本文關(guān)鍵詞：腎虛型慢性再生障礙性貧血小鼠模型建立　出處：《中國中西醫(yī)結(jié)合雜志》2017年09期 　論文類型：期刊論文

  更多相關(guān)文章： 腎虛型 慢性再生障礙性貧血 模型制備

【摘要】：目的建立腎虛型慢性再生障礙性貧血(chronic aplastic anemia,CAA)小鼠模型。方法將70只小鼠行體能測試,篩選出體能相近的小鼠隨機分成腎陰虛組(20只)、腎陽虛組(20只)及正常組(10只),其余隨機分成苯組(10只)及玉米油組(10只)。腎陰虛組每日灌胃甲狀腺素片水溶液(350 mg/kg),共21天;腎陽虛組每日皮下注射氫化可的松(25 mg/kg),共14天。正常組皮下注射生理鹽水(25 mg/kg),共14天,并灌胃生理鹽水(350 mg/kg),共21天。在腎虛模型建立的基礎(chǔ)上,給予皮下注射苯試劑和玉米油混合液2 m L/kg,每周3次,共25次,建立腎虛型CAA小鼠模型。相應(yīng)藥物介入1 d起,觀察小鼠日常生活狀態(tài),通過體能測試監(jiān)測游泳時間;采用ELISA試劑盒測定血清T3、T4、尿17-羥皮質(zhì)類固醇(17-hydroxy-cortico steroid,17-OHCS)、環(huán)磷酸腺苷(c AMP)、環(huán)磷酸腺苷/環(huán)磷酸鳥苷(c AMP/c GMP)水平;血細胞分析儀檢測白細胞、血紅蛋白及血小板數(shù)量;電子顯微鏡下計數(shù)骨髓有核細胞;HE染色觀察骨髓病理特征。結(jié)果藥物干預(yù)25 d后,與正常組比較,腎陰虛組游泳耗時延長、腎陽虛組游泳耗時縮短(P0.01)。藥物干預(yù)22 d后,腎陰虛組小鼠T3、T4、17-OHCS、c AMP、c AMP/c GMP水平上升;腎陽虛組T3、T4、17-OHCS、c AMP/c GMP水平下降(P0.05,P0.01)。苯試劑介入第12次時,與正常組比較,腎陰虛+苯組、腎陽虛+苯組及苯組白細胞、血小板計數(shù)降低(P0.05,P0.01)。苯試劑介入第18次時,與正常組比較,腎陰虛+苯組、腎陽虛+苯組及苯組白細胞、血紅蛋白、血小板計數(shù)均下降(P0.05,P0.01)。藥物介入75 d,腎陰虛+苯組、腎陽虛+苯組及苯組骨髓有核細胞數(shù)均下降(P0.05)。藥物介入75 d,苯試劑導(dǎo)致骨髓增生明顯低下,血白細胞、血紅蛋白及血小板計數(shù)均有不同程度減少,非造血細胞比例增高。結(jié)論灌胃甲狀腺素法聯(lián)合皮下注射苯試劑法可成功建立腎陰虛型CAA小鼠模型;皮下注射氫化可的松法聯(lián)合皮下注射苯試劑法可成功建立腎陽虛型CAA小鼠模型。
[Abstract]:Objective to establish a mouse model of chronic aplastic anemia (CAA) with kidney deficiency. Methods a total of 70 mice were selected for physical performance test, and the mice with similar physical strength were randomly divided into 20 groups: kidney yin deficiency group (20 cases), kidney yang deficiency group (20 cases) and normal group (10). The rest were randomly divided into benzene group (n = 10) and corn oil group (10 rats). The kidney yin deficiency group was injected with water solution of thyroxine tablets (350 mg/kg) daily for 21 days, and the hypodermic injection of hydrocortisone (25 mg/kg) daily for 14 days in the kidney yang deficiency group. Normal saline (25 mg/kg) was subcutaneously injected into the normal group for 14 days, and the saline (350 mg/kg) was administered to the stomach (350 mg/kg), for a total of 21 days. On the basis of the establishment of kidney deficiency model, subcutaneous injection of benzene reagents and corn oil mixture 2 m L/kg, 3 times a week, a total of 25 times, the establishment of kidney deficiency type CAA mice model. The corresponding drug intervention 1 D, observed the state of daily life, through the physical fitness test monitoring swimming time; serum T3, T4, 17- in urine hydroxycorticosteroid using ELISA Kit (17-hydroxy-cortico steroid, 17-OHCS), cyclic adenosine monophosphate (C AMP), cyclic adenosine monophosphate / cyclic guanosine monophosphate (C AMP/c GMP); the number of blood cell analyzer detection of white blood cell, hemoglobin and platelet count under electron microscope; bone marrow cell; bone marrow pathology HE staining. Results after 25 D, compared with the normal group, the swimming time of the kidney yin deficiency group was prolonged and the time of swimming in the kidney yang deficiency group was shortened (P0.01). After 22 D intervention, the levels of T3, T4, 17-OHCS, C AMP and C AMP/c GMP increased in the kidney yin deficiency group, while the levels of T3, T4, GMP, and gamma in the kidney yang deficiency group decreased. Compared with the normal group, the kidney yin deficiency + benzene group, kidney yang deficiency + benzene group and benzene group white blood cell and platelet count decreased (P0.05, P0.01) compared with the normal group for Twelfth times. When benzene reagent was intervened eighteenth times, compared with the normal group, the kidney yin deficiency + benzene group, kidney yang deficiency + benzene group and benzene group white blood cell, hemoglobin and platelet count decreased (P0.05, P0.01). Drug intervention 75 D, kidney yin deficiency + benzene group, kidney yang deficiency + benzene group and the number of nucleated cells in the benzene group were decreased (P0.05). Drug intervention for 75 D, benzene reagents caused bone marrow hyperplasia was significantly lower, white blood cells, hemoglobin and platelet count decreased to varying degrees, the proportion of non hematopoietic cells increased. Conclusion the CAA mice model of kidney yin deficiency can be successfully established by intragastric administration of thyroxine combined with subcutaneous injection of benzene reagent. Subcutaneous injection of hydrocortisone combined with subcutaneous injection of benzene reagent can successfully establish a CAA mouse model of kidney yang deficiency.
【作者單位】： 浙江大學醫(yī)學院附屬第一醫(yī)院急診科;杭州市余杭區(qū)中醫(yī)院腫瘤內(nèi)科;浙江中醫(yī)藥大學第一臨床醫(yī)學院;浙江省麗水市中醫(yī)醫(yī)院;浙江中醫(yī)藥大學動物實驗研究中心;浙江中醫(yī)藥大學附屬第一醫(yī)院血液科;
【基金】：國家自然科學基金資助項目(No.81373634) 浙江省高等教育質(zhì)量工程-中青年學科帶頭人資助項目(No.GK2011,No.GK2012)
【分類號】：R259;R-332
【正文快照】： 慢性再生障礙性貧血(chronic aplastic anemia,CAA)具有病情反復(fù)、療程較長、可緩解卻難于治愈等特點,中醫(yī)學將其納入“虛勞”范疇,根據(jù)臨床表現(xiàn),通常分為腎陰虛、腎陽虛、腎陰陽兩虛3型。中醫(yī)藥在治療上以補腎為本,輔以健脾、活血化瘀、清熱解毒等法,能明顯增強機體免疫功能,

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