中华消化杂志
中華消化雜誌
중화소화잡지
Chinese Journal of Digestion
2015年
3期
183-187
,共5页
许梦雀%曹海龙%王姗%曹晓沧%鄢芳%王邦茂
許夢雀%曹海龍%王姍%曹曉滄%鄢芳%王邦茂
허몽작%조해룡%왕산%조효창%언방%왕방무
脱氧胆酸%肠道菌群%癌变%Apcmin/+小鼠
脫氧膽痠%腸道菌群%癌變%Apcmin/+小鼠
탈양담산%장도균군%암변%Apcmin/+소서
Deoxycholic acid%Intestinal microbiota%Carcinogenesis%Apcmin/+ mice
目的 观察并分析次级胆汁酸诱导A pcmin/+小鼠肠道腺瘤癌变及其对肠道菌群的影响.方法 将20只4周龄Apcmin/+小鼠均分为A pcmin/+对照组、A pcmin/+脱氧胆酸组,将20只4周龄野生型C57BL/6J小鼠均分为野生型对照组、野生型脱氧胆酸组.对照组常规饮水,脱氧胆酸组饮用水中含0.2％脱氧胆酸.收集给药0周及12周A pcmin/+小鼠粪便,焦磷酸测序法分析肠道菌群变化.12周后处死,观察各组小鼠肠道腺瘤数目、大小及分布.采用HE染色评价腺瘤病理类型,采用免疫组织化学染色法检测增殖细胞核抗原(PCNA),采用脱氧核苷酸末端转移酶介导的dUTP缺口末端标记法(TUNEL)检测细胞凋亡.两组间计量资料的比较采用独立样本t检验.结果 所有野生型小鼠肠道均未出现肿瘤.Apcmin/+脱氧胆酸组小鼠肠道腺瘤总数较A pcmin/+对照组明显增加(57.00±3.07比21.50±4.69,t=20.03,P＜0.01),以最大径为1～2 mm的腺瘤增加最为显著(30.62±7.73比7.75±4.59,t=8.04,P＜0.05),腺瘤癌变率较Apcmin/+对照组也明显上升.Apcmin+脱氧胆酸组小鼠肠道肿瘤PCNA阳性细胞百分比较Apcmin/+对照组显著增加[(90.17±2.14)％比(41.97±4.26)％,t=31.97,P＜0.01)],肿瘤细胞凋亡百分比显著减少[(1.40±1.12)％比(7.50±0.65)％,t=14.90,P＜0.01].Apcmin/+脱氧胆酸组小鼠肠道菌群多样性明显降低,在菌门水平上厚壁菌门与拟杆菌门比值(0.586 7±0.148 4)较其对照组(0.387 3±0.013 6)明显升高(t=2.36,P＜0.05);在菌属及菌种水平上Apcmin/+脱氧胆酸组致病菌数量增多,益生菌数量明显降低.结论 脱氧胆酸能诱导Apcmin/+小鼠肠道菌群失衡,并可通过增加肠道肿瘤细胞增殖,抑制细胞凋亡促使肠道腺瘤发展成为肠癌.
目的 觀察併分析次級膽汁痠誘導A pcmin/+小鼠腸道腺瘤癌變及其對腸道菌群的影響.方法 將20隻4週齡Apcmin/+小鼠均分為A pcmin/+對照組、A pcmin/+脫氧膽痠組,將20隻4週齡野生型C57BL/6J小鼠均分為野生型對照組、野生型脫氧膽痠組.對照組常規飲水,脫氧膽痠組飲用水中含0.2％脫氧膽痠.收集給藥0週及12週A pcmin/+小鼠糞便,焦燐痠測序法分析腸道菌群變化.12週後處死,觀察各組小鼠腸道腺瘤數目、大小及分佈.採用HE染色評價腺瘤病理類型,採用免疫組織化學染色法檢測增殖細胞覈抗原(PCNA),採用脫氧覈苷痠末耑轉移酶介導的dUTP缺口末耑標記法(TUNEL)檢測細胞凋亡.兩組間計量資料的比較採用獨立樣本t檢驗.結果 所有野生型小鼠腸道均未齣現腫瘤.Apcmin/+脫氧膽痠組小鼠腸道腺瘤總數較A pcmin/+對照組明顯增加(57.00±3.07比21.50±4.69,t=20.03,P＜0.01),以最大徑為1～2 mm的腺瘤增加最為顯著(30.62±7.73比7.75±4.59,t=8.04,P＜0.05),腺瘤癌變率較Apcmin/+對照組也明顯上升.Apcmin+脫氧膽痠組小鼠腸道腫瘤PCNA暘性細胞百分比較Apcmin/+對照組顯著增加[(90.17±2.14)％比(41.97±4.26)％,t=31.97,P＜0.01)],腫瘤細胞凋亡百分比顯著減少[(1.40±1.12)％比(7.50±0.65)％,t=14.90,P＜0.01].Apcmin/+脫氧膽痠組小鼠腸道菌群多樣性明顯降低,在菌門水平上厚壁菌門與擬桿菌門比值(0.586 7±0.148 4)較其對照組(0.387 3±0.013 6)明顯升高(t=2.36,P＜0.05);在菌屬及菌種水平上Apcmin/+脫氧膽痠組緻病菌數量增多,益生菌數量明顯降低.結論 脫氧膽痠能誘導Apcmin/+小鼠腸道菌群失衡,併可通過增加腸道腫瘤細胞增殖,抑製細胞凋亡促使腸道腺瘤髮展成為腸癌.
목적 관찰병분석차급담즙산유도A pcmin/+소서장도선류암변급기대장도균군적영향.방법 장20지4주령Apcmin/+소서균분위A pcmin/+대조조、A pcmin/+탈양담산조,장20지4주령야생형C57BL/6J소서균분위야생형대조조、야생형탈양담산조.대조조상규음수,탈양담산조음용수중함0.2％탈양담산.수집급약0주급12주A pcmin/+소서분편,초린산측서법분석장도균군변화.12주후처사,관찰각조소서장도선류수목、대소급분포.채용HE염색평개선류병리류형,채용면역조직화학염색법검측증식세포핵항원(PCNA),채용탈양핵감산말단전이매개도적dUTP결구말단표기법(TUNEL)검측세포조망.량조간계량자료적비교채용독립양본t검험.결과 소유야생형소서장도균미출현종류.Apcmin/+탈양담산조소서장도선류총수교A pcmin/+대조조명현증가(57.00±3.07비21.50±4.69,t=20.03,P＜0.01),이최대경위1～2 mm적선류증가최위현저(30.62±7.73비7.75±4.59,t=8.04,P＜0.05),선류암변솔교Apcmin/+대조조야명현상승.Apcmin+탈양담산조소서장도종류PCNA양성세포백분비교Apcmin/+대조조현저증가[(90.17±2.14)％비(41.97±4.26)％,t=31.97,P＜0.01)],종류세포조망백분비현저감소[(1.40±1.12)％비(7.50±0.65)％,t=14.90,P＜0.01].Apcmin/+탈양담산조소서장도균군다양성명현강저,재균문수평상후벽균문여의간균문비치(0.586 7±0.148 4)교기대조조(0.387 3±0.013 6)명현승고(t=2.36,P＜0.05);재균속급균충수평상Apcmin/+탈양담산조치병균수량증다,익생균수량명현강저.결론 탈양담산능유도Apcmin/+소서장도균군실형,병가통과증가장도종류세포증식,억제세포조망촉사장도선류발전성위장암.
Objective To investigate secondary bile acid induced canceration process of intestinal adenoma and effects on intestinal microflora in Apcmin/+ mice.Methods Forty four-week-old mice (20 Apcmin/+mice and 20 wild-type C57BL/6J mice) were divided into four groups:wild-type control group (regular drinking water),wild-type deoxycholic acid (DOC) group (with 0.2 ％ DOC in drinking water),Apcmin/+ control group and Apcmin/+ DOC group.Fecal pellets of Apcmin/+ mice were collected at 0 week and 12 week after administration.The changes of intestinal microflora were analyzed by pyrosequencing.All mice were sacrificed after 12 weeks.The number,size and location of intestinal adenoma were observed.The pathological type of adenoma was evaluated after hematcxylin-eosin (HE) staining.Proliferating cell nuclear antigen (PCNA) was detected by immunohistochemistry.Cell apoptosis was determined by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling technique (TUNEL).Independent t test was used for the quantitative data comparison between two groups.Results No intestinal tumors were found in the wild-type mice.The total number of intestinal adenoma of Apcmin/+ DOC group significantly increased,compared with that of Apcmin/+ control group (57.00 ± 3.07 vs 21.50± 4.69,t=20.03,P＜0.01),the increase of the adenoma with maximum diameter between 1 to 2 mm was most significant (30.62± 7.73 vs 7.75 ± 4.59,t =8.04,P＜ 0.05),the rate of adenoma canceration also significantly increased compared with that of Apcmin/+ control group.The percentage of PCNA positive cells significantly increased compared with that of Apcmin/+ control group ((90.17 ± 2.14) ％ vs (41.97 ± 4.26) ％,t=31.97,P＜0.01).The percentage of cell apoptosis significantly declined ((1.40± 1.12) ％ vs (7.50 ± 0.65)％,t =14.90,P＜ 0.01).The diversity of intestinal flora of Apcmin/+ DOC group significantly decreased.The ratio of Firmicutes and Bacteroidetes significantly increased compared with control group (0.586 7±0.148 4 vs 0.387 3±0.013 6,t=2.36,P＜0.05).The number of pathogenic bacteria increased in Apcmin/+ DOC group and probiotics significantly decreased.Conclusion DOC can induce intestinal flora imbalance in Apcmin/+ mice and promote intestinal adenoma into adenocarcinoma through increasing tumor cell proliferation and inhibiting cell apoptosis.
