H.pylori对人肝正常细胞L-02的作用及其分子机制  

Effect of Helicobacter pylori infection on growth of normal human liver cells(L-02)

作　　者：刘丽[1] 黄赞松[1,2] 黄衍强[3] 周喜汉[2] 

机构地区：[1]广西医科大学研究生学院,广西壮族自治区南宁市530021 [2]右江民族医学院消化疾病研究所附属医院消化内科,广西壮族自治区百色市533000 [3]右江民族医学院微生物免疫学教研室,广西壮族自治区百色市533000

出　　处：《世界华人消化杂志》2014年第7期956-962,共7页World Chinese Journal of Digestology

基　　金：广西自然科学基金资助项目;No.桂科自0542119;广西教育厅重点课题基金资助项目;No.桂教研重201202ZD078;广西医疗卫生重点科研课题基金资助项目;No.桂卫重200887~~

摘　　要：目的:研究幽门螺杆菌(Helicobacter pylori,H.pylori)对人肝正常细胞L-02(human normal liver cell line-02)的作用及转化生长因子β受体Ⅰ(transforming growth factor-beta typeⅠ,TβRⅠ)基因表达的影响.方法:体外培养L-02细胞、H.pylori;PCR法鉴定CagA+H.pylori和CagA-H.pylori,采用不同浓度的CagA+H.pylori和CagA-H.pylori作用于L-02细胞24 h,并设不加H.pylori的L-02细胞为阴性对照组;MTT法检测L-02细胞抑制率,通过Real-time PCR法检测各组TβRⅠ基因的表达.结果:MTT法结果显示,随着CagA+H.pylori、CagA-H.pylori浓度升高,对L-02细胞的抑制作用增强,与阴性对照组相比,差别均有统计学意义(P<0.05);在同一浓度,CagA+H.pylori对细胞的抑制作用较CagA-H.pylori明显,两组抑制率(%)分别为101 CFU/mL组10.960±0.231 vs 4.470±0.289;102 CFU/mL组25.310±0.398 vs 5.510±0.168;103 CFU/mL组33.130±0.312 vs 10.330±0.213;104 CFU/mL组54.570±0.245 vs 17.120±0.309;105 CFU/mL组79.450±0.402 vs 25.830±0.337;106 CFU/mL组90.210±0.271 vs 32.350±0.178,各组比较均t<0.05;Real-time PCR法检测中,随着CagA+H.pylori、CagA-H.pylori浓度升高,TβRⅠ表达逐渐升高,在同一浓度,CagA+H.pylori促TβRⅠ表达作用较CagA-H.pylori明显,两组TβRⅠ相对表达量分别为101 CFU/mL组1.65±0.101 vs 1.110±0.110;102 CFU/mL组2.770±0.198 vs 1.200±0.203;103 CFU/mL组4.590±0.112 vs 1.590±0.134;104 CFU/mL组5.470±0.145 vs 1.990±0.331;105 CFU/mL组7.450±0.102 vs 2.650±0.268;106 CFU/mL组8.570±0.221 vs 4.570±0.161,各组比较均t<0.05.结论:H.pylori对人肝正常细胞L-02细胞具有抑制作用并与浓度相关,菌液浓度越高,抑制细胞增殖作用明显,CagA+H.pylori的抑制作用比CagA-H.pylori强.H.pylori对L-02细胞作用后TβRⅠ基因表达增高并与浓度相关,CagA+H.pylori的影响比CagA-H.pylori大.H.pylori对L-02细胞抑制作用的可能机制是通过TβRⅠ基因表达改变干扰转化生长因子-β1/Smads信号通路传导而影响其生长.AIM： To investigate the effect of Helicobacter pylori（H. pylori） infection on the growth of humannormal liver cell line-02（L-02） and the possible mechanisms involved. METHODS： L-02 cells and H. pylori strains were cultured in vitro. CagA^＋ and CagA^- H. pylori strains were identified by PCR. After L-02 cells were incubated with different densities of CagA^＋ and CagA^- H. pylori strains for 24 h, MTT assay was used to detect the inhibition rate of L-02 cells, and the expression of transforming growth factor-beta typeⅠ（TβRⅠ） in L-02 cells was detected by real-time PCR. Cells without H. pylori infection were used as a negative control group. RESULTS： Both CagA^＋ and CagA^- H. pylori strains significantly inhibited L-02 cell growth in a dose-dependent manner compared with the control group（P〈0.05）. At the same concentration, CagA^＋ H. pylori strain had a more significant inhibitory effect on L-02 cell growth than CagA^＋ H. pylori strain（10^1 CFU/mL： 10.96 ± 0.231 vs 4.47 ± 0.289; 10^2 CFU/mL： 25.31 ± 0.398 vs 5.51 ± 0.168; 10^3 CFU/mL： 33.13 ± 0.312 vs 10.33 ± 0.213; 10^4 CFU/mL： 54.57 ± 0.245 vs 17.12 ± 0.309; 10^5 CFU/mL： 79.45 ± 0.402 vs 25.83 ± 0.337; 10^6 CFU/mL： 90.21 ± 0.271 vs 32.35 ± 0.178; t〈0.05 for all）. Both CagA^＋ and CagA^- H. pylori strains significantly increased the expression of TβRⅠ in a dose-dependent manner compared with the control group. At the same concentration, CagA^＋ H. pylori strain increased the expression of TβR Ⅰmore significantly than CagA^- H. pylori strain（10^1 CFU/mL： 1.65 ± 0.101 vs 1.11 ± 0.110; 10^2 CFU/mL： 2.77 ± 0.198 vs 1.20 ± 0.203; 10^3 CFU/ mL： 4.59 ± 0.112 vs 1.59 ± 0.134; 10^4 CFU/mL： 5.47 ± 0.145 vs 1.99 ± 0.331; 10^5 CFU/mL： 7.45 ± 0.102 vs 2.65 ± 0.268; 10^6 CFU/mL： 8.57 ± 0.221 vs 4.57 ± 0.161; t 〈0.05 for all）. CONCLUSION： H. pylori infection inhibits the growth of L-02 cells in a dose-dependent manner, and the inhibitory effect of CagA^＋ H. pylori strain

关 键 词：幽门螺杆菌 CAGA L-02细胞 转化生长因子β受体Ⅰ 

分 类 号：R346[医药卫生—基础医学]