing Dahuang Fuzi Decoction on JAK2/STAT3 signal pathway in mice with severe acute pancreatitis / 中草药

Objective: To study the effects of serum containing Dahuang Fuzi (Rhubarb and Aconite) Decoction (DFD) on JAK2/STAT3 signal pathway in mice with severe acute pancreatitis (SAP). Methods: SAP model in mice was constructed, and then the peritoneal macrophages were vaccinated into the culture plate to set model group, serum containing DFD groups (2.5%, 5%, and 10%), AG490 (10 μmol/L ) positive group, and peritoneal macrophages of normal mice acted as normal control group. After an incubation of 2 h, cells were added with serum containing DFD at different concentration or AG490 0.5 mL, 24 h later, the concentration of TNF-α and IL-6 in supernatant were determined by quantitative sandwich enzyme-linked immunosorbent assay (ELISA) kits, mRNA and protein expression levels of JAK2 and STAT3 in cells were evaluated by Western blotting. Results: DFD could significantly decrease the levels of cytokines TNF-α and IL-6 in the supernatants, inhibit the mRNA and protein expression levels of JAK2 and STAT3 in peritoneal macrophages of SAP mice. Conclusion: DFD could inhibit the JAK2/STAT3 signal pathway and inflammatory responses in peritoneal macrophages of SAP mice.

Susceptibility of peritoneal macrophage from different species of neotropical primates to Ex vivo Leishmania (L.) infantum chagasi-infection / Susceptibilidade do macrófago peritoneal de diferentes espécies de primatas neotropicais para a infecção ex vivo por Leishmania (L.) infantum chagasi

This study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-α, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-a response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mL), C. penicillata (154/130 pg/mL), S. sciureus (164/104 pg/mL) and A. azarae infulatus (154/104 pg/mL), with exception of C. goeldii (38/83 pg/mL). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.

Este estudo examinou a susceptibilidade do macrófago peritoneal (PM) dos primatas neotropicais: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus e Callimico goeldii para a infecção ex vivo por Leishmania (L.) infantum chagasi, o agente etiológico da leishmaniose visceral americana (LVA), como método de triagem para avaliar o potencial desses primatas como modelo de estudo da LVA. A susceptibilidade do PM para a infecção foi investigada através do índice de infecção do PM (PMI) a intervalos de 24, 72 horas e, ainda, pela média dessas taxas (FPMI), assim como, pelas respostas do TNF-α, IL-2 (ELISA de captura) e óxido nítrico (NO) (método de Griess). Às 24hs da infecção experimental, o PMI do primata A. azarae infulatus (128) foi maior que aqueles de C. penicillata (83), C. goeldii (78), S. sciureus (77) e C. jacchus (55). Às 72hs, houve uma redução significativa do PMI de quatro primatas: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) e C. jacchus (55/12), com exceção de C. goeldii (78/54). O FPMI dos primatas A. azarae infulatus (82.5) e C. goeldii (66) foi maior que do primata C. jacchus (33.5), porém, não foi maior que dos primatas C. penicillata (60.5) e S. sciureus (57.5). A resposta do TNF-α foi mais regular nos quatro primatas que reduziram o PMI no intervalo de 24-72hs: C. jacchus (145/122 pg/µL), C. penicillata (154/130 pg/µL), S. sciureus (164/104 pg/µL) e A. azarae infulatus (154/104 pg/µL), com exceção de C. goeldii (38/83 pg/µL). A resposta de IL-12 foi, principalmente, marcante nos primatas A. azarae infulatus e C. goeldii, os quais apresentaram as maiores taxas do FPMI, e a resposta do NO foi maior no primata C. goeldii, em especial no intervalo de 72hs. Estes achados sugerem, fortemente, que estes primatas neotropicais parecem ter desenvolvido mecanismos resistentes de resposta imune inata capaz de controlar o crescimento intracelular da infecção por L. (L.) i. chagasi no macrófago, o que não encoraja o uso destes primatas como modelo de estudo da LVA.

Alteration of some cellular function in amikacin resistant Pseudomonas aeruginosa transfected macrophages：a time dependent approach

To evaluate the free radical generation and antioxidant enzymes status in murine peritoneal macrophage during in vitro amikacin resistant Pseudomonas aeruginosa (ARPA) treatment with different time interval. Methods: Peritoneal macrophages were treated with 1×108 CFU/mL ARPA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and super oxide anion generation, NO generation, reduced glutathione level and antioxidant enzymes status were analyzed. Results: Super oxide anion generation and NO generation got peak at 12 h, indicating maximal free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during ARPA transfection. Reduced glutathione level and antioxidant enzymes status were decreased significantly (P<0.05) with increasing time of ARPA transfection. All the changes in peritoneal macrophages after 12 h in vitro ARPA transfection had significant difference (P<0.05). Conclusions: From this study, it may be summarized that in vitro ARPA infection not only generates excess free radical but also affects the antioxidant system and glutathione cycle in murine peritoneal macrophage.

Alteration of some cellular function in amikacin resistant Pseudomonas aeruginosa transfected macrophages: a time dependent approach

<p><b>OBJECTIVE</b>To evaluate the free radical generation and antioxidant enzymes status in murine peritoneal macrophage during in vitro amikacin resistant Pseudomonas aeruginosa (ARPA) treatment with different time interval.</p><p><b>METHODS</b>Peritoneal macrophages were treated with 1×10(8) CFU/mL ARPA cell suspension in vitro for different time interval (1, 2, 3, 6, 12, and 24 h) and super oxide anion generation, NO generation, reduced glutathione level and antioxidant enzymes status were analyzed.</p><p><b>RESULTS</b>Super oxide anion generation and NO generation got peak at 12 h, indicating maximal free radical generation through activation of NADPH oxidase in murine peritoneal macrophages during ARPA transfection. Reduced glutathione level and antioxidant enzymes status were decreased significantly (P<0.05) with increasing time of ARPA transfection. All the changes in peritoneal macrophages after 12 h in vitro ARPA transfection had significant difference (P<0.05).</p><p><b>CONCLUSIONS</b>From this study, it may be summarized that in vitro ARPA infection not only generates excess free radical but also affects the antioxidant system and glutathione cycle in murine peritoneal macrophage.</p>

Preparation of lipsomal Clodronate and its inducing effect on apoptosis of peritoneal macrophages in rats after severe acute pancreatitis / 中华肝胆外科杂志

Objective To explore the preparation of liposomal clodronate and investigate its inducing effects on the apoptosis of peritoneal macrophages in rats after severe acute pancreatitis (SAP).Methods Liposomal clodronate was prepared by means of thin film. SAP rat model was established by retrograde injection of 5% sodium taurocholate into the pancreatic duct. The peritoneal macrophages were obtained from SAP rats. After exposure to different doses of liposomal clodronate (50, 100,150 μl), the PM proliferation was determined by MTT colourimetry. The apoptosis of PM was measured by flow cytometry and agarose gel electrophoresis, respectively. Results The prepared liposomal clodronate had a suitable encapsulation efficiency of clodronate (5.8%) with an average size of 200 nm. The spherical shape of liposome was confirmed by transmission electron microscope. Exposed to liposomal clodronate of different doses resulted in a obvious growth depression (P＜0.01). The apoptotic rate of the PM was (10.32±0.34) %, (18.16±0.49)% and (29.87±0.35)% in three different dose groups and the difference was marked (P＜0.01). 1.2% of agarose gel electrophoresis of DNA extracted from apoptotic macrophages induced by liposomal clodronate showed clearer and characteristic ladder following the liposomal clodronate concentration. Conclusion Liposomal clodronate has a definite effect on peritoneal macrophages in SAP rats.

The photological function of MPA coated CdTe QDs and their biocompatibility / 细胞与分子免疫学杂志

AIM: To investigate the CdTe quantum dots coated with MPA and explore its biocompatibility with living cells. METHODS: CdTe quantum dots coated with MPA were prepared in aqueous phase and MPA CdTe QDs were Characterized with TEM, fluorospectrophotometer and ultraviolet spectrophotometer. QDs were Modified with with avidin, purified and prepared as flurescent probe. LSCM was used to observe the expression of MHC Ⅱ antigen on PMφ cells, which was labeled by QDs. Cell culture and MTT assays were used to determine the biocompatibility of MPA coated CdTe quantum dots with the B-16 cells as target cells. RESULTS: The particle diameter of CdTe quantum dots prepared in aqueous phase was well distributed. They had good photological performance and greater stability after coated with MPA. MHC Ⅱ antigen on PMφ was labeled with the QDs-Avidin fluorescent probe showed great fluorescence intensity, which was easy to be detected by fluorescence microscope and LSCM. MPA CdTe QDs showed cytotoxicity when its density was very high, but they showed little cytotoxicity during the normal use of influence label density limit. CONCLUSION: MPA CdTe QDs can be used as new fluorescent lable as they are of even size, not easy to bleach or quench, have good photological performance and stability and good biocompatibility.

Mechanisms of phagocytosis of Leptospira by peritoneal macrophages of guinea pigs / 上海交通大学学报（医学版）

Objective To investigate the mechanisms of phagocytosis of virulent Leptospira by peritoneal macrophages of guinea pigs,andevaluatetheroleof innateimmuneinthepathogenesisof leptospirosis. Methods Peritoneal macrophages of guinea pigs were extracted. Three specific inhibitors ( microfilament inhibitor cytochalasin D,microtube inhibitor colchicine and PI3K signalling pathway inhibitor LY294002) were added respectively to the macrophages 1 h before the infection of virulent Leptospira interrogans serovar Lai type strain Lai in vitro.Meanwhile, control group without inhibitor was established.Phagocytosis was observed by laser scanning confocal microscopy and phagocytic rates were evaluated by flow cytometry 3 h after infection.ResultsThe phagocytic rates of control group, cytochalasin D group, colchicine group and LY294002 group were (38.98 ± 0.91)%,(23. 99 ± 1. 40) % ,(40.81±0.91)% and (39.64 ±3.56) %, respectively.The phagocytic rate of cytochalasin D group was significantly lower than that of control group (P < 0. 05), while those of colchicine group and LY294002 group were not significantly different from that of control group (P >0.05). ConclusionMicrofilaments play an important role in the phagocytosis of strain Lai by peritoneal macrophages,but the process is independent on PI3K signalling pathway,and microtubes play little part during the phagocytosis.

Regulation effect of taurochenodeoxycholic acid immunologic function in mice / 中国药理学通报

Aim To study the effect of Taurochenodeoxycholic Acid (TCDCA) on immunologic cell in vitro and in vivo in mice. Methods Five doses were used , including 0.043, 0.065, 0.1, 0.153, 0.189 g?kg-1 in vivo and 0.01, 0.05, 0.1, 1, 10 mg?L-1 in vitro. Effects of TCDCA on peritoneal macrophage in mouse in vitro,splenic lymphocyte, normal organism, immunologic functional inhibiting model and nerve-endocrine-immunoregulation system were investigated. Results TCDCA had the obvious immunoregulation effect on normal organisms in mouse. 0.1 g?kg-1 TCDCA had the obvious anastated effects on the immunological inhibiting model made up by the cyclosporine A and cyclophosphamide. TCDCA may affect the peritoneal macrophage and splenic lymphocyte in vitro directly, meanwhile two kinds of immunological cells showed the obvious immunoregulation effects on the whole. 0.1 g?kg-1 TCDCA can increase the hydrocortisone contents of the nerve-endocrine-immunoregulation system in mouse most remarkably. Conclusion TCDCA had the adjustment effect on the immunoregulation function in mouse.

Effects of the composition of peritoneal dialysis solution on phagocytic function of peritoneal macrophage in rats / 대한내과학회지

BACKGROUND: Continuous ambulatory peritoneal dialysis (CAPD) has been accepted as one of the long-term treatment modalities for end-stage renal disease and peritonitis is the most important complication in CAPD therapy and first-line defense activity was achieved by peritoneal macrophage(PM). Our study has compared the phagocytic activity in rat PM according to various characteristic CAPD solutions(pH, glucose, Ca++, and erythropoietin). METHODS: Peritoneal cells were isolated from Sprague-Dawley rat with RPMI(4C(degrees), 100 ml). Various characteristic solutions were prepared. The solutions contained 1.5% or 4.25% dextrous, pH 5.4 or pH adjustment to 7.4, low calcium concentration or high calcium concentration and free erythropoietin or 2000U/L erythropoietin. Phagocytic activity of PM was measured by Candida albicans(ATCC 10231) under the inverted microscope. RESULTS: There was significantly increased phagocytic activity of rat PM in pH 7.4 solution than in pH 5.4 solution(61.8% vs. 72.4% in 1.5% dextrous solution; p 0.05, 61.6% vs. 45.3% in 4.25% dextrous solution; p 0.05, 61.6% vs. 52.3% in 4.25% dextrous solution; p< 0.05) and high glucose concentration affected to phagocytic activity in 2000U/L erythropoietin solution. CONCLUSIONS: These findings suggest that phagocytic activity of rat PM was increased in pH 7.4 solution and decreased in low calcium concentration and solution which was added erythropoietin, especially in high glucose solution. But glucose concentration did not affect phagocytic activity of rat PM independently.

Comparison of Macrophage Activation and Tumor - cytotoxicity in Mouse and hamster Peritoneal Macrophages by Cold Stress / 대한면역학회지

No abstract available.

Effect of Mercury Chloride on Peritoneal Macrophage or EMT-6 cell from Balb/c mice / 대한산업의학회지

The effect of treatment of mercury chloride on the nitrite and nitrate synthesis was observed in peritoneal macrophages from Balb/c mice and EMT-6 cells in vitro. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with cytokines. Amounts of nitrite and nitrate in the culture media after 24 and 36 hours of culture were about 2-fold, and 3-fold of those measured after 12 hours respectively. There were very close associations between the amounts of nitrite and nitrate measured in the culture media, according to culture time. The survival rate of peritoneal macrophages was significantly decreased by mercury chloride added into the media in dose-dependent manner, however the survivals of EMT-6 cells were not influenced by mercury chloride concentration in media. Nitrite and nitrate syntheses were dose-dependently decreased by mercury chloride added in culture media. These results reported here suggest that the disorder of cell mediated immunity by mercurials could be related to the inhibition of nitric oxide synthesis which seems to be caused by the inhibition of metabolism of cells.

Effect of Mercury Chloride on Peritoneal Macrophage or EMT-6 cell from Balb/c mice / 대한산업의학회지

The effect of treatment of mercury chloride on the nitrite and nitrate synthesis was observed in peritoneal macrophages from Balb/c mice and EMT-6 cells in vitro. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with cytokines. Amounts of nitrite and nitrate in the culture media after 24 and 36 hours of culture were about 2-fold, and 3-fold of those measured after 12 hours respectively. There were very close associations between the amounts of nitrite and nitrate measured in the culture media, according to culture time. The survival rate of peritoneal macrophages was significantly decreased by mercury chloride added into the media in dose-dependent manner, however the survivals of EMT-6 cells were not influenced by mercury chloride concentration in media. Nitrite and nitrate syntheses were dose-dependently decreased by mercury chloride added in culture media. These results reported here suggest that the disorder of cell mediated immunity by mercurials could be related to the inhibition of nitric oxide synthesis which seems to be caused by the inhibition of metabolism of cells.

Effects of Nitrogen Oxidation Metabolism of Murine Macrophages on the Growth of MBT-2 Cell Line / 대한비뇨기과학회지

Recently it is known that nitric oxide(NO) generated by an activatedmacrophage plays an important role in tumoricidal or bactericidal activity. Thisstudy was done to know the effects of NO produced by the activated macrophages onthe growth of the murine bladder tumor cell line (MBT-2). For the activation ofmacrophages, RAW 264.7 cell line ( macrophage-derived cell line) and peritonealmacrophages from Balb/c mouse were treated with interferon-r (INF-r),lipopolysaccharide ( LPS) or INF-r plus LPS and for the evaluation of growthinhibition of MBT -2 cell line, tritiated thymidine (3[H] -thymidine)incorporation was measured after coculturing of MBT-2 cell line with macrophageswhich had been activated to produce NO. The results are as follows : l.Peritoneal macrophages from Balb/c mouse could be activated to produce NO onlywhen they are stimulated by the combination of INF-r and LPS (35+/-1.0uM/L). Theycould produce a slight increase amount of NO when they had been stimulated byINF-r (11+/-2.5uM/ L) or LPS (14+/-3.5uM/L) compared to control macrophages(8+/-2.5uM/L). The induction of NO production by INF-r plus LPS could be abrogatedby the use of NG-monomethyl-L- arginine (NGMMA), a competitive inhibitor of NOsynthase (5+/-1.5 M/L). 2. RAW 264.7 cell lines could be activated to produce NOby the treatment of INF-r, LPS, and INF-r plus LPS (40+/-2.5uM/L, 37+/-3.0uM/Land 51+/-2.6uM/L respectively). When they are treated with NGMMA, they produced nomore NO even though they had been stimulated with INF-r plus LPS (14+/-4.0uM/L),and they could produce small .amount of NO(19+/-1.0uM/L) in the absence of thestimulation. 3. The incorporation of 3[H] -thymidine of the MBT-2 and peritonealmacrophages was reduced when the cells had been treated with INF-r plus LPScompared to the control (14,519+/-1,087cpm, 20,716+/-1,474cpm respectively).There was no reduction in the incorporation of 3[H] -thymidine when the cellshad been treated with INF-r or LPS alone. The incorporation of 3[H]-thymidineincreased when the cells had been treated with INF-r plus LPS in the presence ofNGMMA(19,622+/-1,341cpm). 4. The incorporation of 3[H] -thymidine of the MBT-2 and RAW 264.7 cell lines were reduced when the cells had been treated with INF-r, LPS and INF-r plus LPS (19.068+/-144cpm, 15,070+/-122cpm and 7,543+/-85cpm respectively) compared to control( 20,708+/-142cpm). When the cells had been pretreated with NGMMA, the incorporation of 3[H]- thymidine recovered to same degree (12,605+/-108cpm) compared to the cells stimulated with INF-r plus LPS. The above correlation of the NO production from macrophages which had been stimulated by INF-r and/or LPS and the inhibition of growth of the tumor cells suggests that NO produced by stimulated macrophages might be the responsible molecule in the defense system of the body against tumors.

COMPARATVE STUDY OF QUANTITATIVE CYTOCHEMISTRY ON THE BCG ACTIVATED ALVEOLAR MACROPHAGES WITH THAT OF PERITONEAL MACROPHAGES / 解剖学报

Some intracellular enzyme-activities of BCG-activated alveolar macrophages (AM) and peritoneal macrophages (PM) were studied. It was found that (in normal) the activities of acid phosphatas (ACP), succinate dehydrogenase (SDH), non-specific esterase (NSE) in AM were higher than those in PM, but that of lactate dehydrogenase (LDH) was lower. Both in BCG activated AM and PM, the activities of ACP, SDH, NSE increased significantly, and that of LDH also enhanced in activated AM but not in PM.