Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW...Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.展开更多
While our understanding of male reproductive strategies is informed by extensive investigations into endocrine mechanisms, the proximate mechanisms by which females compete for mates and adjust reproduction to social ...While our understanding of male reproductive strategies is informed by extensive investigations into endocrine mechanisms, the proximate mechanisms by which females compete for mates and adjust reproduction to social environment remains enigmatic. We set out to uncover endocrine correlates of mate choice, social environment, and reproductive investment in female red-backed fairy-wrens Malurus melanocephalus. In this socially monogamous, yet highly sexually promiscuous species, females experience discrete variation in the phenotype of their mates, which vary in both plumage signals and level of paternal care, and in the composition of their breeding groups, which consist of either the pair alone or with an additional cooperative auxiliary; fe- male investment varies according to these social parameters. We found that androgen, estrogen, and glucorticoid levels varied with reproductive stage, with highest androgen and estrogen concentrations during nest construction and highest corticosterone concentrations during the pre-breeding stage. These stage-dependent patterns did not vary with male phenotype or auxiliary presence, though androgen levels during pre-breeding mate selection were lower in females obtaining red/black mates than those obtaining brown mates. We found no evidence that androgen, estrogen, or corticosterone levels during the fertile period were re- lated to extra-pair young （EPY） frequency. This study demonstrates clear changes in steroid levels with reproductive stage, though it found little support for variation with social environment. We suggest hormonal responsiveness to social factors may be physiologically constrained in ways that are bypassed through exogenous hormone manipulations.展开更多
文摘Objective: To investigate the role of PKC isoforms in the regulation of LPS-triggered tumoricidal activity in macrophages and further elucidate its signal mechanisms. Methods: Two macrophage cell lines (P388D1 and RAW264.7) were stimulated by LPS alone, or with long-term of PMA pretreatment. Then cytotoxicities to P815 cells (by MTT assay) and IL-1, TNF- (by ELISA) and nitric oxide (NO) production (by Griess reagent) in supernatants were measured. Western blot for PKC isoforms after long-term PMA pretreatment was analyzed. Results: RAW264.7 cells were stimulated with LPS to kill target tumor cells P815, whereas P388D1 cells failed to develop such an ability. Down-regulation of PKC isoforms by chronic treatment with PMA significantly inhibited the LPS-induced cytotoxicity in RAW264.7 cells. In unstimulated state, Western blotting with rabbit antiserum specific for the PKC, 1, 2, or showed all 5 isoforms were detected in P388D1 cells, while only PKC, PKC1 and PKC were detected in RAW264.7 cells. Exposure of the cells to long-term of PMA treatment significantly down-regulated the expression of PKC, PKC1 and PKC in RAW264.7 cells. But in P388D1 cells, although PKC, PKC and PKC were down-regulated, the expression of PKC1 and PKC2 could not be regulated. Comparing with LPS-induced IL-1, TNF- and NO production by the two macrophage cell lines, P388D1 failed to produce NO. In RAW264.7 cells, LPS-induced NO production and antitumor activity was attenuated by the addition of L-NAME, an iNOS inhibitor. Conclusion: The results indicated a critical role of PKC in LPS-induced antitumor activity and this cytotoxicity is mainly due to PKC- mediated NO production by RAW264.7 cells, but not a direct cytotoxic activity.
基金Acknowledgement We sincerely appreciate the commendable field efforts of a large number of field technicians who assisted with data collection during the course of this study, as well as logistical support provided by B. Congdon, T. Daniel, J. Lindsay and D. Westcott. We also thank members of the Schwabl and Webster labs for their valuable input throughout. Thanks also to Becca Sail＇an and Maren Vitousek for the invitation to contribute to this volume. This research was conducted with appropriate permits and permissions from the governments of Queensland and Australia, and material is based upon work supported by the National Science Foundation （USA） through grants to MSW and HS and a graduate traineeship to DGB.
文摘While our understanding of male reproductive strategies is informed by extensive investigations into endocrine mechanisms, the proximate mechanisms by which females compete for mates and adjust reproduction to social environment remains enigmatic. We set out to uncover endocrine correlates of mate choice, social environment, and reproductive investment in female red-backed fairy-wrens Malurus melanocephalus. In this socially monogamous, yet highly sexually promiscuous species, females experience discrete variation in the phenotype of their mates, which vary in both plumage signals and level of paternal care, and in the composition of their breeding groups, which consist of either the pair alone or with an additional cooperative auxiliary; fe- male investment varies according to these social parameters. We found that androgen, estrogen, and glucorticoid levels varied with reproductive stage, with highest androgen and estrogen concentrations during nest construction and highest corticosterone concentrations during the pre-breeding stage. These stage-dependent patterns did not vary with male phenotype or auxiliary presence, though androgen levels during pre-breeding mate selection were lower in females obtaining red/black mates than those obtaining brown mates. We found no evidence that androgen, estrogen, or corticosterone levels during the fertile period were re- lated to extra-pair young （EPY） frequency. This study demonstrates clear changes in steroid levels with reproductive stage, though it found little support for variation with social environment. We suggest hormonal responsiveness to social factors may be physiologically constrained in ways that are bypassed through exogenous hormone manipulations.
