發(fā)布時間：2018-12-11 18:41

【摘要】：目的：以雄激素性脫發(fā)小鼠模型為研究目標(biāo),通過在小鼠背部外用中藥成分齊墩果酸觀察小鼠毛發(fā)生長情況并對其血清T、E2含量、T/E2比例進(jìn)行檢測,探討齊墩果酸對雄激素性脫發(fā)的治療作用及其相關(guān)機(jī)制研究。方法：將56只B6CBAF1/J雄性小鼠按體重編號,按照隨機(jī)數(shù)字表法選出13只小鼠作為空白組,余下小鼠分籠喂養(yǎng)。除空白組外未分組小鼠均按照5mg/Kg·d劑量給予背部皮下注射丙酸睪酮,連續(xù)給藥4周,造成雄激素性脫發(fā)動物模型,空白組注射等量生理鹽水。實(shí)驗(yàn)第4周小鼠造模成功后,在空白組及未分組小鼠中分別隨機(jī)選出3只小鼠頸部脫臼處死,觀察其毛發(fā)脫落情況及脫發(fā)區(qū)皮膚組織病理變化。將剩余的40只未分組小鼠分為模型組、基質(zhì)組、陽性對照組、齊墩果酸組,每組10只小鼠,繼續(xù)給予丙酸睪酮注射,同時齊墩果酸組外用1%齊墩果酸溶液、陽性對照組外用5%米諾地爾溶液、基質(zhì)組外用未加用齊墩果酸的空白溶液,模型組不給藥。第8周實(shí)驗(yàn)結(jié)束時,取小鼠股動脈血液,提取血清。取血后將小鼠頸部脫臼處死,平行脊柱取小鼠背部脫毛區(qū)皮膚組織做HE染色,計(jì)算終毛/毳毛比例。應(yīng)用化學(xué)發(fā)光微粒子免疫分析系統(tǒng)及配套的人睪酮、雌二醇試劑盒測定小鼠血清內(nèi)睪酮T、雌二醇E2的含量及T/E2比例,整個測定過程由儀器按預(yù)定程序自動完成。結(jié)果：1、小鼠造模2周后,背部毛發(fā)逐漸開始失去光澤,造模4周、8周的小鼠,不同程度地出現(xiàn)了背部毛發(fā)脫落,逐漸從頭部向尾部發(fā)展,且脫落加劇；皮膚組織病理提示毛囊微型化,終毛/毳毛數(shù)目比例下降,表明雄激素性脫發(fā)小鼠模型造模成功。2、實(shí)驗(yàn)第8周,小鼠背部皮膚組織HE染色,計(jì)數(shù)終毛與毳毛的數(shù)目,進(jìn)行終毛/毳毛比例比較,齊墩果酸組與模型組、基質(zhì)組比較,比例升高(P0.01)：與空白組比較,比例下降(P0.05)；齊墩果酸組與陽性對照組之間無明顯差異。3、實(shí)驗(yàn)第8周,比較各組小鼠之間血清T、E2含量及T/E2比例。與空白組比較,其余4組小鼠血清內(nèi)睪酮T、雌二醇E2含量、T/E2比例均顯著升高(P0.01),進(jìn)行組間比較,各組T、E2、T/E2無明顯差異(P0.05)。結(jié)論：1、B6CBAF1/J小鼠背部皮下注射丙酸睪酮建立雄激素性脫發(fā)小鼠模型的方法確實(shí)有效,造模效果及模型穩(wěn)定性都較好。2、齊墩果酸治療雄激素性脫發(fā)是有效的,其與陽性對照米諾地爾比較,促進(jìn)毛發(fā)生長的療效相當(dāng)。齊墩果酸在治療雄激素性脫發(fā)的同時還可逆轉(zhuǎn)終毛與毳毛的比例、改善毛囊微型化。3、齊墩果酸外用治療雄激素性脫發(fā)小鼠4周,不能改變小鼠體內(nèi)雄激素T、雌激素E2含量、T/E2比例。
[Abstract]:Objective: to observe the hair growth of mice with androgen induced alopecia by using oleanolic acid (Oleanolic acid) on the back of mice. To investigate the therapeutic effect of oleanolic acid on androgen induced alopecia and its related mechanism. Methods: Fifty-six male B6CBAF1/J mice were selected as blank group according to their body weight number and random number table method. The remaining mice were fed in cage. All the mice except the blank group were subcutaneously injected with testosterone propionate according to the dose of 5mg/Kg d for 4 weeks to make androgen-induced alopecia animal model. The blank group was injected with the same amount of normal saline. At the 4th week of the experiment, 3 mice were randomly selected for neck dislocations and killed in blank group and undivided group. The hair loss and the pathological changes of skin tissue in the alopecia region were observed. The remaining 40 undivided mice were divided into three groups: model group, matrix group, positive control group and oleanolic acid group. 10 mice in each group were given testosterone propionate injection and oleanolic acid group was treated with 1% oleanolic acid solution. The positive control group was treated with 5% minoxidil solution, the matrix group was treated with blank solution without oleanolic acid, and the model group was not treated. At the end of the 8th week, the blood of the femoral artery of mice was taken and the serum was extracted. After blood was taken, the cervical dislocations of the mice were killed, and the skin tissues of the dorsal hair removal area of the mice were taken from the parallel spine for HE staining, and the final hair / vellus hair ratio was calculated. The contents of testosterone, estradiol E _ 2 and T/E2 ratio in serum of mice were determined by chemiluminescence microparticle immunoassay system and matched human testosterone and estradiol kit. Results: 1. After 2 weeks of modeling, the hair on the back began to lose its luster gradually. After 4 weeks and 8 weeks of modeling, the hair loss of the back appeared in varying degrees, and gradually developed from the head to the tail, and the shedding became worse. The skin histopathology indicated that the hair follicles were miniaturized and the number ratio of terminal hair to vellus hair decreased, which indicated that the model of androgen-induced alopecia was successful. 2. At the 8th week of the experiment, HE staining was used to count the number of terminal hair and vellus hair in the skin of mice. The ratio of terminal hair to vellus hair in oleanolic acid group was higher than that in model group and matrix group (P0.01); compared with blank group, the proportion of oleanolic acid group decreased (P0.05). There was no significant difference between the oleanolic acid group and the positive control group. Compared with the blank group, the levels of testosterone, estradiol E _ 2 and T/E2 in the serum of the other four groups were significantly increased (P0.01). There was no significant difference in T _ 2E _ 2 / T / E _ 2 between the four groups (P0.05). Conclusion: 1 the method of subcutaneously injecting testosterone propionate into the back of B6CBAF1 / J mice to establish androgen-induced alopecia mouse model is effective, and the model effect and model stability are good. 2, oleanolic acid is effective in the treatment of androgen-induced alopecia. Compared with the positive control minodil, the effect of promoting hair growth is comparable. Oleanolic acid can also reverse the ratio of terminal hair to vellus hair and improve the miniaturization of hair follicles. Estrogen E 2 content, T/E2 ratio.
【學(xué)位授予單位】：瀘州醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R758.71

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