Using inducers to induce cells to produce inflammatory response is a common in vitro experimental method to study inflammation.However,there are relatively few inflammatory models developed for the cosmetic industry,a...Using inducers to induce cells to produce inflammatory response is a common in vitro experimental method to study inflammation.However,there are relatively few inflammatory models developed for the cosmetic industry,and there are also great differences in the control of model induction,the selection of inflammatory indicators,and the concentration of inducers.Therefore,in this paper,by systematically studying the effects of Lipopolysaccharide(LPS)on the cell viability,the levels of IL-1α,IL-8 and ROS of human primary keratinocytes,a skin inflammation model based human primary keratinocyte was developed.The results showed that 0.01~100μg/mL LPS had no significant effect on the cell viability of human primary keratinocytes,while 100μg/mL LPS could simultaneously induce human primary keratinocytes to produce large amounts of IL-1αand IL-8,and 0.01μg/mL LPS could induce plentiful ROS.Therefore,a skin inflammation model for differential induction of different inflammatory indicators was established,and the sample OSM2021041301 was tested with this model,it was found that sample OSM2021041301 could significantly inhibit LPS-induced elevated IL-1αand IL-8 levels,the inhibitory effect on LPS-induced elevated ROS level was weak.The results indicated that OSM2021041301 has certain anti-inflammatory effect on inflammation caused by the increase of IL-1α,IL-8 and ROS induced by LPS and its analogues.展开更多
目的：人乳头瘤病毒16型（human papillomavirus，HPV 16）与包括宫颈癌在内的多种肿瘤的发生明确相关。诱导宿主基因组不稳定性可能是HPV16致瘤的重要机制之一。本研究对HPV16型E6蛋白诱导人原代角质形成细胞（primary human keratinoc...目的：人乳头瘤病毒16型（human papillomavirus，HPV 16）与包括宫颈癌在内的多种肿瘤的发生明确相关。诱导宿主基因组不稳定性可能是HPV16致瘤的重要机制之一。本研究对HPV16型E6蛋白诱导人原代角质形成细胞（primary human keratinocytes，PHK）形成多倍体以及对PHK有丝分裂纺锤体检查点的影响进行初步的探索。方法：取正常人包皮组织，分离表皮，常规方法培养PHK。脂质体介导法将pBabe-16E6质粒转染逆转录病毒包装细胞PA317。挑选G418抗性克隆，检测病毒滴度，将表达HPV16型E6蛋白的高滴度逆转录病毒感染PHK。通过逆转录PCR法和免疫印迹法证实HPV16E6成功感染PHK。PHK经Nocodazole处理后，利用流式细胞仪分析多倍体形成，于不同时间点收集细胞、固定，进行抗磷酸化组蛋白染色，利用流式细胞仪分析细胞有丝分裂指数差异。结果：HPV16型E6成功感染PHK，并呈功能性表达。HPV16型E6可诱导PHK形成多倍体，表达E6蛋白的PHK和对照细胞以相似的动力学进入和退出有丝分裂。结论：HPV16型E6对PHK有丝分裂过程中的纺锤体检查点无直接影响，鉴于有丝分裂后期检查点在细胞有丝分裂过程中较持久和严格的作用，本研究结果提示，对有丝分裂后期检查点的作用可能是HPV16型E6蛋白诱导宿主细胞多倍体形成的重要机制。为进一步探讨HPV相关肿瘤的分子发生机制奠定了一定基础。展开更多
文摘Using inducers to induce cells to produce inflammatory response is a common in vitro experimental method to study inflammation.However,there are relatively few inflammatory models developed for the cosmetic industry,and there are also great differences in the control of model induction,the selection of inflammatory indicators,and the concentration of inducers.Therefore,in this paper,by systematically studying the effects of Lipopolysaccharide(LPS)on the cell viability,the levels of IL-1α,IL-8 and ROS of human primary keratinocytes,a skin inflammation model based human primary keratinocyte was developed.The results showed that 0.01~100μg/mL LPS had no significant effect on the cell viability of human primary keratinocytes,while 100μg/mL LPS could simultaneously induce human primary keratinocytes to produce large amounts of IL-1αand IL-8,and 0.01μg/mL LPS could induce plentiful ROS.Therefore,a skin inflammation model for differential induction of different inflammatory indicators was established,and the sample OSM2021041301 was tested with this model,it was found that sample OSM2021041301 could significantly inhibit LPS-induced elevated IL-1αand IL-8 levels,the inhibitory effect on LPS-induced elevated ROS level was weak.The results indicated that OSM2021041301 has certain anti-inflammatory effect on inflammation caused by the increase of IL-1α,IL-8 and ROS induced by LPS and its analogues.
文摘目的：人乳头瘤病毒16型（human papillomavirus，HPV 16）与包括宫颈癌在内的多种肿瘤的发生明确相关。诱导宿主基因组不稳定性可能是HPV16致瘤的重要机制之一。本研究对HPV16型E6蛋白诱导人原代角质形成细胞（primary human keratinocytes，PHK）形成多倍体以及对PHK有丝分裂纺锤体检查点的影响进行初步的探索。方法：取正常人包皮组织，分离表皮，常规方法培养PHK。脂质体介导法将pBabe-16E6质粒转染逆转录病毒包装细胞PA317。挑选G418抗性克隆，检测病毒滴度，将表达HPV16型E6蛋白的高滴度逆转录病毒感染PHK。通过逆转录PCR法和免疫印迹法证实HPV16E6成功感染PHK。PHK经Nocodazole处理后，利用流式细胞仪分析多倍体形成，于不同时间点收集细胞、固定，进行抗磷酸化组蛋白染色，利用流式细胞仪分析细胞有丝分裂指数差异。结果：HPV16型E6成功感染PHK，并呈功能性表达。HPV16型E6可诱导PHK形成多倍体，表达E6蛋白的PHK和对照细胞以相似的动力学进入和退出有丝分裂。结论：HPV16型E6对PHK有丝分裂过程中的纺锤体检查点无直接影响，鉴于有丝分裂后期检查点在细胞有丝分裂过程中较持久和严格的作用，本研究结果提示，对有丝分裂后期检查点的作用可能是HPV16型E6蛋白诱导宿主细胞多倍体形成的重要机制。为进一步探讨HPV相关肿瘤的分子发生机制奠定了一定基础。
