基于轉錄因子T-bet、GATA3的IRPS雙向免疫調節(jié)機理的研究
發(fā)布時間：2019-02-11 09:29
【摘要】：為了闡明板藍根多糖雙向免疫調節(jié)的內在機理,確定雙向免疫調節(jié)的關鍵因子,為中藥的雙向免疫調節(jié)作用提供理論支持,本試驗用不同劑量的環(huán)磷酰胺以不同的注射方式建立免疫抑制和免疫亢進大鼠模型,并以該大鼠模型為媒介在不同的時間點從轉錄因子、細胞因子以及機體免疫三方面來研究板藍根多糖對不同免疫狀態(tài)大鼠的影響,試驗分5部分,如下所述:試驗一不同劑量的IRPS灌胃大鼠,通過檢測淋巴細胞增殖活性,NK細胞活性以及血清中抗體含量確定了IRPS作用于大鼠最佳劑量為60mg/kg,為后續(xù)進一步的試驗研究做了準備。試驗二為了建立免疫抑制以及免疫亢進動物模型,本試驗用不同劑量的Cy在不同的時間以不同的方式作用于大鼠,并從免疫器官、免疫細胞、抗體和細胞因子等方面對所造模型進行評價,結果發(fā)現在腹腔注射OVA后6h腹腔一次性注射Cy125mg/kg或100mg/kg能構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,能夠構建良好的免疫亢進模型。試驗三劑量為60mg/kg的IRPS灌胃不同免疫狀態(tài)的大鼠,并在免疫的第6d和第12d,檢測大鼠T淋巴細胞和B淋巴細胞的增殖情況,NK細胞的活性以及抗體分泌量,結果表明在免疫第6d,IRPS對不同免疫狀態(tài)大鼠B淋巴細胞有雙向免疫調節(jié)作用,對免疫狀態(tài)正常大鼠和免疫抑制大鼠T淋巴細胞增殖活性影響不顯著;在免疫第12d,IRPS主要發(fā)揮免疫增強作用,能夠增強免疫正常狀態(tài)大鼠和免疫亢進大鼠機體的免疫功能,對于免疫抑制大鼠發(fā)揮雙向調節(jié)作用的同時也有一定的免疫增強作用。試驗四劑量為60mg/kg的IRPS灌胃不同免疫狀態(tài)的大鼠,在不同的時間點檢測不同免疫狀態(tài)大鼠Th1/Th2型細胞因子以及炎性因子的含量,結果表明IRPS能夠緩解免疫亢進組大鼠Th1/Th2型細胞因子的降低趨勢,對免疫抑制組大鼠Th1/Th2型細胞因子的升高趨勢也有一定的抑制作用,對不同免疫狀態(tài)大鼠炎性因子的分泌有雙向調節(jié)作用,說明IRPS能夠依據機體所處免疫狀態(tài)的不同對細胞因子表達發(fā)揮不同的調節(jié)作用。試驗五劑量為60mg/kg的IRPS灌胃不同免疫狀態(tài)的大鼠,在不同的時間點檢測不同免疫狀態(tài)大鼠轉錄因子T-bet和GATA3mRNA的表達,結果表明IRPS對免疫抑制大鼠T-bet/GATA3的比值有一個明顯的雙向調節(jié)作用,同時也能夠降低免疫狀態(tài)正常大鼠Tbet/GATA3的比值。上述試驗結果表明:IRPS能夠通過對細胞因子分泌和轉錄因子表達的影響進而對免疫亢進和免疫抑制大鼠發(fā)揮雙向免疫調節(jié)作用,對健康大鼠發(fā)揮一定的免疫保護作用。同時我們的研究結果也為進一步研究IRPS發(fā)揮其免疫功能的靶點的確定提供了參考。
[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.
【學位授予單位】：河南農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S853.7
本文編號：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.
【學位授予單位】：河南農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S853.7
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