HAX-1对MRL/lpr小鼠狼疮活动性影响的研究

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.

Effects of HS1-associated protein X-1 on the lupus activities:experiment、vith MRL/lpr lupus-like mice

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,iniected 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 iniection 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.MTF 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 MTY method was used to detect the proliferation rateof the spleen lymphocytes.Results The urine protein 1evel 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 glomemli 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 CHIVe of the spleen lymphocytes of Group C was higher than those of Groups A and B.The spleen lymphoeyte 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.