驻极体促进胰岛素经皮吸收及其机制的研究

发布时间：2018-06-12 21:46

本文选题：驻极体 + 胰岛素　；参考：《第二军医大学》2015年硕士论文

【摘要】：糖尿病是以慢性高血糖为症状的自身代谢紊乱性疾病,已成为继心脑血管疾病和肿瘤之后危害人类健康的“第三杀手”。目前针对糖尿病患者的治疗药物主要有胰岛素和非胰岛素类药物,胰岛素是当前糖尿病病人控制血糖最常用的药物,临床上通常采用皮下注射的方式给药以获得有效的降糖效果。但是,皮下注射不仅因疼痛不适等缺点给患者带来不便和痛苦,导致患者的依从性差,而且长期频繁注射还可引起患者皮下脂肪萎缩和低血糖等不良反应。为此,非侵入性给药方式,如胰岛素经皮给药制剂成为研究的热点,它可有效避免药物经胃肠道及肝脏的首过效应,提高患者用药的顺应性,减轻患者的痛苦。因此,研究胰岛素经皮给药具有非常重要的意义。由于皮肤最外层角质层的屏障作用,大多数药物的经皮转运速度或程度都很小或有限,需要各种促渗方法的推动。课题组前期的研究结果表明,驻极体对水杨酸甲酯、利多卡因、美洛昔康和环孢菌素A等药物具有显著的经皮促渗作用。因此,课题组在前期研究的基础上,采用驻极体、化学促渗剂和驻极体与化学法促渗剂联合等方法对胰岛素的经皮给药开展研究,探讨驻极体促进胰岛素经皮吸收的可行性及其相关机制,为探索驻极体促进多肽类药物的经皮吸收研究提供-种新方法和新思路。本课题从以下几个方面开展了研究。(一)驻极体对胰岛素的极化及其规律的研究应用常温栅控恒压电晕充电系统制备了不同极性、不同表面电位的驻极体,作用于胰岛素溶液和贴剂,测量施加在胰岛素溶液和胰岛素贴剂两侧的电场强度和压电系数d33随时间的变化规律,考察驻极体对胰岛素的极化及其规律。结果表明：(1)驻极体能够为胰岛素溶液及胰岛素贴剂提供稳定的静电场。(2)驻极体能够引起胰岛素的极化,经不同极性和不同表面电位驻极体作用12小时,胰岛素的极化趋于饱和(即d33值趋于稳定)。(3)驻极体的表面电位越大,引起胰岛素的极化越明显。同时应用核磁共振、傅里叶红外吸收光谱和非变性凝胶电泳技术,进一步研究了驻极体对胰岛素分子结构的影响。结果显示胰岛素经驻极体作用后,其分子的空间结构没有发生明显改变,但分子的部分微结构发生细微变化,是驻极体电场引起胰岛素分子极化的结果。(二)驻极体、化学促渗剂及驻极体和化学促渗剂联用对胰岛素体外透皮规律的影响借助Franz扩散池和高效液相实验技术,系统考察了3%氮酮、7.5%丙二醇化学促渗剂,±500V、±1000V、±1500V驻极体,±1500V驻极体联用3%氮酮对胰岛素的经皮促渗作用,以及胰岛素在角质层和去角质层皮肤中的滞留量。结果显示,(1)氮酮和丙二醇对胰岛素具有较好的促渗作用,且3%氮酮的促渗效果明显优于7.5%丙二醇。化学促渗剂作用胰岛素体外透皮36h后,3%氮酮组的胰岛素在角质层和去角质层皮肤中的分布量最大。(2)负极性驻极体对胰岛素具有良好的体外促渗效果,且驻极体表面电位绝对值越高,对胰岛素的促渗效果越好。驻极体作用后,胰岛素在角质层中的分布与对照组相比无明显区别,但是在去角质层皮肤中的分布明显增加。(3)-1500V驻极体联合3%氮酮,36小时后胰岛素的累积透皮量分别是3%氮酮组、+1500V驻极体联合3%氮酮组和对照组的1.4倍、2.12倍和2.93倍。-1500V驻极体与3%氮酮联合对胰岛素具有很好的协同促渗作用。(三)驻极体对胰岛素经糖尿病大鼠皮肤的体外透皮规律研究本文通过注射链脲佐菌素成功构建了糖尿病大鼠模型,考察了胰岛素溶液、经-1500V、-1000V、-500V驻极体作用后的胰岛素溶液对糖尿病大鼠皮肤的体外透皮规律。结果显示,胰岛素溶液经糖尿病大鼠皮肤的透过量显著高于正常皮肤的透过量,36小时的累积透过量是经正常皮肤透过量的3.2倍。36小时后,经-1500V、-1000V、-500V驻极体作用后的胰岛素溶液对糖尿病大鼠皮肤的累积透过量是经正常皮肤透过量的4.00、3.86、3.54倍,表面电位绝对值越高,提高量越多。(四)驻极体对正常大鼠和糖尿病皮肤显微结构的影响研究利用免疫荧光技术和显微结构分析技术,研究了不同极性、不同表面电位驻极体作用正常和糖尿病大鼠的显微结构。结果显示1)驻极体可使正常大鼠皮肤的角质层变薄,细胞间隙增大,毛囊口扩大。2)驻极体作用糖尿病大鼠皮肤后,皮肤的角质层及真皮层略微变厚,对其有修复作用。3)驻极体作用后的胰岛素其主要经皮途径是皮肤的毛囊结构。(五)驻极体对胰岛素降糖作用的影响经驻极体作用的胰岛素溶液与未经驻极体作用的胰岛素溶液具有相似的降糖规律,经+1000V驻极体和-1000V驻极体作用胰岛素的降糖效果更佳。驻极体胰岛素经皮给药系统具有稳定性好,使用方便,生物安全性好和不需外置仪器设备等特点。驻极体胰岛素透皮给药系统有望糖尿病患者提供新型的治疗方法和手段。
[Abstract]:Diabetes is a metabolic disorder characterized by chronic hyperglycemia, which has become the "third killer" of human health after cardiovascular disease and tumor. At present, the main treatment drugs for diabetic patients are insulin and non insulin, and insulin is the most commonly used diabetic patients to control blood sugar. Medications are usually administered by subcutaneous injection to obtain effective hypoglycemic effect. However, subcutaneous injection is not only inconvenient and painful to patients because of pain discomfort, which leads to poor compliance of the patient, and long-term frequent injections can also cause adverse reactions such as hypodermic atrophy and hypoglycemia. The way of sex administration, such as insulin transdermal drug delivery, has become a hot spot of research. It can effectively avoid the first effect of the drugs through the stomach and the intestines and the liver, improve the compliance of the patients and alleviate the sufferings of the patients. Therefore, it is of great significance to study the transdermal delivery of insulin. The speed or degree of transdermal delivery of drugs is very small or limited, which requires the promotion of various methods of stimulation. The results of the previous research group showed that the electret had significant transdermal enhancers to Methylis salicylas, lidocaine, meloxicam and cyclosporin A. In order to explore the feasibility and related mechanism of electret promoting insulin absorption by the combination of chemical stimulating agent and electret and chemical permeating agent, the feasibility and mechanism of electret to promote the absorption of insulin by the electret are explored. Study. (1) study on polarization and regularity of insulin by electret application of constant temperature gate controlled constant pressure corona charging system to prepare electret with different polarity, different surface potential, acting on insulin solution and patch, measuring the electric field intensity and pressure coefficient d33 applied on both sides of insulin solution and insulin patch with time The polarization of the electret to insulin and its regularity were investigated. The results showed that: (1) the electret could provide stable electrostatic field for insulin solution and insulin patch. (2) the electret could cause insulin polarization, and the polarization of insulin was saturated by different polarity and different surface potential electret (d33). The higher the surface potential of the electret is (3) the greater the surface potential of the electret, the more obvious the polarization of insulin. At the same time, the effect of the electret on the molecular structure of insulin is further studied by NMR, Fourier infrared absorption spectroscopy and non denatured gel electrophoresis. The results show that the structure of the molecules is not in the space of the insulin after the action of the electret. There was obvious change, but the partial microstructure of the molecule changed slightly, which was the result of insulin molecular polarization induced by electret electric field. (two) the influence of electret, chemical permeation agent, electret and chemical permeation agent on the transdermal regulation of insulin in vitro was systematically investigated by Franz diffusion pool and high performance liquid phase experiment. 3% azone was systematically investigated. 7.5% propanediol chemical enhancer, + 500V, + 1000V, + 1500V electret, 3% azone combined with Azone on the skin of insulin, and the retention of insulin in the cuticle and the skin of the cuticle. The results showed that (1) azone and propanediol had better infiltration effect on insulin, and the effect of Azone was obviously better than that of 3% azone. The distribution of insulin in the 3% azone group is the largest in the cuticle and the cuticle skin after the action of 7.5% propanediol. (2) the negative electret has a good exosperation effect on the insulin in vitro, and the higher the absolute value of the electret surface potential, the better the effect of the insulin to the insulin. The distribution of insulin in the cuticle was not significantly different from that of the control group, but the distribution of insulin in the skin of the stratum corneum increased significantly. (3) -1500V electret combined with 3% azone, after 36 hours, the cumulative transdermal volume of insulin was 3% azone group, 1.4 times as much as 3% azone group and the control group, 2.12 times and 2.93 times.-1500V in the group of electret. The combination of polar body and azone with 3% azone has a good synergistic effect on insulin. (three) in vitro transdermal regulation of insulin through the skin of diabetic rats, a diabetic rat model was successfully constructed by injecting streptozotocin, and the insulin solution was examined by the insulin solution after the action of -1500V, -1000V, and -500V electret. The skin transdermal rule of diabetic rats showed that the skin permeation of diabetic rats was significantly higher than that of normal skin. The cumulative transmission of 36 hours was 3.2 times.36 hours after normal skin transmission, the insulin solution after the action of -1500V, -1000V, -500V electret to diabetic rats The cumulative permeation of the skin was 4.00,3.86,3.54 times of the normal skin transmission, the higher the absolute value of the surface potential, the more the increase. (four) the effect of electret on the microstructure of normal rats and diabetic skin was studied by using immunofluorescence technology and microstructural analysis techniques, and the effects of different polarity and surface potential electret were studied. The microstructure of normal and diabetic rats showed that 1) electret could thinner the cuticle of the skin of normal rats, increase the gap of the cells, enlarge the mouth of the hair follicle, and expand.2). After the electret acted on the skin of the diabetic rats, the cuticle and the dermis of the skin thickened slightly, and the insulin which had the repair effect of.3) the main transdermal path of the insulin after the action of the electret. The diameter is the structure of the hair follicle of the skin. (five) the effect of the electret on insulin hypoglycemic effect is similar to that of the insulin solution without electret acting on the electret body. The effect of insulin in the +1000V electret and the -1000V electret is better. The system of the electret insulin transdermal delivery system is stable. It is characterized by good quality, convenient use, good biological safety and no need for external instrument and equipment. The electret insulin transdermal delivery system is expected to provide new methods and means for patients with diabetes.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R587.1

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