中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2010年
10期
740-744
,共5页
肝硬化,胆汁性%受体,雌激素%多态性,单核苷酸%T淋巴细胞亚群%细胞因子类
肝硬化,膽汁性%受體,雌激素%多態性,單覈苷痠%T淋巴細胞亞群%細胞因子類
간경화,담즙성%수체,자격소%다태성,단핵감산%T림파세포아군%세포인자류
Liver cirrhosis,biliary%Receptors,estrogen%Polymorphism,single nucleotide%T-lymphocyte subsets%Cytokines
目的 探讨雌激素受体(ER)α基因多态性对女性原发性胆汁淤积性肝硬化(PBC)患者T淋巴细胞亚群细胞因子漂移的影响.方法 以未经治疗的60例女性PBC患者为研究组,性别、年龄匹配的健康体检者52名为对照组进行研究.采用聚合酶链反应-限制性片段长度多态性方法检测ER α基因1号内含子PvuⅡ和Xba Ⅰ酶切位点的多态性.流式细胞仪定量检测外周血CD4+、CD8+、CD4+CD25+、CD4+CD28-T淋巴细胞.RT-PCR方法检测外周血单个核细胞肿瘤坏死因子(TNF)α、白细胞介素(IL)-2、干扰素(IFN)v、IL-4、IL-6、IL-10 mRNA的表达.基因型和等位基因频率采用直接基因计数法计算,并进行Hardy-Weinberg遗传平衡检验与Chi-square x2检验;两组间计量资料采用t或t'检验.结果 女性PBC患者的ER α基因PvuⅡ酶切基因亚型中Pp的阳性率明显高于正常对照组,pp基因亚型的阳性率则明显低于对照组(x2=7.2880,P＜0.05).Xba Ⅰ基因型以及各等位基因频率在女性PBC组与对照组的差异无统计学意义.PBC患者的ppxx基因型的阳性率低于正常对照组,但两组间差异无统计学意义(x2=6.5382,P＞0.05).相对于对照组的33.81%±3.87%,PBC患者T淋巴细胞中CD4+T淋巴细胞比例增高为45.31%±5.26%(t=7.86,P＜0.01),CD8+T淋巴细胞比例由对照组的31.83%±1.73%降低为27.78%±1.43%(t=8.24,P＜0.01).与对照组相比,PBC患者CD4+CD25+T淋巴细胞比例明显降低; CD4+CD28-T淋巴细胞比例则明显升高.PBC患者外周血单个核细胞TNF α、IL-2和IFN γ mRNA表达水平分别为0.59±0.19、0.71±0.29、0.67±0.21,明显高于对照组的0.22±0.13、0.31±0.14、0.27±0.13(t值分别为6.93,5.07,7.01,P值均＜0.01);IL-6 mRNA表达水平为0.45±0.21,高于对照组的0.34±0.16(t=1.84,P＜0.05); IL-4和IL-10 mRNA表达水平组间差异无统计学意义.结论 PvuⅡ基因的Pp基因型是女性PBC的遗传易感基因型,p等位基因是易感基因.PBC患者外`周血以Th1细胞亚群及其细胞因子占优势.ER α基因多态性可作为遗传易感背景影响PBC患者体内T淋巴细胞亚群的偏移及其相关细胞因子的表达.
目的 探討雌激素受體(ER)α基因多態性對女性原髮性膽汁淤積性肝硬化(PBC)患者T淋巴細胞亞群細胞因子漂移的影響.方法 以未經治療的60例女性PBC患者為研究組,性彆、年齡匹配的健康體檢者52名為對照組進行研究.採用聚閤酶鏈反應-限製性片段長度多態性方法檢測ER α基因1號內含子PvuⅡ和Xba Ⅰ酶切位點的多態性.流式細胞儀定量檢測外週血CD4+、CD8+、CD4+CD25+、CD4+CD28-T淋巴細胞.RT-PCR方法檢測外週血單箇覈細胞腫瘤壞死因子(TNF)α、白細胞介素(IL)-2、榦擾素(IFN)v、IL-4、IL-6、IL-10 mRNA的錶達.基因型和等位基因頻率採用直接基因計數法計算,併進行Hardy-Weinberg遺傳平衡檢驗與Chi-square x2檢驗;兩組間計量資料採用t或t'檢驗.結果 女性PBC患者的ER α基因PvuⅡ酶切基因亞型中Pp的暘性率明顯高于正常對照組,pp基因亞型的暘性率則明顯低于對照組(x2=7.2880,P＜0.05).Xba Ⅰ基因型以及各等位基因頻率在女性PBC組與對照組的差異無統計學意義.PBC患者的ppxx基因型的暘性率低于正常對照組,但兩組間差異無統計學意義(x2=6.5382,P＞0.05).相對于對照組的33.81%±3.87%,PBC患者T淋巴細胞中CD4+T淋巴細胞比例增高為45.31%±5.26%(t=7.86,P＜0.01),CD8+T淋巴細胞比例由對照組的31.83%±1.73%降低為27.78%±1.43%(t=8.24,P＜0.01).與對照組相比,PBC患者CD4+CD25+T淋巴細胞比例明顯降低; CD4+CD28-T淋巴細胞比例則明顯升高.PBC患者外週血單箇覈細胞TNF α、IL-2和IFN γ mRNA錶達水平分彆為0.59±0.19、0.71±0.29、0.67±0.21,明顯高于對照組的0.22±0.13、0.31±0.14、0.27±0.13(t值分彆為6.93,5.07,7.01,P值均＜0.01);IL-6 mRNA錶達水平為0.45±0.21,高于對照組的0.34±0.16(t=1.84,P＜0.05); IL-4和IL-10 mRNA錶達水平組間差異無統計學意義.結論 PvuⅡ基因的Pp基因型是女性PBC的遺傳易感基因型,p等位基因是易感基因.PBC患者外`週血以Th1細胞亞群及其細胞因子佔優勢.ER α基因多態性可作為遺傳易感揹景影響PBC患者體內T淋巴細胞亞群的偏移及其相關細胞因子的錶達.
목적 탐토자격소수체(ER)α기인다태성대녀성원발성담즙어적성간경화(PBC)환자T림파세포아군세포인자표이적영향.방법 이미경치료적60례녀성PBC환자위연구조,성별、년령필배적건강체검자52명위대조조진행연구.채용취합매련반응-한제성편단장도다태성방법검측ER α기인1호내함자PvuⅡ화Xba Ⅰ매절위점적다태성.류식세포의정량검측외주혈CD4+、CD8+、CD4+CD25+、CD4+CD28-T림파세포.RT-PCR방법검측외주혈단개핵세포종류배사인자(TNF)α、백세포개소(IL)-2、간우소(IFN)v、IL-4、IL-6、IL-10 mRNA적표체.기인형화등위기인빈솔채용직접기인계수법계산,병진행Hardy-Weinberg유전평형검험여Chi-square x2검험;량조간계량자료채용t혹t'검험.결과 녀성PBC환자적ER α기인PvuⅡ매절기인아형중Pp적양성솔명현고우정상대조조,pp기인아형적양성솔칙명현저우대조조(x2=7.2880,P＜0.05).Xba Ⅰ기인형이급각등위기인빈솔재녀성PBC조여대조조적차이무통계학의의.PBC환자적ppxx기인형적양성솔저우정상대조조,단량조간차이무통계학의의(x2=6.5382,P＞0.05).상대우대조조적33.81%±3.87%,PBC환자T림파세포중CD4+T림파세포비례증고위45.31%±5.26%(t=7.86,P＜0.01),CD8+T림파세포비례유대조조적31.83%±1.73%강저위27.78%±1.43%(t=8.24,P＜0.01).여대조조상비,PBC환자CD4+CD25+T림파세포비례명현강저; CD4+CD28-T림파세포비례칙명현승고.PBC환자외주혈단개핵세포TNF α、IL-2화IFN γ mRNA표체수평분별위0.59±0.19、0.71±0.29、0.67±0.21,명현고우대조조적0.22±0.13、0.31±0.14、0.27±0.13(t치분별위6.93,5.07,7.01,P치균＜0.01);IL-6 mRNA표체수평위0.45±0.21,고우대조조적0.34±0.16(t=1.84,P＜0.05); IL-4화IL-10 mRNA표체수평조간차이무통계학의의.결론 PvuⅡ기인적Pp기인형시녀성PBC적유전역감기인형,p등위기인시역감기인.PBC환자외`주혈이Th1세포아군급기세포인자점우세.ER α기인다태성가작위유전역감배경영향PBC환자체내T림파세포아군적편이급기상관세포인자적표체.
Objective To study the effects of estrogen receptor (ER) alpha gene polymorphism on the migration of T lymphocyte subsets and related cytokines in the female patients with primary biliary cirrhosis (PBC). Methods This study was conducted with sixty female PBC patients without treatment as the study group and fifty-two healthy people wtih sex and age met the requirements of the study as the control group.The polymorphism of restriction enzyme cutting site of Xba Ⅰ and Pvu Ⅱ in intron 1 of ER α gene was detected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). CD4+, CD8+,CD4+CD25+ and CD4+CD28 T lymphocytes in peripheral blood were quantitatively detected by flow cytometry.RT-PCR method was used to detect the expression of TNF α, IL-2, IFN γ, IL-4, IL-6 and IL-10 in peripheral mononuclear cells. Results The positive rate of Pp in ER α gene Pvu Ⅱ enzyme gene subtypes of female PBC patients was significantly greater than that of the control group, and the positive rate of pp gene subtype was significantly smaller than that of the control group ( x2 = 7.2880, P = 0.0261). The difference of Xba Ⅰ genotype and allele frequency between the female PBC patients group and the control group was not of statistical significance ( x 2 = 6.5382, P = 0.5833). The proportion of CD4+ T in T lymphocytes of PBC patients was increased to 45.31% ± 5.26%, compared with 33.81% ± 3.87% in the control group; and the proportion of CD8+T lymphocytes was decreased to 27.78% ± 1.43 % from 31.83% ± 1.73% in the control group. In comparison with the control group, the proportion of CD4+CD25+ T lymphocytes decreased significantly, while that of CD4+CD28- T lymphocytes rose significantly. The expression levels of TNF α,IL-2 and IFN γ mRNA were 0.59 ± 0.19, 0.71 ± 0.29 and 0.67 ± 0.21 respectively, which were significantly higher than that in the control group (0.22 ± 0.13, 0.31 ± 0.14, 0.27 ± 0.13) (t = 6.93, 5.07, 7.01,P ＜ 0.01); the expression level of IL-6 mRNA was increased to 0.45 ± 0.21 from 0.34 ± 0.16 in the control group (t = 1.84, P ＜ 0.05); and the difference of the expression levels of IL-4 and IL-10 mRNA between two groups was not of statistical significance. Conclusion Pp of gene Pvu Ⅱ was a genetically susceptible genotype in female PBC patients, and the allele p was a susceptible gene. Th1 cell subsets and related cytokines were dominant in peripheral blood of PBC patients. As a background of genetic susceptibility, ER α gene polymorphism could affect the shift of T lymphocyte subsets and the expression of the related cytokines in PBC patients.
