Anti-RANTES聯(lián)合環(huán)孢素A誘導(dǎo)小鼠二次心臟移植長期免疫耐受
發(fā)布時間：2018-12-10 08:38
【摘要】：目的探討Anti-RANTES聯(lián)合環(huán)孢素A在小鼠心臟二次移植急性排斥反應(yīng)中的作用。方法構(gòu)建小鼠心臟二次移植模型:初次移植,以Balb/c小鼠為供鼠,C57BL/6小鼠為受鼠,進行腹部心臟移植;二次移植,以同種Balb/c小鼠為供鼠,初次移植后存活的C57BL/6小鼠為受鼠,進行二次頸部心臟移植。然后經(jīng)不同步驟處理,分為四組進行實驗:對照組:初次腹部心臟移植存活小鼠,于初次腹部心臟移植2w后行二次頸部心臟移植,同時予腹腔注射與其他組同劑量的生理鹽水處理;實驗組A,初次腹部心臟移植存活小鼠,于初次腹部心臟移植2w后行心臟二次頸部移植,同時予腹腔注射與其他組同劑量的環(huán)孢素A處理(n=6);實驗組B,初次腹部心臟移植存活小鼠,于初次腹部心臟移植2w后行心臟二次頸部移植,同時予腹腔注射與其他組同劑量的Anti-RANTES處理(n=6);實驗組C,初次腹部心臟移植存活小鼠,于初次腹部心臟移植2w后行心臟二次頸部移植,同時予腹腔混合注射與其他組同劑量的環(huán)孢素A和Anti-RANTES處理(n=6)。觀察四組二次移植心臟存活時間;HE染色組織病理學(xué)改變觀察各組異位心臟急性排斥反應(yīng)的程度;Q-PCR檢測二次心臟移植物中RANTES、IFN-γ、IL-2、IL-10和TGF-β基因的相對表達量;ELISA檢測RANTES、IFN-γ、IL-2、IL-10和TGF-β在受鼠血清中的濃度。結(jié)果在Anti-RANTES聯(lián)合環(huán)孢素A實驗組,移植心臟的存活時間可達8.58±0.24天,而在對照組、Anti-RANTES組和應(yīng)用環(huán)孢素A組,移植心臟的存活時間分別為(3.1±0.43)、(5.2±0.26)和(5.58±0.20)天(P0.01);心臟移植物HE染色可見,在Anti-RANTES聯(lián)合環(huán)孢素A實驗組,其炎癥細胞的浸潤較對照組、單用Anti-RANTES實驗組A、單環(huán)孢素A實驗組B明顯減少(P0.01);Q-PCR的結(jié)果提示,移植心臟內(nèi)RANTES、IL-2、INF-γ的m RNA表達在Anti-RANTES聯(lián)合環(huán)孢素A實驗組較其他組明顯減少,而移植心臟內(nèi)IL-10、TGF-βm RNA的表達在Anti-RANTES聯(lián)合環(huán)孢素A實驗組較其他組明顯增加(P0.01);ELISA的結(jié)果提示,移植心臟內(nèi)RANTES、IL-2、INF-γ的血清濃度在Anti-RANTES聯(lián)合環(huán)孢素A實驗組較其他組明顯減少,而移植心臟內(nèi)IL-10、TGF-β血清濃度在Anti-RANTES聯(lián)合環(huán)孢素A實驗組較其他組明顯增加(P0.01)。結(jié)論CC族趨化因子配體5(CCL5)RANTES,在小鼠二次心臟移植免疫排斥的誘發(fā)和發(fā)展過程中,起著至關(guān)重要的作用。單用可以Anti-RANTES或者環(huán)孢素A都可以有效抑制二次心臟移植的急性排斥反應(yīng),減輕炎癥細胞向移植物聚集、減輕移植物中炎癥細胞因子RANTES、IL-2、INF-γ的分泌而增加抗炎細胞因子IL-10、TGF-β的分泌,從而延長二次移植物的存活時間。但聯(lián)合應(yīng)用Anti-RANTES和環(huán)孢素A,以上抗急性免疫排斥效果更加明顯,因而可以誘導(dǎo)小鼠對二次心臟移植較長期耐受。
[Abstract]:Objective to investigate the role of Anti-RANTES combined with cyclosporine A in acute rejection of second heart transplantation in mice. Methods the second heart transplantation model of mice was established. The abdominal heart transplantation was carried out with Balb/c mice as donor and C57BL/6 mice as recipient mice for the first time. The second transplantation was carried out with the allogeneic Balb/c mice as donor mice and the C57BL/6 mice survived after the first transplantation as recipient mice. Then, after different steps, they were divided into four groups: the control group: the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, At the same time, intraperitoneal injection of the same dose of normal saline treatment as other groups; Experimental group A, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of cyclosporine A treatment (nong6); The experimental group B, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of Anti-RANTES as other groups (nong6); Experimental group C, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of cyclosporine A and Anti-RANTES treatment (NN6). The survival time of the cardiac allograft in the four groups and the degree of acute rejection of ectopic heart in each group were observed by HE staining histopathological changes. Q-PCR was used to detect the relative expression of RANTES,IFN- 緯, IL-2,IL-10 and TGF- 尾 in secondary heart grafts, and ELISA was used to detect the concentrations of RANTES,IFN- 緯, IL-2,IL-10 and TGF- 尾 in the serum of recipient mice. Results the survival time of transplanted heart was 8.58 鹵0.24 days in Anti-RANTES combined with cyclosporine A group, while in control group, Anti-RANTES group and cyclosporine A group, the survival time was (3.1 鹵0.43). (5.2 鹵0.26) and (5.58 鹵0.20) days (P0.01); HE staining of cardiac grafts showed that the infiltration of inflammatory cells in Anti-RANTES combined with cyclosporine A group was significantly lower than that in control group. The number of inflammatory cells in Anti-RANTES group and single cyclosporine A group B was significantly lower than that in control group (P0.01). The results of Q-PCR showed that the expression of m RNA of RANTES,IL-2,INF- 緯 in the transplanted heart was significantly lower in the experimental group of Anti-RANTES combined with cyclosporine A than in the other groups, but IL-10, in the transplanted heart. The expression of TGF- 尾 m RNA in Anti-RANTES combined with cyclosporine A group was significantly higher than that in other groups (P0.01). The results of ELISA showed that the serum concentration of RANTES,IL-2,INF- 緯 in the transplanted heart was significantly lower in the experimental group of Anti-RANTES combined with cyclosporine A than that in the other groups, but IL-10, in the transplanted heart. The serum concentration of TGF- 尾 in Anti-RANTES combined with cyclosporine A group was significantly higher than that in other groups (P0.01). Conclusion CC chemokine ligand 5 (CCL5) RANTES, plays an important role in the induction and development of immune rejection after secondary heart transplantation in mice. Anti-RANTES or cyclosporine A alone can effectively inhibit the acute rejection of secondary heart transplantation, reduce the accumulation of inflammatory cells to the grafts, and alleviate the inflammatory cytokines RANTES,IL-2, in the grafts. The secretion of INF- 緯 increased the secretion of anti inflammatory cytokine IL-10,TGF- 尾, thus prolonging the survival time of the secondary graft. But the combination of Anti-RANTES and cyclosporine A had more obvious effect on acute immune rejection, so it could induce long-term tolerance to secondary heart transplantation in mice.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R654.2
本文編號：2370296
[Abstract]:Objective to investigate the role of Anti-RANTES combined with cyclosporine A in acute rejection of second heart transplantation in mice. Methods the second heart transplantation model of mice was established. The abdominal heart transplantation was carried out with Balb/c mice as donor and C57BL/6 mice as recipient mice for the first time. The second transplantation was carried out with the allogeneic Balb/c mice as donor mice and the C57BL/6 mice survived after the first transplantation as recipient mice. Then, after different steps, they were divided into four groups: the control group: the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, At the same time, intraperitoneal injection of the same dose of normal saline treatment as other groups; Experimental group A, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of cyclosporine A treatment (nong6); The experimental group B, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of Anti-RANTES as other groups (nong6); Experimental group C, the first abdominal heart transplantation survival mice, after the first abdominal heart transplantation 2 weeks after the second cervical heart transplantation, at the same time intraperitoneal injection of the same dose of cyclosporine A and Anti-RANTES treatment (NN6). The survival time of the cardiac allograft in the four groups and the degree of acute rejection of ectopic heart in each group were observed by HE staining histopathological changes. Q-PCR was used to detect the relative expression of RANTES,IFN- 緯, IL-2,IL-10 and TGF- 尾 in secondary heart grafts, and ELISA was used to detect the concentrations of RANTES,IFN- 緯, IL-2,IL-10 and TGF- 尾 in the serum of recipient mice. Results the survival time of transplanted heart was 8.58 鹵0.24 days in Anti-RANTES combined with cyclosporine A group, while in control group, Anti-RANTES group and cyclosporine A group, the survival time was (3.1 鹵0.43). (5.2 鹵0.26) and (5.58 鹵0.20) days (P0.01); HE staining of cardiac grafts showed that the infiltration of inflammatory cells in Anti-RANTES combined with cyclosporine A group was significantly lower than that in control group. The number of inflammatory cells in Anti-RANTES group and single cyclosporine A group B was significantly lower than that in control group (P0.01). The results of Q-PCR showed that the expression of m RNA of RANTES,IL-2,INF- 緯 in the transplanted heart was significantly lower in the experimental group of Anti-RANTES combined with cyclosporine A than in the other groups, but IL-10, in the transplanted heart. The expression of TGF- 尾 m RNA in Anti-RANTES combined with cyclosporine A group was significantly higher than that in other groups (P0.01). The results of ELISA showed that the serum concentration of RANTES,IL-2,INF- 緯 in the transplanted heart was significantly lower in the experimental group of Anti-RANTES combined with cyclosporine A than that in the other groups, but IL-10, in the transplanted heart. The serum concentration of TGF- 尾 in Anti-RANTES combined with cyclosporine A group was significantly higher than that in other groups (P0.01). Conclusion CC chemokine ligand 5 (CCL5) RANTES, plays an important role in the induction and development of immune rejection after secondary heart transplantation in mice. Anti-RANTES or cyclosporine A alone can effectively inhibit the acute rejection of secondary heart transplantation, reduce the accumulation of inflammatory cells to the grafts, and alleviate the inflammatory cytokines RANTES,IL-2, in the grafts. The secretion of INF- 緯 increased the secretion of anti inflammatory cytokine IL-10,TGF- 尾, thus prolonging the survival time of the secondary graft. But the combination of Anti-RANTES and cyclosporine A had more obvious effect on acute immune rejection, so it could induce long-term tolerance to secondary heart transplantation in mice.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R654.2
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1 羅增榮;Anti-RANTES聯(lián)合環(huán)孢素A誘導(dǎo)小鼠二次心臟移植長期免疫耐受[D];福建醫(yī)科大學(xué);2015年
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