【摘要】：為了闡明板藍(lán)根多糖雙向免疫調(diào)節(jié)的內(nèi)在機(jī)理,確定雙向免疫調(diào)節(jié)的關(guān)鍵因子,為中藥的雙向免疫調(diào)節(jié)作用提供理論支持,本試驗(yàn)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(jìn)大鼠模型,并以該大鼠模型為媒介在不同的時(shí)間點(diǎn)從轉(zhuǎn)錄因子、細(xì)胞因子以及機(jī)體免疫三方面來研究板藍(lán)根多糖對(duì)不同免疫狀態(tài)大鼠的影響,試驗(yàn)分5部分,如下所述:試驗(yàn)一不同劑量的IRPS灌胃大鼠,通過檢測(cè)淋巴細(xì)胞增殖活性,NK細(xì)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(jìn)一步的試驗(yàn)研究做了準(zhǔn)備。試驗(yàn)二為了建立免疫抑制以及免疫亢進(jìn)動(dòng)物模型,本試驗(yàn)用不同劑量的Cy在不同的時(shí)間以不同的方式作用于大鼠,并從免疫器官、免疫細(xì)胞、抗體和細(xì)胞因子等方面對(duì)所造模型進(jìn)行評(píng)價(jià),結(jié)果發(fā)現(xiàn)在腹腔注射OVA后6h腹腔一次性注射Cy125mg/kg或100mg/kg能構(gòu)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(gòu)建良好的免疫亢進(jìn)模型。試驗(yàn)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(cè)大鼠T淋巴細(xì)胞和B淋巴細(xì)胞的增殖情況,NK細(xì)胞的活性以及抗體分泌量,結(jié)果表明在免疫第6d,IRPS對(duì)不同免疫狀態(tài)大鼠B淋巴細(xì)胞有雙向免疫調(diào)節(jié)作用,對(duì)免疫狀態(tài)正常大鼠和免疫抑制大鼠T淋巴細(xì)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(qiáng)作用,能夠增強(qiáng)免疫正常狀態(tài)大鼠和免疫亢進(jìn)大鼠機(jī)體的免疫功能,對(duì)于免疫抑制大鼠發(fā)揮雙向調(diào)節(jié)作用的同時(shí)也有一定的免疫增強(qiáng)作用。試驗(yàn)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(shí)間點(diǎn)檢測(cè)不同免疫狀態(tài)大鼠Th1/Th2型細(xì)胞因子以及炎性因子的含量,結(jié)果表明IRPS能夠緩解免疫亢進(jìn)組大鼠Th1/Th2型細(xì)胞因子的降低趨勢(shì),對(duì)免疫抑制組大鼠Th1/Th2型細(xì)胞因子的升高趨勢(shì)也有一定的抑制作用,對(duì)不同免疫狀態(tài)大鼠炎性因子的分泌有雙向調(diào)節(jié)作用,說明IRPS能夠依據(jù)機(jī)體所處免疫狀態(tài)的不同對(duì)細(xì)胞因子表達(dá)發(fā)揮不同的調(diào)節(jié)作用。試驗(yàn)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(shí)間點(diǎn)檢測(cè)不同免疫狀態(tài)大鼠轉(zhuǎn)錄因子T-bet和GATA3mRNA的表達(dá),結(jié)果表明IRPS對(duì)免疫抑制大鼠T-bet/GATA3的比值有一個(gè)明顯的雙向調(diào)節(jié)作用,同時(shí)也能夠降低免疫狀態(tài)正常大鼠Tbet/GATA3的比值。上述試驗(yàn)結(jié)果表明:IRPS能夠通過對(duì)細(xì)胞因子分泌和轉(zhuǎn)錄因子表達(dá)的影響進(jìn)而對(duì)免疫亢進(jìn)和免疫抑制大鼠發(fā)揮雙向免疫調(diào)節(jié)作用,對(duì)健康大鼠發(fā)揮一定的免疫保護(hù)作用。同時(shí)我們的研究結(jié)果也為進(jìn)一步研究IRPS發(fā)揮其免疫功能的靶點(diǎn)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(xué)位授予單位】：河南農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S853.7

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