中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2011年
11期
833-837
,共5页
周耀勇%邹勇%陈韬%王洪武%韩梅芳%皮斌%严伟明%习东%黄加权%罗小平%宁琴
週耀勇%鄒勇%陳韜%王洪武%韓梅芳%皮斌%嚴偉明%習東%黃加權%囉小平%寧琴
주요용%추용%진도%왕홍무%한매방%피빈%엄위명%습동%황가권%라소평%저금
肝炎,病毒性,动物%T淋巴细胞%NK细胞%KCTD9
肝炎,病毒性,動物%T淋巴細胞%NK細胞%KCTD9
간염,병독성,동물%T림파세포%NK세포%KCTD9
Hepatitis,viral,animal%T-lymphocyte%NK cells%Potassium channel tetramerisation domain containing 9
目的 初步探讨KCTD9在暴发性肝炎模型小鼠中的作用机制.方法 78只BALB/cJ小鼠(其中雄性6只)随机分为暴发性肝炎对照组和暴发性肝炎模型组,每组39只(每组雄性3只);75只C3H/HeJ雌性小鼠分为慢性肝炎对照组(36只)和慢性肝炎模型组(39只).模型组每只小鼠腹腔注射MHV-3(100 PFU).暴发性肝炎对照组和模型组48 h试验点以1只雄性小鼠供取睾丸,2只雌性小鼠供取肝脾等组织标本和分离肝细胞,10只雌性小鼠为1个单位取肝组织供分离肝脾淋巴细胞；慢性肝炎对照组和模型组48 h时以1只小鼠供取肝脾组织,1只小鼠供分离肝细胞,10只为1个单位供分离肝脾淋巴细胞,重复3次,剩下3只小鼠在建模15d时处死,仅供取肝脾组织.磁珠分选肝脾单个核细胞,采用免疫组织化学、实时定量PCR技术检测KCTD9在暴发性肝炎、慢性肝炎模型动物中的表达差异,各组间比较用t检验或方差分析.结果 与对照组比较,KCTD9 mRNA在暴发性肝炎模型组小鼠肝CD8+T细胞、CD4+T细胞、自然杀伤细胞(NK)细胞中分别上调8.8倍、59.4倍、577.1倍,t值分别为-5.393、-3.706和-4.714,P值均＜0.05,差异有统计学意义;在脾CD4+T细胞、CD8+T细胞中,KCTD9 mRNA表达水平分别下调了43％和69％,t值分别为2.906、10.98,P值均＜ 0.05,差异有统计学意义.与对照组比较,在慢性肝炎模型组小鼠肝NK细胞和CD4+T细胞中KCTD9 mRNA分别下调71％、51％,t值分别为3.827、5.746,P值均＜0.05,差异有统计学意义.KCTD9蛋白在暴发性肝炎模型组小鼠肝细胞中呈弱阳性表达,坏死灶周围及组织中炎症浸润细胞中呈强阳性表达；脾中阳性细胞仍散在分布,与对照组比较,脾白髓中细胞数量减少达71％.结论 KCTD9在MHV-3诱导的暴发性肝炎模型小鼠肝脏高表达,可能参与疾病发生和发展过程.
目的 初步探討KCTD9在暴髮性肝炎模型小鼠中的作用機製.方法 78隻BALB/cJ小鼠(其中雄性6隻)隨機分為暴髮性肝炎對照組和暴髮性肝炎模型組,每組39隻(每組雄性3隻);75隻C3H/HeJ雌性小鼠分為慢性肝炎對照組(36隻)和慢性肝炎模型組(39隻).模型組每隻小鼠腹腔註射MHV-3(100 PFU).暴髮性肝炎對照組和模型組48 h試驗點以1隻雄性小鼠供取睪汍,2隻雌性小鼠供取肝脾等組織標本和分離肝細胞,10隻雌性小鼠為1箇單位取肝組織供分離肝脾淋巴細胞；慢性肝炎對照組和模型組48 h時以1隻小鼠供取肝脾組織,1隻小鼠供分離肝細胞,10隻為1箇單位供分離肝脾淋巴細胞,重複3次,剩下3隻小鼠在建模15d時處死,僅供取肝脾組織.磁珠分選肝脾單箇覈細胞,採用免疫組織化學、實時定量PCR技術檢測KCTD9在暴髮性肝炎、慢性肝炎模型動物中的錶達差異,各組間比較用t檢驗或方差分析.結果 與對照組比較,KCTD9 mRNA在暴髮性肝炎模型組小鼠肝CD8+T細胞、CD4+T細胞、自然殺傷細胞(NK)細胞中分彆上調8.8倍、59.4倍、577.1倍,t值分彆為-5.393、-3.706和-4.714,P值均＜0.05,差異有統計學意義;在脾CD4+T細胞、CD8+T細胞中,KCTD9 mRNA錶達水平分彆下調瞭43％和69％,t值分彆為2.906、10.98,P值均＜ 0.05,差異有統計學意義.與對照組比較,在慢性肝炎模型組小鼠肝NK細胞和CD4+T細胞中KCTD9 mRNA分彆下調71％、51％,t值分彆為3.827、5.746,P值均＜0.05,差異有統計學意義.KCTD9蛋白在暴髮性肝炎模型組小鼠肝細胞中呈弱暘性錶達,壞死竈週圍及組織中炎癥浸潤細胞中呈彊暘性錶達；脾中暘性細胞仍散在分佈,與對照組比較,脾白髓中細胞數量減少達71％.結論 KCTD9在MHV-3誘導的暴髮性肝炎模型小鼠肝髒高錶達,可能參與疾病髮生和髮展過程.
목적 초보탐토KCTD9재폭발성간염모형소서중적작용궤제.방법 78지BALB/cJ소서(기중웅성6지)수궤분위폭발성간염대조조화폭발성간염모형조,매조39지(매조웅성3지);75지C3H/HeJ자성소서분위만성간염대조조(36지)화만성간염모형조(39지).모형조매지소서복강주사MHV-3(100 PFU).폭발성간염대조조화모형조48 h시험점이1지웅성소서공취고환,2지자성소서공취간비등조직표본화분리간세포,10지자성소서위1개단위취간조직공분리간비림파세포；만성간염대조조화모형조48 h시이1지소서공취간비조직,1지소서공분리간세포,10지위1개단위공분리간비림파세포,중복3차,잉하3지소서재건모15d시처사,부공취간비조직.자주분선간비단개핵세포,채용면역조직화학、실시정량PCR기술검측KCTD9재폭발성간염、만성간염모형동물중적표체차이,각조간비교용t검험혹방차분석.결과 여대조조비교,KCTD9 mRNA재폭발성간염모형조소서간CD8+T세포、CD4+T세포、자연살상세포(NK)세포중분별상조8.8배、59.4배、577.1배,t치분별위-5.393、-3.706화-4.714,P치균＜0.05,차이유통계학의의;재비CD4+T세포、CD8+T세포중,KCTD9 mRNA표체수평분별하조료43％화69％,t치분별위2.906、10.98,P치균＜ 0.05,차이유통계학의의.여대조조비교,재만성간염모형조소서간NK세포화CD4+T세포중KCTD9 mRNA분별하조71％、51％,t치분별위3.827、5.746,P치균＜0.05,차이유통계학의의.KCTD9단백재폭발성간염모형조소서간세포중정약양성표체,배사조주위급조직중염증침윤세포중정강양성표체；비중양성세포잉산재분포,여대조조비교,비백수중세포수량감소체71％.결론 KCTD9재MHV-3유도적폭발성간염모형소서간장고표체,가능삼여질병발생화발전과정.
Objective To explore the mechanisms of a novel potassium channel gene named KCTD9 (potassium channel tetramerisation domain containing 9) in model of fulminant viral hepatitis induced by murine hepatitis virus 3 (MHV-3).Methods 78 BALB/cJ mice(6 male) were randomly and equally assigned to two groups,model group of fulminant viral hepatitis induced by MHV3 and its control.75 C3HAHeJ female mice were done into two groups,39 for model group of chronic hepatitis induced by MHV3,36 for control.Various samples including spleen,liver and lyrnphocytes from mice of two model groups and the controls were examined for KCTD9 expression by real time quantitative PCR and Immunohistochemistry.Independent-samples T test or one-way ANOVA were carried out in different groups.Results Increased expressions of KCTD9 mRNA was observed in livers of both model mice of fulminant viral hepatitis and chronic hepatitis.Compared with the control mice,the expressions of KCTD9 mRNA were up-regulated by 577.1-,8.8-,59.4- and 10.8-fold in hepatic NK cells,CD4+ T cells,CD8+ T cells and splenic NK cells respectively in model mice of fulminant viral hepatitis 48hr post MHV-3 infection,whereas down-regulation by 43％ and 69％ in splenic CD4 + T cells and CD8+ T cells were found respectively.In contrast,in model mice of chronic viral hepatitis the expressions of KCTD9 mRNA were down-regulated by71％ and 51％ in hepatic CD4+ T cells and NK cells,respectively.The expression of KCTD9 protein was mainly evidenced in infiltrative monouclear cells of liver as shown by immunohistochemistry.Basal expression was also investigated and showed constitutive expression of KCTD9 in brain,thymus and other organs in BALB/cJ mice.Conclusion A novel potassium channel gene KCTD9 was highly expressed in hepatic NK cells and T cells of fulminant hepatitis mice induced by MHV-3.
