本文選題：神經(jīng)生長因子　切入點：潰瘍性結(jié)腸炎　出處：《蘭州大學(xué)》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：目的使用神經(jīng)生長因子(Nerve growth factor,NGF)對葡聚糖硫酸鈉(Dextran sulfate sodium,DSS)誘導(dǎo)的急性潰瘍性結(jié)腸炎小鼠進(jìn)行藥物干預(yù),探討神經(jīng)生長因子(NGF)對小鼠潰瘍性結(jié)腸炎的作用及可能機(jī)制。 方法16只6-8周齡(18-20g)的雌性SPF級Balb/c小鼠自由飲用5%DSS蒸餾水溶液7天后繼續(xù)飲用蒸餾水7天,ELISA法測定腸組織NGF含量隨時間變化。將40只Balb/c小鼠,按照隨機(jī)原則分為A,B,C,D四組,每組10只,設(shè)：A組,正常對照組； B組,模型組,又叫DSS組； C組,DSS+Ad-NGF組,簡稱Ad-NGF組; D組,DSS+Ad-0組,簡稱Ad-0組。除A組飲用蒸餾水外,其余三組小鼠自由飲用5%DSS蒸餾水溶液,進(jìn)食標(biāo)準(zhǔn)飼料。第7天,四組小鼠禁食12小時,不限飲水,第8天,A,B組動物經(jīng)肛灌注0.9%生理鹽水100μ L-次,C組經(jīng)肛注入100μ L含1.O×109CFU/mL Ad-NGF, D組注入100u L1.0x109CFU/mL, B, C, D組飲用的5%DSS蒸餾水溶液改為正常飲水,一直持續(xù)到第14天。每天定時記錄各組小鼠的一般狀態(tài),大便性狀,隱血／肉眼血便,行疾病活動度(DAI)評分,第14天脫頸處死小鼠,采集結(jié)腸組織,測量結(jié)腸長度,部分組織石蠟包埋,行HE染色和免疫組化染色,部分結(jié)腸標(biāo)本放置于-80℃冰箱保存,比色法及ELISA法測定MPO及IL-10,IL-6,TNF-a,免疫印跡法測定CGRP變化。 結(jié)果結(jié)腸組織NGF的濃度從第1天到第5天逐漸增加(0.095±0.032pg/g,0.32+0.24pg/g結(jié)腸組織),第6天達(dá)到最高值(0.52±0.45pg/g結(jié)腸組織),然后從第7天開始逐漸從0.09±0.04pg/g結(jié)腸組織下降到對照水平；而在第10天,濃度(0.06±0.023pg/g)較對照組低。DAI和組織學(xué)評分在DSS組顯著升高(P0.01),但在Ad-NGF直腸內(nèi)給藥后明顯下降(P0.05),而AD-0組該數(shù)據(jù)無明顯變化。與Ad-0組(9.15±0.21cm)相比, NGF能夠減弱DSS致(10.33+0.36cm)結(jié)腸長度的縮短(對照組vsDSS組：12.67±0.38cmvs7.98±0.15cm,p0.01)。同樣,NGF顯著抑制了DSS誘導(dǎo)的炎性因子IL-6(DSS組vs Ad-NGF:304.57±3.19vs56.67±5.89, p0.05), TNF-a (115.89±6.91,69.14＝4.48, p0.05)的升高,但促進(jìn)了IL-10的分泌(65.34±6.44,160.73±6.42,p0.05),Ad-0無此作用。NGF顯著抑制了DSS誘導(dǎo)的結(jié)腸炎模型中的MPO濃度的增高,Ad-NGF組濃度低于DSS組和Ad-0組(p0.05),DSS組和Ad-0組濃度無明顯差異。DSS顯著抑制了腸組織CGRP的產(chǎn)生,而；補(bǔ)充NGF顯著升高了CGRP的含量,Ad-0無此作用。 結(jié)論神經(jīng)生長因子可減輕DSS誘導(dǎo)的小鼠結(jié)腸炎性損傷；抑制MPO和炎性細(xì)胞因子,促進(jìn)IL-10和CGRP的分泌可能是其抗炎的機(jī)制。神經(jīng)生長因子可能成為臨床治療潰瘍性結(jié)腸炎一個新的選擇。
[Abstract]:Objective to investigate the effect of nerve growth factor (NGF) on ulcerative colitis in mice induced by Dextran sulfate sodiumn (DSS) and its possible mechanism. Methods Sixteen female SPF grade Balb/c mice (6-8 weeks old) were randomly divided into four groups: 5%DSS distilled water solution for 7 days and then distilled water for 7 days. 40 Balb/c mice were randomly divided into four groups. Ten rats in each group were divided into two groups: group A, normal control group; group B, model group, also called DSS group; group C, DSS Ad-NGF group, referred to as Ad-NGF group; group D, DSS Ad-0 group, referred to as Ad-0 group. Except for group A, the other three groups of mice drank 5%DSS distilled water solution freely. On the 7th day, the mice in the four groups fasted for 12 hours, without restriction on drinking water. On the 8th day, 100 渭 L of 0.9% normal saline was injected into the anus of rats in group B and 100 渭 L of 109CFU/mL Ad-NGF was injected into the anus of group C, and the 5%DSS distilled water solution of group D was injected with 100u L 1.0 x 109 CFU / mL, B, C, D instead of normal drinking water. It lasted until the 14th day. The general state, defecation character, occult blood / naked blood stool, disease activity and DAI score were recorded regularly every day. The mice were killed by removing neck on the 14th day, the colonic tissue was collected and the colonic length was measured. Some tissues were embedded in paraffin wax and stained with HE and immunohistochemistry. Some colon specimens were stored in refrigerator at -80 鈩，

本文編號：1652378