ALA光动力疗法控制金黄地鼠皮脂腺斑痤疮丙酸杆菌感染后炎症的研究

发布时间：2019-05-16 06:51

【摘要】：背景：多国健康监测系统统计数据显示，人群中约有85％在12～24岁期间出现痤疮，严重影响患者人群的身心健康。痤疮丙酸杆菌(Propi-onibacterium acnes，Pacnes)在痤疮形成中起重要作用，其代谢产物如脂肪酶、蛋白酶、透明质酸酶和趋化因子均可引起炎症反应。皮肤脂质除了为Pacnes大量增殖提供厌氧环境外，其中的一些成分也直接或间接参与了痤疮炎症的发生。光动力疗法(Photod3rnarnic 111er．dpy，PDT)是一种正在研究发展中的新型医疗技术，光动力疗法的优点主要是组织选择性强，对正常组织不会造成损害。5．氨基乙酰丙酸(5．arnin01evulink：acid，，ALA)诱导产生的原卟啉Ix对皮脂腺有很强的选择性。局部用ALA在人体毛囊和皮脂腺中转变成有效的光敏剂(原卟啉Ix)，在细菌中转变成光敏剂(粪卟啉或尿卟啉)，经光激发引起光动力反应，继而抑制皮脂分泌，灭活皮肤角化细胞并杀灭毛囊中的致病菌。目的：本研究旨在观察5氨基乙酰丙酸光动力疗法(5．arnin01eVIIIlinic acid：Photodynarnic T11er．dp)_，ALA-PDT)对Pacnes感染金黄地鼠皮脂腺斑炎症的抑制作用，并进一步探讨ALA．PDT抗炎治疗的作用机理，为皮肤科临床应用ALA PDT治疗痤疮炎症提供实验依据。方法：①取健康金黄地鼠16只，随机分为4组：I组为空白对照组、II组为Pacnes组、III组为Pacnes+ALA．PDT组、Ⅳ组为Pacnes+红光组。厌氧环境下培养痤疮丙酸杆菌并接种于II、III、Ⅳ组金黄地鼠皮脂腺斑，诱导炎症反应产生，1周后进行光动力治疗。②对III组金黄地鼠皮脂腺斑区域先外敷20％．ALA1h后进行红光照射，而Ⅳ组只用红光照射。照光方案为每周1次，共4周。照射参数为：能量密度127 J／cm。波长633±3 nm，光斑大小1 O cm，设定时间为15min。3分别切取各实验组皮脂腺斑进行组织学观察及特殊染色。4酶联免疫吸附测定血清睾酮与雌二醇水平。5westernblot检测炎症信号通路相关蛋白NF．KB和s_lXI3蛋白表达水平。结果： 1．光动力治疗后皮脂腺斑炎症反应减少与P.lcnes组比较，ALA．PDT和单纯红光照射均能减轻痤疮丙酸杆菌感染引起的以炎细胞浸润为主的炎症性改变。ALA．PDT组对皮脂腺斑组织的炎症抑制效果显著强于单纯红光组。 2．光动力治疗对血清睾酮，雌二醇水平的影响与空白对照组比较，ALA．PDT和单纯红光照射均未引起血清睾酮，雌二醇水平的显著差异(PO 05)。 3．光动力治疗对NF—KB和sTAT3蛋白表达水平的影响与P.lcnes组比较，光动力治疗后NF．KB蛋白表达水平明显降低，而s_lXI蛋白表达升高。ALA．PDT组对皮脂腺斑组织炎症信号通路相关蛋白表达抑制效果显著强于单纯红光组，差异均具有统计学意义(PO 05)。结论：光动力疗法通过改变炎症信号通路相关蛋白NF．KB和s_lXI的表达，可对痤疮丙酸杆菌引起的皮脂腺炎症反应有良好的抑制效果。光动力治疗不影响金黄地鼠血清睾酮和雌二醇的表达水平。
[Abstract]:Background: according to the statistics of multinational health monitoring system, about 85% of the population developed acne during the age of 12 years, which seriously affected the physical and mental health of the patients. Propionium acnes (Propi-onibacterium acnes,Pacnes) plays an important role in acne formation. Its metabolites such as lipase, protease, hyaluronidase and chemokines can cause inflammation. In addition to providing an anaerobic environment for the proliferation of Pacnes, some of its components are also directly or indirectly involved in the occurrence of acne inflammation. Photodynamic therapy (Photod3rnarnic 111er.dpy, PDT) is a new medical technology under development. Photodynamic therapy has the advantages of strong tissue selectivity and no damage to normal tissue. 5. 5. Aminolevulinic acid (5.arnin01evulink acid, PDT) is a new medical technology in the process of research and development. the main advantages of photodynamic therapy are that it has strong tissue selectivity and does not cause damage to normal tissue. 5. Aminolevulinic acid, ALA) induced protoporphyrin Ix has strong selectivity to sebaceous gland. Local use of ALA into an effective photosensitizer in human hair follicles and sebaceous gland (protoporphyrin Ix), is transformed into photosensitizer (fecal porphyrin or uroporin) in bacteria, which can induce photodynamic reaction and then inhibit sebum secretion. Inactivate keratinized cells of the skin and kill pathogenic bacteria in hair follicles. Objective: to observe the inhibitory effect of 5-aminolevulinic acid photodynamic therapy (5.arnin01eVIIIlinic acid:Photodynarnic T11er.dp) _ (ALA-PDT) on sebaceous gland plaque inflammation in golden hamsters infected with Pacnes. To further explore the mechanism of anti-inflammatory therapy of ALA.PDT, and to provide experimental basis for clinical application of ALA PDT in the treatment of acne inflammation. Methods: 1 Sixteen healthy golden hamsters were randomly divided into 4 groups: group I was blank control group, II group was Pacnes group, III group was Pacnes ALA.PDT group, group IV was Pacnes red light group. Propionium acnes was cultured in anaerobically environment and inoculated with II,III, IV golden hamster sebaceous gland spot to induce inflammatory reaction. One week later, photodynamic therapy was performed. 2 the area of sebaceous gland spot of golden hamster in III group was irradiated with red light after external application of 20%.ALA1h, while group IV was irradiated with red light only. The lighting scheme is once a week for 4 weeks. The irradiation parameters are as follows: energy density 127 J / cm. Wavelength 633 卤3 nm, spot size 1 O cm, The time was set at 15min.3 for histologic observation and special staining of sebaceous gland plaques in each experimental group. 4 the levels of serum testosterone and estrogen were measured by enzyme-linked immunosorbent assay (Elisa). The inflammatory signaling pathway related proteins NF.KB and Estradiol were detected by Western Blot. The expression level of s_lXI3 protein. Results: 1. Compared with P.lcnes group, the inflammatory reaction of sebaceous plaques decreased after photodynamic therapy. Both ALA.PDT and simple red light irradiation could alleviate the inflammatory changes mainly caused by inflammatory cell infiltration caused by propionic acid bacillus acnes infection. The inhibitory effect of ALA.PDT group on sebaceous ganglia tissue was significantly stronger than that of simple red light group. 2. The effect of photodynamic therapy on serum testosterone and Estradiol levels was not significantly different from that of blank control group (PO 05). There was no significant difference in serum testosterone and estrogen levels between ALA.PDT and red light irradiation alone. 3. The effect of photodynamic therapy on the expression of NF-KB and sTAT3 protein was significantly lower than that of P.lcnes group after photodynamic therapy. However, the expression of s_lXI protein was increased. The inhibitory effect of ALA.PDT group on the expression of inflammatory signal pathway-related proteins in sebaceous ganglia tissue was significantly stronger than that in simple red light group (PO 05), and the effect of ALA.PDT group was significantly higher than that of simple red light group (ALA.PDT group). Conclusion: photodynamic therapy can inhibit the inflammatory reaction of sebaceous gland induced by propionic acid acne by changing the expression of NF.KB and s_lXI. Photodynamic therapy did not affect the expression of serum testosterone and estrogen in golden hamsters.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R758.733

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