發(fā)布時間：2018-04-30 13:42

  本文選題：原發(fā)性肝癌 + 5-氟尿嘧啶��； 參考：《河北醫(yī)科大學》2017年碩士論文

【摘要】：目的:本研究以癌株傳代的方式建立H22荷瘤小鼠模型,通過觀察化濁解毒固本方、5-氟尿嘧啶(5-Fluorouridine,5Fu)以及化濁解毒固本方聯(lián)合5-氟尿嘧啶等不同治療干預對H22荷瘤小鼠生存狀態(tài)、瘤體大小、免疫情況、病理變化以及相關(guān)腫瘤指標的影響,評價聯(lián)合用藥的抗癌效果,從肝癌的治療機理入手,探討中醫(yī)藥在肝癌的中西醫(yī)結(jié)合治療中所發(fā)揮的作用,為原發(fā)性肝癌的中西醫(yī)結(jié)合治療提供實驗依據(jù)。方法:通過癌株傳代法將稀釋后的腫瘤細胞注射于小鼠右腋皮下,建立H22肝癌小鼠模型。將已成功生長腫瘤的40只小鼠隨機分為4組,每組10只,包括模型組(M)、化濁解毒固本方組(Z)、5-氟尿嘧啶組(X)以及化濁解毒固本方聯(lián)合5-氟尿嘧啶組(L)。造模成功后給予相應藥物進行干預,其中中藥組以化濁解毒固本方配方?jīng)_劑0.2ml/d劑量灌胃治療;西藥組以5-氟尿嘧啶20mg/kg/d腹腔注射治療;聯(lián)合用藥組上午給予化濁解毒固本方灌胃治療,下午予5-氟尿嘧啶腹腔注射治療;模型組以0.4ml/d生理鹽水進行灌胃維持。連續(xù)給藥12天,定期觀察各組小鼠的毛色光澤、進食情況、精神狀態(tài)、體重變化以及死亡情況等,于末次治療后次日,頸椎脫臼法處死小鼠,并鈍性剝離小鼠腫瘤組織、脾臟組織、胸腺組織,分別計量稱重,計算各治療組的抑瘤率、脾指數(shù)、胸腺指數(shù)并進行統(tǒng)計分析,對腫瘤組織進行甲醛固定、病理切片,采用免疫組化法(Immunohistochmeistry,IHC)檢測VEGF、MMP-9蛋白的表達,并對蘇木素-伊紅(Hematoxyli/eosin,HE)染色后的腫瘤組織進行病理觀察。結(jié)果:1小鼠的一般狀況各實驗組小鼠治療后生存狀態(tài)都明顯不如治療前,尤其模型組,小鼠精神萎靡,毛色枯糙,食欲不振,體型瘦削,體重無明顯增長,右腋下腫物明顯腫大可見。與西藥組相比,中藥組和聯(lián)合用藥組在精神狀態(tài)、食欲情況和體重增長等方面都占明顯優(yōu)勢。2小鼠的腫瘤生長情況在抑瘤方面,聯(lián)合用藥組抑瘤率最高,西藥組抑瘤率高于中藥組。平均瘤重方面,聯(lián)合用藥組瘤重最小,其次是西藥組、中藥組、模型組。三個治療組瘤重均小于模型組,差異有統(tǒng)計學意義(p0.05);其中,聯(lián)合用藥組瘤重小于西藥組和中藥組,差異顯著(p0.05);西藥組瘤重小于中藥組,差異有統(tǒng)計學意義(p0.05)。3小鼠脾指數(shù)情況中藥組小鼠脾指數(shù)最高,明顯高于其它各組,差異具有統(tǒng)計學意義(p0.05);聯(lián)合用藥組次之,明顯高于西藥組和模型組,差異明顯(p0.05);西藥組小鼠脾指數(shù)較之模型組,差異無統(tǒng)計學意義(p0.05)。4小鼠胸腺指數(shù)情況與模型組相比,各治療組荷瘤小鼠胸腺指數(shù)均顯著升高,差異有統(tǒng)計學意義(p0.05);肝癌小鼠各治療組間相比,中藥組、聯(lián)合用藥組小鼠胸腺指數(shù)明顯高于西藥組,差異明顯(p0.05);聯(lián)合用藥組胸腺指數(shù)與中藥組相比,差異無統(tǒng)計學意義(p0.05)。5小鼠腫瘤組織中vegf的表達各治療組肝癌小鼠腫瘤組織中vegf含量明顯低于模型組,差異有統(tǒng)計學意義(p0.05);聯(lián)合用藥組腫瘤組織中vegf含量顯著低于中藥組和西藥組,差異有統(tǒng)計學意義(p0.05);西藥組腫瘤組織中vegf含量低于中藥組,差異有統(tǒng)計學意義(p0.05)。6小鼠腫瘤組織中mmp-9的表達肝癌小鼠各組間比較,腫瘤組織中mmp-9蛋白表達量有顯著差異,與模型組相比,各治療組組織mmp-9蛋白表達量均明顯降低,差異具有統(tǒng)計學意義(p0.05);其中聯(lián)合用藥組mmp-9表達量最低,明顯低于中藥組和西藥組,差異顯著(p0.05);西藥組組織mmp-9低于中藥組,差異有統(tǒng)計學意義(p0.05)。7小鼠腫瘤組病理變化情況模型組腫瘤組織生長旺盛,腫瘤細胞特異性強,核染顏色很深,嗜堿性很強,鏡下血管豐富;中藥組腫瘤組織生長緩慢,大量腫瘤細胞出現(xiàn)變性裂解,組織出現(xiàn)大面積壞死,細胞質(zhì)細胞核嗜酸性增強;西藥組腫瘤組織生長不旺盛,明顯出現(xiàn)片狀腫瘤壞死區(qū)和空泡,細胞異型性降低,細胞核呈現(xiàn)弱酸性;聯(lián)合用藥組腫瘤組織生長不明顯,部分腫瘤細胞發(fā)生壞死溶解,細胞質(zhì)細胞核嗜酸性很強,鏡下血管較模型組明顯減少。,結(jié)論:1化濁解毒固本方能夠增強5-氟尿嘧啶的抑瘤作用,提高腫瘤抑制率,其作用機理可能與抑制腫瘤血管生成有關(guān)。2化濁解毒固本方能夠明顯改善5-氟尿嘧啶對H22肝癌小鼠的免疫抑制作用,增強肝癌小鼠的免疫功能,提高肝癌小鼠的生存質(zhì)量。
[Abstract]:Objective: to establish a H22 tumor bearing mouse model in this study, the survival state of the tumor bearing mice, the size of the tumor, the immune situation, the pathological changes and the related tumor indexes were observed by observing the different treatments of the H22, 5- fluorouracil (5-Fluorouridine, 5Fu) and the combined treatment of the turbid and detoxifying decoction. To evaluate the anti-cancer effect of combined use of the drug, from the treatment mechanism of liver cancer, to explore the role of traditional Chinese medicine in the combination of traditional Chinese and Western medicine in the treatment of liver cancer, and to provide experimental basis for the combination of traditional Chinese and Western medicine for primary liver cancer. 40 mice with successful tumor growth were divided into 4 groups randomly, with 10 rats in each group, including model group (M), turbidity detoxification solid recipe group (Z), 5- fluorouracil group (X) and combined 5- fluorouracil group (L). The dose of 0.2ml/d was treated with dose of gavage; the western medicine group was treated by intraperitoneal injection of 5- fluorouracil 20mg/kg/d; the combined drug group was given the gavage in the morning, and the 5- fluorouracil was administered intraperitoneally in the afternoon. The model group was administered with 0.4ml/d physiological saline for 12 days, and the hair color luster was observed regularly in each group. Eating condition, mental state, body weight change and death condition, the next day after the last treatment, the mice were killed by dislocated cervical vertebra, and the tumor tissue, spleen tissue and thymus tissue of the mice were weighed blunt, and the tumor suppressor rate, the spleen index, the thymus index and the statistical analysis were calculated. Immunohistochmeistry (IHC) was used to detect the expression of VEGF, MMP-9 protein, and the pathological observation of the tumor tissue after hematoxylin eosin (Hematoxyli/eosin, HE) staining. Results: the general condition of 1 mice was obviously inferior to that of the mice after treatment, especially in the model group, and in mice. The mental state, appetite and body weight growth of the Chinese medicine group and the combination group are all obviously superior to the tumor growth of.2 mice, and the combination group has the highest tumor suppressor rate, and the combination group has the highest tumor suppressor rate. The tumor weight of the western medicine group was higher than that of the traditional Chinese medicine group. The tumor weight of the combined drug group was the smallest, followed by the western medicine group, the traditional Chinese medicine group and the model group. The tumor weight of the three treatment groups was less than the model group, the difference was statistically significant (P0.05); the tumor weight of the combined drug group was less than the western medicine group and the traditional Chinese medicine group, the difference was significant (P0.05); the tumor weight of the western medicine group was less than the Chinese traditional medicine group. The difference was statistically significant (P0.05).3 mice spleen index, the spleen index of the Chinese medicine group was the highest, obviously higher than the other groups, the difference was statistically significant (P0.05); the combination of drug group was significantly higher than the western medicine group and the model group, the difference was obvious (P0.05); the difference of spleen index in the western medicine group was less than that of the model group (P0.05).4 small Compared with the model group, the thymus index of the mice was significantly higher than that in the model group, and the difference was statistically significant (P0.05). The thymus index of the mice in the Chinese medicine group and the combination group was significantly higher than that in the western medicine group (P0.05), and the thymus index of the combined drug group had no difference compared with the Chinese medicine group. The expression of VEGF in the tumor tissue of.5 mice was significantly lower than that in the model group. The difference was statistically significant (P0.05), and the content of VEGF in the tumor tissue of the combined drug group was significantly lower than that of the traditional Chinese medicine group and the western medicine group (P0.05); VEGF contained in the tumor tissue of the western medicine group contained VEGF content (P0.05). The difference was significantly lower than the Chinese medicine group. The difference was statistically significant (P0.05) in the tumor tissue of.6 mice, the expression of MMP-9 protein in the tumor tissues was significantly different. Compared with the model group, the expression of MMP-9 protein in each group was significantly decreased, and the difference was statistically significant (P0.05), and the combination group mmp- was statistically significant (P0.05), and mmp- in the combination group mmp- was statistically significant (mmp-). The expression of 9 was the lowest, obviously lower than that of the traditional Chinese medicine group and the western medicine group (P0.05). The MMP-9 in the western medicine group was lower than the traditional Chinese medicine group. The difference was statistically significant (P0.05) the pathological changes in the tumor group of.7 mice were strong, the tumor cells were strong, the color of the nuclear dye was very strong, the blood vessels were rich under the microscope, and the swelling of the traditional Chinese medicine group was swollen. The tumor tissue grew slowly, a large number of tumor cells appeared denatured and cracked, tissue appeared large area necrosis and cytoplasm nucleus eosinophilia increased, and the tumor tissue in the western medicine group was not strong, the tumor necrosis area and vacuoles appeared obviously, the cell isomorphism decreased and the nucleus showed weak acidity, and the tumor tissue of the combined drug group was not obvious, part of the tumor tissue was not obvious. The tumor cells were necrotic and dissolved, cytoplasmic cell nuclear eosinophilia was very strong, and the blood vessels under the microscope decreased obviously. Conclusion: 1 the anti tumor effect of 5- fluorouracil can be enhanced and the tumor suppressor rate is enhanced. The mechanism of the action may be related to the inhibition of the angiogenesis of the tumor angiogenesis with the inhibition of the tumor angiogenesis in the 5- fluoruria. The immunosuppressive effect of pyrimidine on H22 hepatoma mice can enhance the immune function of liver cancer mice and improve the quality of life of liver cancer mice.

【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：R735.7

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