本文關(guān)鍵詞：基于中藥方劑配伍機(jī)理的中藥有效成分在食管癌前病變進(jìn)程中的免疫調(diào)節(jié)作用　出處：《河北醫(yī)科大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

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【摘要】：目的:本課題擬研究“連翹、天花粉蛋白、黨參、香加皮”中有效成分組成的“連花參加”方逆轉(zhuǎn)化學(xué)致癌劑4-硝基喹啉-1-氧化物(4-nitro-quinoline-1-oxide,4NQO)誘導(dǎo)小鼠(C57BL/6)食管癌前病變的效果及潛在的免疫調(diào)節(jié)機(jī)制。方法:1選取300只C57BL小鼠動(dòng)物模型,按隨機(jī)原則分為6組,分別為空白對(duì)照組(A1組)(不做任何處理)30只;甲基纖維素對(duì)照組(A2組)30只,給予甲基纖維素灌胃,不給藥;誘癌劑組(B組),治療組(C組),預(yù)防組(D組),全反式維甲酸陽(yáng)性對(duì)照組(E組),用滅菌水配制成100μg/ml濃度,采取正常飲水法給B、C、D、E四組小鼠。受試藥“連花參加”方的配制方法為連翹脂素(2mg):天花粉蛋白(1.4mg):香加皮三萜類化合物(1.4mg):黨參蛋白(0.7mg),按此比例混合溶于1.5%的甲基纖維素,定容20ml。D組在實(shí)驗(yàn)開(kāi)始就給與中藥方劑“連花參加”方灌胃。C組第15周開(kāi)始給予中藥方劑“連花參加”方灌胃,E組也在15周進(jìn)行“全反式維甲酸(ATRA)”灌胃。C、D、E組小鼠每周均灌胃三次,每次0.1ml。2該實(shí)驗(yàn)從第12周時(shí)分別把A1、A2、B、D組小鼠處死5只,觀察結(jié)果顯示小鼠食管未出現(xiàn)不典型增生。在15周時(shí),與12周采取同樣的處理,HE染色結(jié)果顯示小鼠食管發(fā)現(xiàn)早期不典型增生。從15周開(kāi)始,將B組、C組、D組、E組100μg/ml 4NQO水溶液統(tǒng)一更換為滅菌水。第18、21周,A1、A2組各處死5只,B、C、D、E組各處死15只,第24周時(shí),處死剩余小鼠,均為斷頸處死。3建立小鼠癌前病變的模型,HE檢測(cè)B組不同時(shí)期組織的病理學(xué)變化,鑒定腫瘤進(jìn)展的階段。4提取小鼠脾細(xì)胞,觀察用藥前后對(duì)脾細(xì)胞中淋巴細(xì)胞的影響。用流式細(xì)胞法檢測(cè)小鼠脾細(xì)胞中CD4+,CD8+,B7-H4(B7homologue4),CD4+CD25+foxp3+等T細(xì)胞及其表面分子在不同時(shí)間點(diǎn)的組間變化情況。結(jié)果:1成功建立了小鼠食管癌癌前病變的模型,HE染色發(fā)現(xiàn)第15周時(shí)4NQO誘癌組的小鼠開(kāi)始有輕度不典型增生。并且隨著時(shí)間的延長(zhǎng),病變程度逐漸加深。2通過(guò)HE染色結(jié)果顯示,小鼠食管組織病變程度在24周時(shí),有顯著性差異,P0.05,C、D、E組的致癌率比B組顯著降低,P0.05,說(shuō)明“連花參加”方可減緩食管癌病變的進(jìn)程。3實(shí)驗(yàn)結(jié)果顯示在18周和21周時(shí)各組間CD4+CD25+foxp3+調(diào)節(jié)性T細(xì)胞占脾淋巴細(xì)胞的百分率表達(dá)水平無(wú)統(tǒng)計(jì)學(xué)差異;在24周時(shí)各組間有統(tǒng)計(jì)學(xué)差異,P0.05。4 B7-H4在CD4+上的表達(dá)率在21周和24周時(shí)出現(xiàn)顯著性差異,P0.05。B7-H4在CD8+T細(xì)胞和CD4+CD25+foxp3+T細(xì)胞上的表達(dá)率在18周、21周和24周時(shí)均無(wú)顯著性差異。結(jié)論:1成功建立了小鼠食管癌前病變動(dòng)物模型。2“連花參加”方通過(guò)抑制Treg細(xì)胞在脾淋巴細(xì)胞上的表達(dá)和B7-H4在CD4+T細(xì)胞的表達(dá),促進(jìn)了機(jī)體抗腫瘤免疫作用,減緩了食管癌前病變進(jìn)程。
[Abstract]:Objective: to study "Forsythia suspensa, Trichosanthin, Codonopsis pilosula". "Lianhuayan", an active component of Xiangjia Peel, was used to reverse the chemical carcinogen 4-nitro-quinoline-1-oxide (4-nitro-quinoline-1-oxide). (4) the effect of C57BL / 6) on precancerous lesion and its potential immunomodulatory mechanism in mice were induced by NQO. Methods 300 C57BL mice were randomly divided into 6 groups. There were 30 rats in group A 1 (without any treatment) in the blank control group. 30 rats in group A _ 2 of methylcellulose control group were given intragastric administration of methylcellulose without administration of the drug. Cancer inducers group B, treatment group C, prevention group D, all trans retinoic acid positive control group, group E, 100 渭 g / ml of sterilizing water were given to B by normal drinking water. C,D. Group E mice. The preparation method of "Lianhua participating" prescription was Forsythia suspensa 2mg / g: Trichosanthin 1.4 mg / g: trichosanthin = 1.4 mg / g: Codonopsis Codonopsis protein (1. 4 mg / g / L); trichosanthin 1. 4 mg / g; triterpenoid compound 1. 4 mg / g; (0.7 mg). In this proportion, methyl cellulose dissolved in 1.5% was mixed. At the beginning of the experiment, group D was given the traditional Chinese medicine prescription "continuous flower participation". Group C was given the traditional Chinese medicine prescription "continuous flower participation" by stomach at the 15th week. Group E was also given ATRA-ATRA for 15 weeks. Mice in group E were given oral administration of A1A _ 2 three times a week, 0.1ml.2 each time from the 12th week. Five mice in group B were killed. The results showed that there was no atypical hyperplasia in the esophagus of the mice, and the same treatment was taken at 15 weeks and 12 weeks. The results of HE staining showed that early atypical hyperplasia was found in the esophagus of mice. From 15 weeks on, 100 渭 g / ml 4NQO solution of group B and C was replaced with 100 渭 g / ml 4NQO solution as sterilizing water. In group A _ 1 A _ 2, 15 rats were killed everywhere in group A _ 1 A _ 2 at 21 weeks. At the 24th week, the remaining mice were killed, all of them were killed by cervical amputation to establish the model of precancerous lesion in mice. He was used to detect the pathological changes of the tissues in group B at different stages and to identify the stage of tumor progression. 4. The spleen cells were extracted from mice. To observe the effect on lymphocytes in splenocytes before and after treatment, and to detect the CD4 CD8 B7-H4 B7homologue 4 in spleen cells of mice by flow cytometry (FCM). The changes of T cells and their surface molecules such as CD4 CD25 foxp3 were observed at different time points. Results the mouse model of precancerous lesion of esophageal carcinoma was successfully established at 1: 1. He staining showed that the mice in 4NQO induced cancer group began to have mild atypical hyperplasia at the 15th week, and with the extension of time, the degree of lesion gradually deepened by HE staining results showed. At 24 weeks after 24 weeks, there was significant difference in carcinogenic rate of P0.05CU DU E group compared with B group (P0.05). The results show that "Lianhua attends" can slow down the progression of esophageal cancer. 3. 3. The results show that CD4 CD25 foxp3 can be found between groups at 18 and 21 weeks. There was no significant difference in the percentage of regulatory T cells in spleen lymphocytes. There was a significant difference in the expression rate of P0.05.4 B7-H4 on CD4 between the groups at 24 weeks and 21 weeks and 24 weeks. The expression rate of P0.05.B7-H4 on CD8 T cells and CD4 CD25 foxp3 T cells was 18 weeks. There was no significant difference between 21 weeks and 24 weeks. Conclusion the mouse model of precancerous lesion was successfully established by 1: 1. The expression of Treg cells on spleen lymphocytes and B7-H4 on CD4 T cells were inhibited. Promoted the body anti-tumor immunity, slowed down the process of esophageal precancerous lesions.
【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.1

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相關(guān)期刊論文 前1條

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