鬼臼毒素纳米脂质载体的制备及用于西藏小型猪宫颈黏膜的安全性研究

发布时间：2018-06-12 12:48

本文选题：鬼臼毒素 + 纳米脂质载体　；参考：《南方医科大学》2017年硕士论文

【摘要】：研究背景鬼臼毒素(podophyllotoxin,POD)是从足叶草及八角莲的根茎中提取的一种具有显著抗病毒和抗肿瘤作用的天然活性木脂素。1990年WHO将鬼臼毒素酊剂(podophyllotoxin tincture,POD-T)外用作为治疗生殖器疣的一线治疗方案。然而,POD-T对生殖道黏膜刺激性大,使宫颈局部组织感染风险增加,且容易被吸收中毒,故不适合用于女性宫颈尖锐湿疣(condyloma acuminatum,CA)的治疗。20世纪以来,纳米脂质载体作为纳米制剂新剂型已广泛应用于临床研究,在生殖道黏膜给药中具有较好的应用前景。研究目的自制0.5%POD-NLC并检测其理化特性。建立西藏小型猪宫颈给药模型,以POD-酊剂为对照组,探讨其对小型猪宫颈黏膜的刺激性及系统毒性,进一步评价其安全性。材料与方法1、采用改良乳化蒸发-低温固化法,自制0.5%POD-NLC。考察0.5%POD-NLC外观、粒径、包封率及微观形态。2、选用不同剂量0.5%POD-NLC作为实验组,0.5%POD-T作阳性对照,观察宫颈黏膜的肉眼及病理改变;LUMINEX检测各实验组中宫颈分泌物IL-1β、IL-6、IL-8 的浓度。3、选用0.5%POD-NLC及不同浓度POD-T(0.5%,1%,2%),分别于小型猪宫颈给药2h,4h,6h,8h,10h后检测宫颈分泌物、宫颈黏膜组织POD浓度,评估0.5%POD-NLC的体内代谢规律。4、将不同剂量药物经小型猪宫颈黏膜给药,分别检测给药后2h、4h、6h、8h、10h血药浓度,分析0.5%POD-NLC经小型猪宫颈黏膜给药后血液中POD浓度变化,评估系统安全性。5、经宫颈黏膜给药后予小型猪前腔静脉取血,采用全自动生化仪测定各类生化指标;全自动血细胞分析仪测定各类血细胞数及血红蛋白含量,评估POD-NLC经小型猪宫颈黏膜给药后其血液生理生化指标改变。6、采用SPSS20.0(PASW)进行数据统计、描述与分析,数据以X±S表示。P0.05代表组间比较具有统计学差异。结果与分析1、成功制备0.5%POD-NLC,外观为质地均匀半透明胶状乳剂。马尔文型粒径分析仪检测POD-NLC粒径为178.5±20nm,包封率为(75.7±0.44)%,电镜扫描分析仪显示POD-NLC呈规则圆形及类圆形,分布均匀一致。2、小型猪宫颈黏膜刺激性评分结果显示,POD-T组总分值显著高于同等剂量POD-NLC组,差异具有统计学意义(p=0.001)。POD-NLC大剂量组总分值显著高于POD-NLC小剂量组,而生理盐水(NS)组总分值较POD-NLC小剂量组略低,无显著差异(p=0.092)。小型猪宫颈分泌物各因子浓度检测结果提示,POD-NLC小剂量组IL-1β、IL-6、IL-8浓度显著低于POD-NLC大剂量组(p0.01),而与NS组相比无明显差异(p0.05)。POD-T组各因子浓度较POD-NLC大剂量组升高,差异具有统计学意义(p0.05)。3、在相同给药时间内,0.5%POD-NLC组宫颈黏液中POD浓度均高于0.5%POD-T组(p0.05)。0.5%POD-T组宫颈黏膜组织中药物浓度最高峰值出现在给药后4h,随后快速下降。给药1Oh内,0.5%POD-NLC组黏膜中药物浓度呈缓慢持续升高,未见明显峰值;至给药10h,0.5%POD-NLC组黏膜组织中药物浓度开始高于0.5%POD-T组。4、宫颈给药1ml时,0.5%POD-NLC组未能测及POD血药浓度;0.5%POD-T组血药浓度最高峰值出现在给药后4h,并于给药后10h内呈低浓度持续出现。宫颈给药2ml时,0.5%POD-T组血药浓度显著高于0.5%POD-NLC组,差异具有显著意义(P0.05),0.5%POD-NLC组血药浓度变化平缓,未见明显峰值,给药10h后血药浓度未能测及。5、小型猪经POD-NLC、POD-T宫颈黏膜给药后,ALT、AST平均测定值均较给药前略高,但差异无统计学意义(p0.05)。各剂量组给药后白细胞测定值均较给药略高,但差异无统计学意义(p0.05)。0.5%POD-NLC(2ml)经小型猪宫颈黏膜给药后,未见明显系统毒性。结论本研究证实自制POD-NLC具有粒径小、稳定性好、包封率高等特点。POD-NLC经小型猪宫颈给药后局部刺激性小,同时可长时间滞留于宫颈分泌物中,并于宫颈黏膜组织中持续缓慢释药;常规治疗剂量POD-NLC对宫颈黏膜刺激性小,且经黏膜吸收量少,具有较好的系统安全性。
[Abstract]:Background podophyllotoxin (POD) is a natural active lignan, extracted from the rhizomes of podocyte and octagonal lotus, is a first-line treatment for the treatment of genital warts by using podophyllotoxin tincture (POD-T) as a first-line treatment for genital warts by.1990 WHO, which has a significant antiviral and anti-tumor effect. The mucosal irritation of the mucous membrane is large, the risk of local tissue infection of the cervix increases, and it is easily absorbed and poisoned, so it is not suitable for the treatment of condyloma acuminatum (CA) in the female cervix of the cervix for.20 century. The nano lipid carrier has been widely used in clinical research as a new dosage form of nanoscale preparation. The purpose of this study was to study the physical and chemical properties of the self-made 0.5%POD-NLC and to establish the physical and chemical properties of the cervix in Tibet miniature pigs. With the POD- tincture as the control group, the irritation and systemic toxicity of the cervix on the cervix mucosa of the miniature pig was discussed. The material and method 1, the modified emulsification evaporation low temperature curing method and the self-made 0.5%POD-NLC. were used. The 0.5%POD-NLC appearance, particle size, encapsulation rate and Micromorphology.2 were investigated, and different doses of 0.5%POD-NLC were selected as the experimental group, and 0.5%POD-T was used as the positive control to observe the naked eye and pathological changes of the cervical mucosa; LUMINEX was used to detect the IL-1 beta, IL-6, IL-8 concentration.3 in the experimental groups, and 0.5%POD-NLC and POD-T (0.5%, 1%, 2%), respectively, respectively. 2H, 4h, 6h, 8h, 10h were given to the cervix of a small pig to detect the cervical secretion and the concentration of POD in the cervical mucosa of the cervix, and to evaluate the metabolism of 0.5%POD-NLC in the body,.4. The drug was given at different doses of the cervix of the cervix of the small pig. The concentration of 2h, 4h, 6h, 8h, and the concentration of the blood were measured. Degree changes, evaluate the safety of the system.5. After the cervix mucosa was administered to the small pig's anterior vena cava, all kinds of biochemical indexes were measured by an automatic biochemical analyzer. The total number of blood cells and hemoglobin content were measured by the automatic blood cell analyzer. The blood physiological and biochemical indexes of POD-NLC after the administration of the cervix mucous membrane of the small pigs were changed.6. SPSS20.0 (PASW) data statistics, description and analysis, the data is X + S to indicate that.P0.05 represents a statistical difference between groups. Results and analysis 1, a successful preparation of 0.5%POD-NLC, the appearance of a homogeneous semitransparent gelatinous emulsion. The Malvin particle size analyzer is 178.5 + 20nm, the encapsulation rate is (75.7 + 0.44)%, SEM scanning The analyzer showed that POD-NLC was round and round, and the distribution of.2 was uniform and uniform. The results of cervical mucosa irritation of small pigs showed that the total score of POD-T group was significantly higher than that of the same dose POD-NLC group. The difference was statistically significant (p=0.001) the total score of.POD-NLC large dose group was significantly higher than that of the POD-NLC small dose group, while the total score of the normal saline (NS) group was significantly higher than that of the POD-NLC group. Compared with the POD-NLC small dose group, there was no significant difference (p=0.092). The results showed that the concentration of IL-1 beta, IL-6 and IL-8 in the POD-NLC small dose group was significantly lower than the POD-NLC large dose group (P0.01), but there was no significant difference between the NS group and the NS group (P0.05).POD-T group (P0.05).POD-T group. Statistical significance (P0.05).3, in the same time of administration, the concentration of POD in the cervical mucus in group 0.5%POD-NLC was higher than that in group 0.5%POD-T (P0.05) the highest concentration of drug concentration in the cervical mucous tissue of group.0.5%POD-T was found after the administration of 4h, and then decreased rapidly. In the drug 1Oh, the drug concentration in the mucous membrane of the 0.5%POD-NLC group increased slowly and continuously, and no obvious peak was found. When the drug was given 10h, the drug concentration in the 0.5%POD-NLC group began to be higher than that of the 0.5%POD-T group.4, and when the cervix was given 1ml, the 0.5%POD-NLC group failed to measure the concentration of POD; the highest peak value of the serum concentration in 0.5%POD-T group appeared at 4h after the administration, and the low concentration of the 10h was low in 10h after administration. The concentration of the blood drug in the 0.5%POD-T group was significant when the cervix was administered 2ml. Higher than 0.5%POD-NLC group, the difference was significant (P0.05), the blood drug concentration in group 0.5%POD-NLC was slow and no obvious peak, and the concentration of blood drug after 10h was not measured and.5 was not measured. After POD-NLC, POD-T cervix mucosa was given, ALT and AST mean values were slightly higher than those before administration, but the difference was not statistically significant (P0.05). The value of leucocyte was slightly higher than that of the drug, but the difference was not statistically significant (P0.05).0.5%POD-NLC (2ml) had no obvious systemic toxicity after the small pig cervix mucous membrane. Conclusion this study confirmed that the self-made POD-NLC has small size, good stability and high encapsulation efficiency, and.POD-NLC has a small local irritation after the administration of the cervix of a small pig, and it can be long at the same time. Time was delayed in cervix secretion and sustained slow release in cervical mucosa. The routine treatment dose of POD-NLC was less irritant to cervical mucosa and less mucosal absorption, and had better system safety.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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