CAJ | 학술논문

目的探讨腹腔注射姜黄素对小鼠移植心脏存活时间的影响及其机制.方法选择C57BL/6和Babl/c小鼠各30只,分别作为心脏移植的供、受者,进行小鼠腹部心脏移植.随机将受者分为对照组、小剂量组和大剂量组,在术前1 d至术后7 d每天分别给受者腹腔注射生理盐水、5 μg/μl和10 μg/μl的姜黄素溶液,注射量均为20μl/g.术后观察各组移植心脏存活时间;术后第7天,每组分别取3只受者的移植心脏行病理学检查,观察排斥反应情况.并采用酶联免疫吸附试验(ELISA)法检测受者血清白细胞介素2(IL-2)、IL-4、IL-10及γ干扰素(IFN-γ)的水平,采用混合淋巴细胞反应检测受者T淋巴细胞对供者抗原的反应性,分别采用免疫印迹法和ELISA法检测受者T淋巴细胞核因子κB(NF-κB)p65核转位和NF-κB p65 DNA的结合活性.结果对照组、小剂量组和大剂量组移植心存活时间分别为(8.0±3.3)d、(17.0±2.4)d和(23.0±2.2)d,三组间的差异均有统计学意义(P＜0.01).术后第7天,对照组、小剂量组和大剂量组移植心排斥反应分级分别为Ⅱ～Ⅲ、Ⅰ～Ⅱ和0～Ⅰ级.对照组、小剂量组和大剂量组血清IL-2和IFN-γ的水平依次降低,而血清IL-4和IL-10的水平依次增高,三组间各细胞因子的差异均有统计学意义(P＜0.01).小剂昔组和大剂量组的T淋巴细胞经供者T淋巴细胞刺激后,其增殖能力减弱,大剂量组尤其明显.小剂量组和大剂量组的T淋巴细胞NF-κB p65核转位减少,NF-κB 065 DNA的结合活性降低.大剂量组的该效应明显强于小剂量组(P＜0.01).结论姜黄素能显著延长小鼠移植心脏存活时间,抑制心脏移植排斥反应,其机制可能是姜黄素能够抑制TH1型细胞和促进TH2型细胞因子的表达,使受者对供者抗原处于免疫低反应状态.
목적 탐토복강주사강황소대소서이식심장존활시간적영향급기궤제.방법 선택C57BL/6화Babl/c소서각30지,분별작위심장이식적공、수자,진행소서복부심장이식.수궤장수자분위대조조、소제량조화대제량조,재술전1 d지술후7 d매천분별급수자복강주사생리염수、5 μg/μl화10 μg/μl적강황소용액,주사량균위20μl/g.술후관찰각조이식심장존활시간;술후제7천,매조분별취3지수자적이식심장행병이학검사,관찰배척반응정황.병채용매련면역흡부시험(ELISA)법검측수자혈청백세포개소2(IL-2)、IL-4、IL-10급γ간우소(IFN-γ)적수평,채용혼합림파세포반응검측수자T림파세포대공자항원적반응성,분별채용면역인적법화ELISA법검측수자T림파세포핵인자κB(NF-κB)p65핵전위화NF-κB p65 DNA적결합활성.결과 대조조、소제량조화대제량조이식심존활시간분별위(8.0±3.3)d、(17.0±2.4)d화(23.0±2.2)d,삼조간적차이균유통계학의의(P＜0.01).술후제7천,대조조、소제량조화대제량조이식심배척반응분급분별위Ⅱ～Ⅲ、Ⅰ～Ⅱ화0～Ⅰ급.대조조、소제량조화대제량조혈청IL-2화IFN-γ적수평의차강저,이혈청IL-4화IL-10적수평의차증고,삼조간각세포인자적차이균유통계학의의(P＜0.01).소제석조화대제량조적T림파세포경공자T림파세포자격후,기증식능력감약,대제량조우기명현.소제량조화대제량조적T림파세포NF-κB p65핵전위감소,NF-κB 065 DNA적결합활성강저.대제량조적해효응명현강우소제량조(P＜0.01).결론 강황소능현저연장소서이식심장존활시간,억제심장이식배척반응,기궤제가능시강황소능구억제TH1형세포화촉진TH2형세포인자적표체,사수자대공자항원처우면역저반응상태.
Objective To approach the effects of injecting intraperitoneally with curcumin on the survival time of mouse cardiac allografts and the mechanism.Methods Heterotopic cardiac transplantations of mice were done by using 30 C57 mice as donors,and 30 Babl/C mice as recipients.The recipients were classified randomly into three groups:control group,low-dose curcumin group,and high-dose curcumin group.The recipients of the corresponding groups were injected intraperitoneally with sodium chloride,5 μg/μl curcumin,and 10 μg/μl curcumin respectively.The injecting volume of all mice was 20 μl/g.Survival time of transplanted hearts was recorded after the operations.The grafts of 3 mice in each group were harvested for histopathological examination 7 days after the operations,ELISA was used to determine the changes of IL-2,IL-4.IL-10,IFN-γ in peripheral blood of the receptors,the mixed lympholeukocyte reaction was done to inspect responsibility of T lymphocytes of receptors to donor antigen,Western-blot was used to examine nuclear translocation of NF-κB p65 in T cells,and ELISA to analyze DNA binding activity of NF-κB p65 in T cells.Results The average survival time of mouse heart allografts in control group,low-dose curcumin group and high-dose curcumin group was(8.0 ± 3.3),(17.0 ± 2.4),and(23.0 ± 2.2),respectively(P＜0.01).On the 7th day after operations,the grade of rejecting reactions in control group,low-dose curcumin group,and high-dose curcumin group was Ⅱ-Ⅲ,Ⅰ-Ⅱ and 0-Ⅰ,correspondingly.IL-2,and IFN-γ in peripheral blood of receptors in control group,low-dose curcumin group,and high-dose curcumin group were reduced in turn,but IL-4 and IL-10 were increased in turn (P＜0.01).As compared with control group,the proliferation ability of T cells in low-dose curcumin group and high-dose curcumin group was attenuated,especially in high-dose curcumin group,when stimulated with splenocytes of donators.Translocation of NF-κB p65 and NF-κB DNA binding activity of T cells in low-dose curcumin group and high-dose curcumin group were decreased.Conclusion Curcurmin can obviously prolong the survival time of allografts and inhibit the rejecting reactions of mouse cardiac allografts possibly by inhibiting cytokines in Th1 cells and promoting cytokines in Th2 cells,making the receptor immunosuppression against donor antigen.