基于转录因子T-bet、GATA3的IRPS双向免疫调节机理的研究
发布时间：2019-02-11 09:29
【摘要】：为了阐明板蓝根多糖双向免疫调节的内在机理,确定双向免疫调节的关键因子,为中药的双向免疫调节作用提供理论支持,本试验用不同剂量的环磷酰胺以不同的注射方式建立免疫抑制和免疫亢进大鼠模型,并以该大鼠模型为媒介在不同的时间点从转录因子、细胞因子以及机体免疫三方面来研究板蓝根多糖对不同免疫状态大鼠的影响,试验分5部分,如下所述:试验一不同剂量的IRPS灌胃大鼠,通过检测淋巴细胞增殖活性,NK细胞活性以及血清中抗体含量确定了IRPS作用于大鼠最佳剂量为60mg/kg,为后续进一步的试验研究做了准备。试验二为了建立免疫抑制以及免疫亢进动物模型,本试验用不同剂量的Cy在不同的时间以不同的方式作用于大鼠,并从免疫器官、免疫细胞、抗体和细胞因子等方面对所造模型进行评价,结果发现在腹腔注射OVA后6h腹腔一次性注射Cy125mg/kg或100mg/kg能构建良好的免疫抑制模型,在免疫OVA前3d腹腔一次性注射Cy20mg/kg,能够构建良好的免疫亢进模型。试验三剂量为60mg/kg的IRPS灌胃不同免疫状态的大鼠,并在免疫的第6d和第12d,检测大鼠T淋巴细胞和B淋巴细胞的增殖情况,NK细胞的活性以及抗体分泌量,结果表明在免疫第6d,IRPS对不同免疫状态大鼠B淋巴细胞有双向免疫调节作用,对免疫状态正常大鼠和免疫抑制大鼠T淋巴细胞增殖活性影响不显著;在免疫第12d,IRPS主要发挥免疫增强作用,能够增强免疫正常状态大鼠和免疫亢进大鼠机体的免疫功能,对于免疫抑制大鼠发挥双向调节作用的同时也有一定的免疫增强作用。试验四剂量为60mg/kg的IRPS灌胃不同免疫状态的大鼠,在不同的时间点检测不同免疫状态大鼠Th1/Th2型细胞因子以及炎性因子的含量,结果表明IRPS能够缓解免疫亢进组大鼠Th1/Th2型细胞因子的降低趋势,对免疫抑制组大鼠Th1/Th2型细胞因子的升高趋势也有一定的抑制作用,对不同免疫状态大鼠炎性因子的分泌有双向调节作用,说明IRPS能够依据机体所处免疫状态的不同对细胞因子表达发挥不同的调节作用。试验五剂量为60mg/kg的IRPS灌胃不同免疫状态的大鼠,在不同的时间点检测不同免疫状态大鼠转录因子T-bet和GATA3mRNA的表达,结果表明IRPS对免疫抑制大鼠T-bet/GATA3的比值有一个明显的双向调节作用,同时也能够降低免疫状态正常大鼠Tbet/GATA3的比值。上述试验结果表明:IRPS能够通过对细胞因子分泌和转录因子表达的影响进而对免疫亢进和免疫抑制大鼠发挥双向免疫调节作用,对健康大鼠发挥一定的免疫保护作用。同时我们的研究结果也为进一步研究IRPS发挥其免疫功能的靶点的确定提供了参考。
[Abstract]:In order to elucidate the internal mechanism of bidirectional immunomodulation of Radix Isatidis polysaccharides, determine the key factors of bidirectional immune regulation, and provide theoretical support for the bidirectional immunomodulation of traditional Chinese medicine. In this study, different doses of cyclophosphamide were used to establish immunosuppressive and immune hyperactive rat models with different injection methods, and the rat model was used as a medium from transcription factors at different time points. The effects of Isatis lanceolata polysaccharides on different immune states of rats were studied in three aspects of cytokines and body immunity. The experiment was divided into five parts. The experiment was described as follows: experimental rats were given different doses of IRPS, and the proliferation activity of lymphocytes was measured. The activity of NK cells and the content of antibody in serum determined that the optimal dose of IRPS on rats was 60 mg / kg, which prepared for further experimental study. In experiment 2, in order to establish immunosuppressive and immune hyperactive animal models, different doses of Cy were used in different time and in different ways to act on rats, and from immune organs, immune cells, The antibody and cytokines were used to evaluate the model. The results showed that a good immunosuppressive model could be established by intraperitoneal injection of Cy125mg/kg or 100mg/kg 6 hours after intraperitoneal injection of OVA, and Cy20mg/kg, was injected intraperitoneally 3 days before immunization with OVA. It can construct a good model of immune hyperactivity. Three doses of IRPS with 60mg/kg were administered to rats with different immune states. The proliferation of T and B lymphocytes, the activity of NK cells and the amount of antibody secreted were measured on the 6th and 12th day after immunization. The results showed that IRPS had a bidirectional immunomodulatory effect on B lymphocytes in rats with different immune states on the 6th day after immunization, but had no significant effect on the proliferation activity of T lymphocytes in normal and immunosuppressed rats. IRPS can enhance immune function of normal and hyperimmunized rats on the 12th day, and it can also play a bidirectional regulatory role in immunosuppressive rats at the same time. Four doses of IRPS with 60mg/kg were administered to rats with different immune states. The levels of Th1/Th2 type cytokines and inflammatory cytokines were measured at different time points in rats. The results showed that IRPS could attenuate the decreasing trend of Th1/Th2 type cytokines in the hyperimmunized rats and inhibit the increasing trend of the Th1/Th2 type cytokines in the immunosuppressive group. It showed that IRPS could regulate cytokine expression according to the immune state of the body. The expression of transcription factor (T-bet) and GATA3mRNA in rats with different immune states were detected at different time points by intragastric administration of IRPS with 60mg/kg at different time points. The results showed that IRPS had a bidirectional regulatory effect on the ratio of T-bet/GATA3 in immunosuppressive rats and decreased the ratio of Tbet/GATA3 in normal immunized rats. The results showed that IRPS could exert bidirectional immunomodulatory effect on hyperimmune and immunosuppressive rats by influencing cytokine secretion and transcription factor expression, and had a certain immune protective effect on healthy rats. At the same time, our results also provide a reference for further study on the target of IRPS to play its immune function.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S853.7
本文编号：2419603
[Abstract]:In order to elucidate the internal mechanism of bidirectional immunomodulation of Radix Isatidis polysaccharides, determine the key factors of bidirectional immune regulation, and provide theoretical support for the bidirectional immunomodulation of traditional Chinese medicine. In this study, different doses of cyclophosphamide were used to establish immunosuppressive and immune hyperactive rat models with different injection methods, and the rat model was used as a medium from transcription factors at different time points. The effects of Isatis lanceolata polysaccharides on different immune states of rats were studied in three aspects of cytokines and body immunity. The experiment was divided into five parts. The experiment was described as follows: experimental rats were given different doses of IRPS, and the proliferation activity of lymphocytes was measured. The activity of NK cells and the content of antibody in serum determined that the optimal dose of IRPS on rats was 60 mg / kg, which prepared for further experimental study. In experiment 2, in order to establish immunosuppressive and immune hyperactive animal models, different doses of Cy were used in different time and in different ways to act on rats, and from immune organs, immune cells, The antibody and cytokines were used to evaluate the model. The results showed that a good immunosuppressive model could be established by intraperitoneal injection of Cy125mg/kg or 100mg/kg 6 hours after intraperitoneal injection of OVA, and Cy20mg/kg, was injected intraperitoneally 3 days before immunization with OVA. It can construct a good model of immune hyperactivity. Three doses of IRPS with 60mg/kg were administered to rats with different immune states. The proliferation of T and B lymphocytes, the activity of NK cells and the amount of antibody secreted were measured on the 6th and 12th day after immunization. The results showed that IRPS had a bidirectional immunomodulatory effect on B lymphocytes in rats with different immune states on the 6th day after immunization, but had no significant effect on the proliferation activity of T lymphocytes in normal and immunosuppressed rats. IRPS can enhance immune function of normal and hyperimmunized rats on the 12th day, and it can also play a bidirectional regulatory role in immunosuppressive rats at the same time. Four doses of IRPS with 60mg/kg were administered to rats with different immune states. The levels of Th1/Th2 type cytokines and inflammatory cytokines were measured at different time points in rats. The results showed that IRPS could attenuate the decreasing trend of Th1/Th2 type cytokines in the hyperimmunized rats and inhibit the increasing trend of the Th1/Th2 type cytokines in the immunosuppressive group. It showed that IRPS could regulate cytokine expression according to the immune state of the body. The expression of transcription factor (T-bet) and GATA3mRNA in rats with different immune states were detected at different time points by intragastric administration of IRPS with 60mg/kg at different time points. The results showed that IRPS had a bidirectional regulatory effect on the ratio of T-bet/GATA3 in immunosuppressive rats and decreased the ratio of Tbet/GATA3 in normal immunized rats. The results showed that IRPS could exert bidirectional immunomodulatory effect on hyperimmune and immunosuppressive rats by influencing cytokine secretion and transcription factor expression, and had a certain immune protective effect on healthy rats. At the same time, our results also provide a reference for further study on the target of IRPS to play its immune function.
【学位授予单位】：河南农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S853.7
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