目的检测柯萨奇B3病毒(coxsackievirus group B type3,CVB3)感染体外培养的星形胶质细胞后集聚蛋白(agrin)的表达变化及其意义。方法采用免疫细胞化学的方法鉴定培养的星形胶质细胞,在显微镜下观察CVB3感染后的细胞形态,并用RT-PCR方法...目的检测柯萨奇B3病毒(coxsackievirus group B type3,CVB3)感染体外培养的星形胶质细胞后集聚蛋白(agrin)的表达变化及其意义。方法采用免疫细胞化学的方法鉴定培养的星形胶质细胞,在显微镜下观察CVB3感染后的细胞形态,并用RT-PCR方法扩增CVB3RNA的正负链以确证感染成功,用RT-PCR检测CVB3感染细胞后agrin的动态表达变化。用脂质体将agrin反义质粒pcDNA3-As-AG转染到星形胶质细胞,并用RT-PCR检测转染的细胞中agrin的表达。3H-TdR掺入法测定CVB3不同感染时间点的星形胶质细胞裂解液和经agrin反义质粒pcDNA-As-AG转染细胞后的细胞裂解液对T细胞增殖的影响。结果(1)CVB3感染星形胶质细胞后agrin的表达水平下调。(2)病毒感染后随着agrin表达量的降低,星形胶质细胞裂解液对T细胞的活化增殖作用也降低。未转染和空质粒转染的细胞裂解液对T细胞的活化增殖有促进作用,而用反义核酸技术下调星形胶质细胞agrin表达后,星形胶质细胞裂解液对T细胞增殖的促进作用也随之下降,进一步证实agrin表达水平的下调可影响T细胞的活化和增殖。结论CVB3感染星形胶质细胞后agrin表达水平下调,降低对T细胞活化增殖的能力,这可能是CVB3病毒性脑炎中,病毒逃逸机体免疫清除的一种机制。展开更多
Common additives in plastics such as bisphenol A (BPA) or phthalates like di-(2-ethylhexyl) phthalate (DEHP) are environmental estrogens that have been shown to be endocrine disruptors in some experimental animal mode...Common additives in plastics such as bisphenol A (BPA) or phthalates like di-(2-ethylhexyl) phthalate (DEHP) are environmental estrogens that have been shown to be endocrine disruptors in some experimental animal models. This project used the C2C12 cell culture model to examine how exposure to BPA or DEHP affects two aspects of skeletal muscle development, the fusion of myoblasts into myotubes and agrin-induced clustering of acetylcholine receptors (AChRs). During myotube formation AChRs cluster spontaneously. Treatment with motor neuron derived agrin increases the frequency of AChR clusters through an agrin signaling pathway that also clusters other postsynaptic components of the neuromuscular synapse. For this project C2C12 cells were exposed to BPA or DEHP while myoblasts fused into myotubes. After exposure to 10 μM BPA or 100 μM DEHP the frequency of agrin-induced AChR clusters decreased. In addition, myotube formation decreased as a higher percentage of nuclei remained in myoblasts. Furthermore, BPA or DEHP reduced the amount of the myogenic regulatory factor myogenin. This suggests that BPA and DEHP decrease AChR clustering by reducing myogenin. Moreover, plastic additives like BPA and DEHP may pose a risk for skeletal muscle development in humans.展开更多
Objective:To study the effects and potential control mechanisms of the agrin signaling pathway on the repair of distal sciatic nerve damage.Methods:50 Sprague-Dawleyrats were randomly allocated into the control group(...Objective:To study the effects and potential control mechanisms of the agrin signaling pathway on the repair of distal sciatic nerve damage.Methods:50 Sprague-Dawleyrats were randomly allocated into the control group(SCI group,n?25)and the treatment group(Treat group,n?25).In both groups,the sciatic nerve damage model was created.Following surgery,the SCI group and the Treat group received intragastric administrations of normal saline and Jiawei Buyang Huanwu Decoction,respectively.The dose for both groups was 2mL/(kg$d)for 12 consecutive weeks.At 4,8,and 12 weeks post-surgery,the wet-to-weight ratio of the gastrocnemius muscle in the two groups was assessed;and hematoxylin-eosin staining and Masson staining were used to detect pathological alterations in the sciatic nerve and collagen.The expression of agrin,Dag1,Dmd,and Lamb2 proteins in the agrin pathway,as well as ERK,MEK,and their corresponding phosphorylated proteins,p-ERK and p-MEK,were all detected by Western blot in the gastrocnemius muscle.Results:The wet weight of the gastrocnemius muscle in the Treat group increased considerably(p<0.05)after 4,8,and 12 weeks following the operation compared to the SCI group,and it continued to rise over time.The expression level of Dag1,Dmd,and Lamb2 protein in agrin pathway,as well as p-ERK and p-MEK protein were all significantly lower than they were in the SCI group(p<0.05).Additionally,Jiawei Buyang Huanwu Decoction greatly reduced the amount of nerve scars at the sciatic nerve injury and significantly healed sciatic neuropathy according to the results of pathological staining.Conclusion:Jiawei Buyang Huanwu Decoction can inhibit the expression of agrin,Dag1,Dmd,and Lamb2,as well as p-ERK and p-MEK.It can effectively promote the distal repair of sciatic nerve injury probably through argin pathway.展开更多
转移、侵袭和免疫逃避是肿瘤的重要生物学特性,聚集蛋白Agrin结合低密度脂蛋白受体相关蛋白4(low density density lipoprotein receptor related protein 4,Lrp4),参与肿瘤的侵袭与迁移.此外,Agrin还促进血管形成,并通过α-肌萎缩蛋白...转移、侵袭和免疫逃避是肿瘤的重要生物学特性,聚集蛋白Agrin结合低密度脂蛋白受体相关蛋白4(low density density lipoprotein receptor related protein 4,Lrp4),参与肿瘤的侵袭与迁移.此外,Agrin还促进血管形成,并通过α-肌萎缩蛋白聚糖(α-dystroglycan)促进T细胞免疫应答,稳定免疫突触,在肿瘤的免疫微环境中扮演着重要的角色.深入研究恶性肿瘤以及免疫细胞中的Agrin,探索其作用机制,可能为肿瘤的免疫治疗提供新方向.现就Agrin的研究现状,特别是其在恶性肿瘤与免疫作用的相关方面研究进行总结与分析.展开更多
文摘目的检测柯萨奇B3病毒(coxsackievirus group B type3,CVB3)感染体外培养的星形胶质细胞后集聚蛋白(agrin)的表达变化及其意义。方法采用免疫细胞化学的方法鉴定培养的星形胶质细胞,在显微镜下观察CVB3感染后的细胞形态,并用RT-PCR方法扩增CVB3RNA的正负链以确证感染成功,用RT-PCR检测CVB3感染细胞后agrin的动态表达变化。用脂质体将agrin反义质粒pcDNA3-As-AG转染到星形胶质细胞,并用RT-PCR检测转染的细胞中agrin的表达。3H-TdR掺入法测定CVB3不同感染时间点的星形胶质细胞裂解液和经agrin反义质粒pcDNA-As-AG转染细胞后的细胞裂解液对T细胞增殖的影响。结果(1)CVB3感染星形胶质细胞后agrin的表达水平下调。(2)病毒感染后随着agrin表达量的降低,星形胶质细胞裂解液对T细胞的活化增殖作用也降低。未转染和空质粒转染的细胞裂解液对T细胞的活化增殖有促进作用,而用反义核酸技术下调星形胶质细胞agrin表达后,星形胶质细胞裂解液对T细胞增殖的促进作用也随之下降,进一步证实agrin表达水平的下调可影响T细胞的活化和增殖。结论CVB3感染星形胶质细胞后agrin表达水平下调,降低对T细胞活化增殖的能力,这可能是CVB3病毒性脑炎中,病毒逃逸机体免疫清除的一种机制。
文摘Common additives in plastics such as bisphenol A (BPA) or phthalates like di-(2-ethylhexyl) phthalate (DEHP) are environmental estrogens that have been shown to be endocrine disruptors in some experimental animal models. This project used the C2C12 cell culture model to examine how exposure to BPA or DEHP affects two aspects of skeletal muscle development, the fusion of myoblasts into myotubes and agrin-induced clustering of acetylcholine receptors (AChRs). During myotube formation AChRs cluster spontaneously. Treatment with motor neuron derived agrin increases the frequency of AChR clusters through an agrin signaling pathway that also clusters other postsynaptic components of the neuromuscular synapse. For this project C2C12 cells were exposed to BPA or DEHP while myoblasts fused into myotubes. After exposure to 10 μM BPA or 100 μM DEHP the frequency of agrin-induced AChR clusters decreased. In addition, myotube formation decreased as a higher percentage of nuclei remained in myoblasts. Furthermore, BPA or DEHP reduced the amount of the myogenic regulatory factor myogenin. This suggests that BPA and DEHP decrease AChR clustering by reducing myogenin. Moreover, plastic additives like BPA and DEHP may pose a risk for skeletal muscle development in humans.
基金Funding Science and Technology Development Project of Traditional Chinese Medicine in Shandong Province(Grant No.2019-0500).
文摘Objective:To study the effects and potential control mechanisms of the agrin signaling pathway on the repair of distal sciatic nerve damage.Methods:50 Sprague-Dawleyrats were randomly allocated into the control group(SCI group,n?25)and the treatment group(Treat group,n?25).In both groups,the sciatic nerve damage model was created.Following surgery,the SCI group and the Treat group received intragastric administrations of normal saline and Jiawei Buyang Huanwu Decoction,respectively.The dose for both groups was 2mL/(kg$d)for 12 consecutive weeks.At 4,8,and 12 weeks post-surgery,the wet-to-weight ratio of the gastrocnemius muscle in the two groups was assessed;and hematoxylin-eosin staining and Masson staining were used to detect pathological alterations in the sciatic nerve and collagen.The expression of agrin,Dag1,Dmd,and Lamb2 proteins in the agrin pathway,as well as ERK,MEK,and their corresponding phosphorylated proteins,p-ERK and p-MEK,were all detected by Western blot in the gastrocnemius muscle.Results:The wet weight of the gastrocnemius muscle in the Treat group increased considerably(p<0.05)after 4,8,and 12 weeks following the operation compared to the SCI group,and it continued to rise over time.The expression level of Dag1,Dmd,and Lamb2 protein in agrin pathway,as well as p-ERK and p-MEK protein were all significantly lower than they were in the SCI group(p<0.05).Additionally,Jiawei Buyang Huanwu Decoction greatly reduced the amount of nerve scars at the sciatic nerve injury and significantly healed sciatic neuropathy according to the results of pathological staining.Conclusion:Jiawei Buyang Huanwu Decoction can inhibit the expression of agrin,Dag1,Dmd,and Lamb2,as well as p-ERK and p-MEK.It can effectively promote the distal repair of sciatic nerve injury probably through argin pathway.
文摘转移、侵袭和免疫逃避是肿瘤的重要生物学特性,聚集蛋白Agrin结合低密度脂蛋白受体相关蛋白4(low density density lipoprotein receptor related protein 4,Lrp4),参与肿瘤的侵袭与迁移.此外,Agrin还促进血管形成,并通过α-肌萎缩蛋白聚糖(α-dystroglycan)促进T细胞免疫应答,稳定免疫突触,在肿瘤的免疫微环境中扮演着重要的角色.深入研究恶性肿瘤以及免疫细胞中的Agrin,探索其作用机制,可能为肿瘤的免疫治疗提供新方向.现就Agrin的研究现状,特别是其在恶性肿瘤与免疫作用的相关方面研究进行总结与分析.
