摘要
目的观察抗凋亡蛋白HAX-1对MRL／lpr狼疮鼠狼疮活动性的影响。方法重组腺病毒AdHAX-1经腹腔注射MRL／lpr小鼠，每周2次，连续注射4周。分别在注射前、注射2周后及4周后检测小鼠血清中抗核抗体（ANA）、循环免疫复合物（CIC）、抗双链DNA抗体（抗dsDNA抗体）、抗组蛋白抗体和干扰素γ（IFN-γ）水平，并检测注射前及注射后外周血的白细胞计数及尿蛋白水平；经RNAi使HAX-1表达下调后，观察脾脏淋巴细胞的增殖以及IFN-γ的分泌水平。结果HAX-1可使MRL／lpr小鼠的尿蛋白以及抗dsDNA抗体、ANA、抗组蛋白抗体、CIC明显升高，且产生IFN-γ水平升高，除抗组蛋白抗体及CIC外，其余指标与磷酸盐缓冲液对照组（PBS组）及对照腺病毒组（AdEGFP组）相比较，差异均有统计学意义（均P〈0．05）。重组腺病毒组（AdHax-1组）小鼠肾小球内免疫复合物沉积强度明显较强且阳性肾小球数目较多，与对照组比较差异均有统计学意义（均P〈0．05）。AdHax-1组肾小球的平均病变积分达1．50±0．34，与对照组（PBS组：0．67±0．14；AdEGFP组：0．81±0．26）比较差异均有统计学意义（均P〈0．01）。pGenesil-HAX-1可使淋巴细胞的增殖率明显降低（P〈0．05），IFN-γ分泌水平明显下降（P〈0．01）。结论HAX-1是调节系统性红斑狼疮淋巴细胞凋亡的一个重要因素，重组AdHAX-1可使MRL／lpr小鼠狼疮样症状加重，下调HAX-1表达可使病情减轻。
Objective To investigate the effects of HS1-associated protein X-1 （HAX-1） on the lupus activity of MRL/lpr lupus-like mice. Methods Fifteen MRL/lpr mice were divided into 3 equal groups： Group A, injected with phosphate-buffered saline, Group B, injected intraperitoneally with control virus AdEGFP and Group C, injected intraperitoneally with recombinant AdHAX-1 twice a week for 4 weeks. Peripheral blood samples were collected before the injection, and 2 and 4 weeks after the injection to be detected the white blood cell count, antinuclear antibody （ ANA）, anti-double strand-DNA antibodies, circulating immune complex （CIC） , anti-histone antibodies, and interferon （IFN） -γ. The level of urine protein was measured, too. Then the mice were killed, a kidney underwent direct immunofluorescence （DIF） to observe the deposition of Immune complexes, and the other kidney underwent periodic acid-Schiff （PAS） staining and pathological examination. MTT method was used to detect the proliferation of the lymphocytes in the spleen. Splenocytes were isolated from the other 15 MRL/lpr mice and then divided into 4 groups： Group, transfected with DMRIE-C without plasmid; Group E, as negative control group; Group F, transfected with blank plasmid pGenesil-1 ; and Group G, transfected with pGenesil-HAX-1. Forty-eight hours later MTT method was used to detect the proliferation rateof the spleen lymphocytes. Results The urine protein level of Group C was significantly higher than those of Groups A and B （ both P 〈0.01 ）. Four weeks later the levels of ANA, anti-double strand DNA antibodies, and IFN-γ were all significantly higher than those of Groups A and B （all P 〈0.01）. Hypercellularity and increased deposition of IgG in glomeruli were also observed in Group C. The score of glomeruli lesion of Group C （1.50 ± 0. 34） was significantly higher than those of Groups A （0.67±0.14） and Groups B （0.81±0.26） （both P〈0.01）. MTT method showed that the growth curve of the spleen lymphocytes of Group C was higher than those of Groups A and B. The spleen lymphocyte proliferation rate and the levels of IFN-γ of Group G was significantly lower than that of Group F （both P 〈0.05）. Conclusion One of the important factors in apoptosis regulation of SLE, HAX-1 may be involved in the pathogenesis of SLE, and the silence of HAX-1 may be beneficial for the improvement of SLE.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2008年第6期406-410,共5页
National Medical Journal of China
基金
国家自然科学基金资助项目（30671886）
