脂质体鬼臼毒素在大鼠皮肤靶向性分布的初步实验研究
|
摘要
尖锐湿疣的发病率逐年增加，现已占我国性传播疾病的第二位。迄今为止，该病尚无有效的抗病毒治疗手段，并且部分尖锐湿疣可转化成鳞癌或宫颈癌，已发现90％的宫颈癌或宫颈上皮间变有HPV感染的证据。因此，尖锐湿疣已成为威胁人群健康的严重问题。治疗尖锐湿疣的方法很多，去除显而易见的赘生物相当容易，但无论那种方法，其复发率都较高。另外，HPV潜伏感染目前临床上尚无有效的诊断方法，而且是复发的主要原因。尖锐湿疣的发病、传播及蔓延难以预防和控制，治疗后的原位复发或它处新发往往在所难免。因此，需要对现在使用的药物加以改进。靶向性药物载体的研究是当前具有重大理论和应用价值的研究领域，而脂质体作为药物载体的研究是其中一个热点。用特定的脂质体包裹鬼臼毒素，使其向被HPV感染的表皮细胞浓集并缓慢释放药物，会在一定程度上改善鬼臼毒素的药物动力学和药效学特点，成为治疗HPV潜伏感染、防止CA复发和相关肿瘤发生的一种更有效的新方法。我们对此做了初步研究，旨在探讨脂质体鬼臼毒素在皮肤各层的分布规律。
本课题研究的目的：
1、探明脂质体鬼臼毒素在动物皮肤中的动态分布规律及其量化和成像方法。
2、推测脂质体鬼臼毒素外用的系统吸收性。
3、评估脂质体鬼臼毒素外用对大鼠肝肾功能的影响。
材料和方法：
1、按前期研究筛选出的合适配方制备脂质体鬼臼毒素制剂，用鬼臼毒素酊剂做对照，并按照随机对照的原则涂抹于实验大鼠的背部皮肤上。将大鼠皮肤制成切片后，用激光共聚焦显微成像仪观察鬼臼毒素荧光的分布情况，比较两种制剂鬼臼毒素在皮肤各层分布规律及其差异。
2、脂质体鬼臼毒素制剂和鬼臼毒素酊剂按照随机对照的原则涂抹于实验大鼠的背部皮肤上。用荧光分光光度法测量两种制剂不同
时间大鼠血中鬼臼毒素的浓度，将所得的结果进行统计分析，比
较两者的差异。
3、相同配方制备0．5％的脂质体鬼臼毒素制剂，用0．5％的鬼臼毒素
配剂做对照，并按照随机对照的原则涂抹于实验大鼠的背部皮肤
上，每大涂药2次，连续3天。涂药结束后4天取血测量肝肾功
能指标。
结果与分析：
l、应用激光共聚焦显微成像仪扫描和定量分析，表皮和真皮内鬼臼
毒素荧光值AUC，脂质体制剂分别是鬼臼毒素叮剂的1．5～2．3倍；
对照组在涂药2小时后，表皮和真皮的荧光量即达到峰值（分别
为1585．52／卜＊和2005石6／卜二‘），在4小时后荧光量迅速降低：
而观察组在4小时前，表皮和真皮中的荧光量较低，6小时后逐
渐增高，在1二刁时达峰值（分别为750．28／ 11＊2和10＊＊8／卜mz）。
2、对照组血中鬼臼毒素AUC是观察组的2．3倍。观察组用药8 ’J’
时后鬼臼毒素血药浓度达峰值，而对照组用药后2小时即达峰
值。观察组鬼臼毒素的峰值浓度较低，与对照组相比有显著性差
异（P＜0刀01）。
3、脂质体鬼臼毒素制剂外用后测定大鼠的肝肾功能指标与正常对
照组及配剂剂组相比均无显著性差异（P＞0．05）。
结论：
1、脂质体制剂能使鬼臼毒素在皮肤中缓慢释放，并持续较长时间。
2、脂质体鬼臼毒素混悬液外用，全身吸收明显低于外用鬼臼毒素叮
剂，表明脂质体鬼臼毒素外用后能降低全身毒副作用。
3、0．5舶质体鬼臼毒素混悬液外用，对大鼠肝肾功能没有影响，提
示其外用是安全可靠的。
The incidence rates of condyloma accuminatum(CA) is increasing annually. It is the second of sexually transmitted diseases (STDs) in our country now. There are no effective anti-virus therapeutic methods so far;and parts of CA can transform into squamous cell carcinoma or cervix cancer. The infection of HPV is discovered in 90% of carcinoma or anaplasia of cervix. Therefore,the CA has become the serious problem of threatening healthy of people. Many methods can treat warts and it is very easy to remove the obvious warts,however,any method has high recurrence rate. In addition,the latent infection of HPV that is main reason of recurrence has not effective method to diagnose clinically now. Therefore,the incidence,dissemination and spread of CA are difficult to prevent and control. It can hardly be avoided of recur in situation or in other places after therapy. Therefore,some progresses have to be made to treat the latent part. The study of targeting-drug vehicles is the important theory and applied worth at p
resent. As drug-delivery vehicles liposome is one of hot spots .Use of specific liposome encapsulated podophyllotoxin can accumulate and slowly release in epidermis cells infected of HPV,will improve ameliorate the drug's pharmacokinetic aspects and pharmacodynamic aspects. It may be a new method to suppress the latent infection of HPV or prevent recurrence of CA and related tumor. We are to make primary research to study distribution,regular pattern of liposome podophyllotoxin in skin. The purpose of this study:
1. To study distribution regular pattern of liposome podophyllotoxin in animal skin
2. Investigate the change of serum concentrations of podophyllotoxin after topical applying liposome-podophyllotoxin suspension on rat skin.
3. To evaluate the effect to hepatic and renal functions after liposome-podophyllotoxin topical application.
Materials and methods:
1. Liposome-podophyllotoxin was made ideal formulations according to early study methods,and control group was taken with podophyllotoxin alcohol solution. Two drugs were applied on rat skin under the principle of random and comparison. After a regulated time the skin was samples were taken and made into optical sections. Confocal laser scanning microscopy was used to detect the fluorescence,comparing their different and distribution regular pattern in skin.
2. Two kind of drugs,Liposome-podophyllotoxin and podophyllotoxin alcohol solution,were applied on rat skin under the principle of random and comparison. The serum concentration of podophyllotoxin was detected by spectrofluorometry. The results were discussed.
3. Liposome-podophyllotoxin was made ideal formulations according to early study methods,and control group was taken with podophyllotoxin alcohol solution. Two drugs were applied on rat skin under the principle of random and comparison. The drugs were applied on the rats skin twice a day for three days. To detect hepatic and renal functions after four days.
Results:
1. Confocal laser scanning microscopy shows that the AUC of fluorescent in epidermal and dermal test group is 1.5~2.3-fold higher than that of control. Unit area fluorescent amount of epidermal and dermal was highest after 2h in control group(l 585.527 u m2 and 2005.66 7u m2),and quick reduced after 4h;But unit area fluorescent amount of epidermal and dermal was rather low before 4h in observing group,and gradually increased after 6h,and was highest after 12h (750.287 u m2 and 1073.087 um2) .
2. The AUC of podophyllotoxin control group is 2.3-fold higher than that of the test. After 8h topical application,the serum concentration of podophyllotoxin was the highest in test group,while it reached in 2h as far as control group was concerned. The highest serum concentrations of podophyllotoxin in test group were significantly lower than those in control group (P<0.001).
3. There were no differences between test group and control group and
space group in hepatic and renal functions (P>0.05) Conclusion:
1. Podophyllotoxin of lipos
本课题研究的目的：
1、探明脂质体鬼臼毒素在动物皮肤中的动态分布规律及其量化和成像方法。
2、推测脂质体鬼臼毒素外用的系统吸收性。
3、评估脂质体鬼臼毒素外用对大鼠肝肾功能的影响。
材料和方法：
1、按前期研究筛选出的合适配方制备脂质体鬼臼毒素制剂，用鬼臼毒素酊剂做对照，并按照随机对照的原则涂抹于实验大鼠的背部皮肤上。将大鼠皮肤制成切片后，用激光共聚焦显微成像仪观察鬼臼毒素荧光的分布情况，比较两种制剂鬼臼毒素在皮肤各层分布规律及其差异。
2、脂质体鬼臼毒素制剂和鬼臼毒素酊剂按照随机对照的原则涂抹于实验大鼠的背部皮肤上。用荧光分光光度法测量两种制剂不同
时间大鼠血中鬼臼毒素的浓度，将所得的结果进行统计分析，比
较两者的差异。
3、相同配方制备0．5％的脂质体鬼臼毒素制剂，用0．5％的鬼臼毒素
配剂做对照，并按照随机对照的原则涂抹于实验大鼠的背部皮肤
上，每大涂药2次，连续3天。涂药结束后4天取血测量肝肾功
能指标。
结果与分析：
l、应用激光共聚焦显微成像仪扫描和定量分析，表皮和真皮内鬼臼
毒素荧光值AUC，脂质体制剂分别是鬼臼毒素叮剂的1．5～2．3倍；
对照组在涂药2小时后，表皮和真皮的荧光量即达到峰值（分别
为1585．52／卜＊和2005石6／卜二‘），在4小时后荧光量迅速降低：
而观察组在4小时前，表皮和真皮中的荧光量较低，6小时后逐
渐增高，在1二刁时达峰值（分别为750．28／ 11＊2和10＊＊8／卜mz）。
2、对照组血中鬼臼毒素AUC是观察组的2．3倍。观察组用药8 ’J’
时后鬼臼毒素血药浓度达峰值，而对照组用药后2小时即达峰
值。观察组鬼臼毒素的峰值浓度较低，与对照组相比有显著性差
异（P＜0刀01）。
3、脂质体鬼臼毒素制剂外用后测定大鼠的肝肾功能指标与正常对
照组及配剂剂组相比均无显著性差异（P＞0．05）。
结论：
1、脂质体制剂能使鬼臼毒素在皮肤中缓慢释放，并持续较长时间。
2、脂质体鬼臼毒素混悬液外用，全身吸收明显低于外用鬼臼毒素叮
剂，表明脂质体鬼臼毒素外用后能降低全身毒副作用。
3、0．5舶质体鬼臼毒素混悬液外用，对大鼠肝肾功能没有影响，提
示其外用是安全可靠的。
The incidence rates of condyloma accuminatum(CA) is increasing annually. It is the second of sexually transmitted diseases (STDs) in our country now. There are no effective anti-virus therapeutic methods so far;and parts of CA can transform into squamous cell carcinoma or cervix cancer. The infection of HPV is discovered in 90% of carcinoma or anaplasia of cervix. Therefore,the CA has become the serious problem of threatening healthy of people. Many methods can treat warts and it is very easy to remove the obvious warts,however,any method has high recurrence rate. In addition,the latent infection of HPV that is main reason of recurrence has not effective method to diagnose clinically now. Therefore,the incidence,dissemination and spread of CA are difficult to prevent and control. It can hardly be avoided of recur in situation or in other places after therapy. Therefore,some progresses have to be made to treat the latent part. The study of targeting-drug vehicles is the important theory and applied worth at p
resent. As drug-delivery vehicles liposome is one of hot spots .Use of specific liposome encapsulated podophyllotoxin can accumulate and slowly release in epidermis cells infected of HPV,will improve ameliorate the drug's pharmacokinetic aspects and pharmacodynamic aspects. It may be a new method to suppress the latent infection of HPV or prevent recurrence of CA and related tumor. We are to make primary research to study distribution,regular pattern of liposome podophyllotoxin in skin. The purpose of this study:
1. To study distribution regular pattern of liposome podophyllotoxin in animal skin
2. Investigate the change of serum concentrations of podophyllotoxin after topical applying liposome-podophyllotoxin suspension on rat skin.
3. To evaluate the effect to hepatic and renal functions after liposome-podophyllotoxin topical application.
Materials and methods:
1. Liposome-podophyllotoxin was made ideal formulations according to early study methods,and control group was taken with podophyllotoxin alcohol solution. Two drugs were applied on rat skin under the principle of random and comparison. After a regulated time the skin was samples were taken and made into optical sections. Confocal laser scanning microscopy was used to detect the fluorescence,comparing their different and distribution regular pattern in skin.
2. Two kind of drugs,Liposome-podophyllotoxin and podophyllotoxin alcohol solution,were applied on rat skin under the principle of random and comparison. The serum concentration of podophyllotoxin was detected by spectrofluorometry. The results were discussed.
3. Liposome-podophyllotoxin was made ideal formulations according to early study methods,and control group was taken with podophyllotoxin alcohol solution. Two drugs were applied on rat skin under the principle of random and comparison. The drugs were applied on the rats skin twice a day for three days. To detect hepatic and renal functions after four days.
Results:
1. Confocal laser scanning microscopy shows that the AUC of fluorescent in epidermal and dermal test group is 1.5~2.3-fold higher than that of control. Unit area fluorescent amount of epidermal and dermal was highest after 2h in control group(l 585.527 u m2 and 2005.66 7u m2),and quick reduced after 4h;But unit area fluorescent amount of epidermal and dermal was rather low before 4h in observing group,and gradually increased after 6h,and was highest after 12h (750.287 u m2 and 1073.087 um2) .
2. The AUC of podophyllotoxin control group is 2.3-fold higher than that of the test. After 8h topical application,the serum concentration of podophyllotoxin was the highest in test group,while it reached in 2h as far as control group was concerned. The highest serum concentrations of podophyllotoxin in test group were significantly lower than those in control group (P<0.001).
3. There were no differences between test group and control group and
space group in hepatic and renal functions (P>0.05) Conclusion:
1. Podophyllotoxin of lipos
引文
[1] Werness BA, The role of the HPV omotroteins in transformation and carcinogenic progression. Lippincott, 1991, 1: 2～18.
[2] 周华，杨帆，熊礼宽，等．尖锐湿疣人乳头瘤病毒潜伏感染的研究．中华皮肤科杂志．1995；28(3)：168～169．
[3] 孔繁荣，朱学骏．人乳头瘤病毒(HPV)携带者在STD门诊患者中的检出率．北京医科大学学报，1995；27(2)：156～159．
[4] Strand A, wilander E, Zehbe I, et al. High risk HPV persists after treatment of genital papillomavirus infection but not after treatment of cervical intraepithelial neoplasia[J]. Obstet Gynecol Scand Acta. 1997, 76(2): 140～144.
[5] Beutner KL, Ferenczy A. Therapeutic approaches to genital warts. Am J Med. 1997: 102(5A): 28～37.
[6] 朱文元．经足叶草脂、鬼臼毒素和5-Fu处理后的CA皮损中HPV-DNA检测．江苏医药，1999；25(3)：205．
[7] Kinghorn GR, Mcmillan A, Mulcahy F, et al. An open, comparative study of the efficacy of 0.5% podophyllotoxin lotion and 25%podophyllion solution in the treatment of condylomata acuminata in males and females[J]. Int J STDAIDS, 1993; 4: 194～199.
[8] Petersen CS, Weisamann K, Kaempherol Q. An argument against the use ofpodophyllin? [J]. Genitourin Med, 1995; 71(1): 92～93.
[9] De Villiers EM. Laboratory diagnosis in the investigation of human papillomaverus infection. Genitourinary medicine, 1992; 68: 50～54.
[10] 孔繁荣，朱学骏，瞿文学．尖锐湿疣人乳头瘤病毒感染的分型研究．中华皮肤科杂志，1994；27(3)：157～159．
[11] Bangham AD, Standish MM, Watkins JC, et al. Diffusion of univalentions across the lamellac of swollen phospholipids. J Mol Biol, 1965; 13(1): 238～252.
[12] Micheal M, Vijeyalakshmi G. Liposome: a selective drug delivery system for the topical route of administration: gel dosage form. J Pharm Pharmacoh 1982; 34(7): 473.
[13] Micheal M, Vijeyalakshmi G. Liposome: a selective drug delivery system for the topical route of administration lotion dosage form. Life Sci, 1980; 26: 1473.
[14] Mayer LD, Bally MB, Cullis PR, et al. Uptake of adfiamycin into large unilamellar vesicles in response to a pH gradient. Biochim Biophys Area, 1986; 857(1): 123～126.
[15] Haran G, Cohen R, Bar LK, et al. Transmembrane ammonium sulfate gradients in liposome produce efficient and stable entrapment of amphipathic weak bases. Biochim Biophys Atca, 1993; 1151(2): 201～215.
[16] Pavelic Z, Skalko BN, Jalsenjaki. Liposomes containing drugs for tretment of vaginal infections. Eur J Pharm Sci, 1999; 8(4): 345～351.
[17] Lasch J, Bouwstra J. Ineraction of external lipids(lipidvesicles) with the skin. J Liposome Res, 1995; 5(3): 543～569.
[18] Touitou E, Junginger HE, et al. liposomes as carriers for topical and transdermal delivery. J Pharmaceutical Res, 1994; 83(9): 1189～1203.
[19] Short S, Rubas W, et al. Transport of biologically active interferon-gamma across human skin in vitro. Pharmaceutical Res, 1995; 12(8): 1140～1146.
[20] Mezei M. Administration of drags with multiphase liposomal delivery system[P]. Patent, 1990; US 4897269.
[21] 马守栋，李国锋，陈志良等．脂质体鬼臼毒素的制备及质量研究．中国药业，1997；(5)：12～13
[22] 雷国华，鲍永耀，朴英杰等．共聚焦显微镜及其参数选择．中国医学物理学杂志，1997；14(1)：46～48．
[23] Kiyoshi K, Malgoriats S, Howard L. Percutaneous absorption: a single-layermodel. J Phrem Sci, 1993; 82(5): 450.
[24] Braun-Falco O, Korting HC, Maibach HI, et al. Biodisposition of liposome-encapsulated active ingredients applied on the skin. Liposome Dermatics, 1992: 206～214.
[25] 瞿光喜，邹立家，张天民．脂质体经皮给药的研究进展．中国药学杂志，1997；32(6): 329～332．
[26] Masini V, Bonte F, Meybeck A, et al. Cutaneous bioavailability in hairless rats of retinoic acid in liposome or gel[J]. J Pharm Sci, 1993; 82(1): 17～21.
[27] 张灵芝．脂质体制备及其在生物医学中的应用．第1版．北京：北京医科大学、中国协和医科大学联合出版社．1998年，第101页．
[28] Plessis JD, Weiner N, Muller PG. The influence of in vivo treatment of skin with liposomes on the topical absorption of a hydrophilic and a hydrophobic drug in vitro[J]. Int J Pharm, 1994: 103(1): R1-R5.
[29] Lasch J, Wohlrab W. Liposome-bound cortisol: a new approach to cutaneous therapy. Biomed Biochim Acta, 1996; 45: 1295～1299.
[30] 李国锋，许重远，孟庆礼，等．鬼臼毒素具有荧光性．第一军医大学学报，1996；16(3)：268～269．
[31] El-Ridy M. S, Khalil RM, Free versus liposome-encapsulated lignocaine hydrochloride yopical applications[J]. Pharmazie, 1999; 54(9): 682-684.
[32] Sharma BB, Jain SK, Sagar SP. Topical liposome system bearing local anesthetic lignocaine: preparation and evaluation. J Microencapsulation, 1994; 11(3): 279.
[33] Syrjanen SM. Basic concepts and practical applications of recombinant DNA techniques in detection of human papillomavims(HPV) infection. APMIS, 1990; 82(2): 95～110.
[34] Coderch L, De-Dera M, Perec-Cullell N, et al. The effect of liposomes on skin barrier structure. Skin Pharmaco Applica Skin Physico, 1999: 12(5): 235-246.
[35] Lopez-Amaya C, Marangoni AG. Comparison of dynamic and integrated light-scattering techniques in the study of the interaction of Candida rugosa lipase with DPPC liposomes. Biophys-Chem, 1999; 80(2): 69-83.
[36] Bernard E, Dubois JL, Wepierre J, et al. Importance of sebaceous glands in cutaneous penetration of an antiandrogen: target effect of liposomes[J]. J ParmaSci, 1997; 86(5): 573-578.
[37] Greenberg MD, Rutledge LH, Reid R, et al. A double-blind, randomized trial of 0.5% podofilod and plocebo for the treatment of genital warts in women[J]. Obstet Gynecol, 1991; 77: 735～739.
[38] Waldrep JC, Gilbert BE, Knight CM, et al. Pulmonary delivery of beclomethasone liposome aerosol in volunteers: tolerance and safety. Chest, 1997; 111(2): 316-323.
[39] Wyde PPR, Gilbert BE, Mehta K. Aerosol delivery of liposomal all-trans-retinoic acid to the lungs. Cancer Chemother Pharmacol, 1999; 43: 277-283.
[40] 曾抗，李国锋，许重远，等，脂质体鬼臼毒素软膏治疗尖锐湿疣的双盲随机对照试验。第一军医大学学报，1998；18(3)：246
[41] Wang ZJ, He YY, Huang CG, et al. Pharmacokinetics, tissue distribution and photodynamic therapy efficacy of liposomal-deliverd hypocrellin A, a potential photosensitizer for tumor therapy. Photoche Photobio, 1999; 70(5). 773-780.
[42] Chang LW, Yang CM, Chen CF, et al. Experimental podophyllotoxin (bajiaolian) poisoning: Ⅱ. Effects on the liver, intestine, kidney, pancreas and testis. Biomed-Environ-Sci, 1992; 5(4): 293-302.
[43] Claesson U, Lassus A, Happonen H, et al. Topical treatment of venereal warts: a comparative open study of podophyllotoxin cream versus solution[J]. Int J STD AIDS, 1996; 7: 429～434.
[44] Tyring S, Edwards L, Cherry LK, et al. Safety and efficacy of 0.5% podofilox gel in the treatment of anogenital warts [J]. Arch Dermatol, 1998; 134(1): 33～38.
[45] Reynolds M, Fraser PA, Lacey CJN, Audits of the treatment of genital warts: closing the feedback loop[J]. Iht J STD AIDS, 1996; 7: 347-352.
[46] Stewart CF. Use of etoposide in patients with organ dysfunction: pharmacokinetic and pharmacodynamic considerations. Cancer Chemother Pharmacol, 1994; 34 Suppl: S76-83.
[47] Dhawan D, Balasubramanian S, Amonkar AJ, et al. Chemopreventive effect of 4'-demethyl epipodophyllotoxin on DMBA/TPA-induced mouse skin carcinogenesis. Carcinogenesis, 1999; 20(6): 997-1003.
[48] Estey E, Thall PF, Mehta K, et al. Alteration in tretinoin pharmacokinetics follo□ing administration of liposomal all-trans-retinoic acid. Blood, 1996, 87 (9): 3650-3654.
[49] 赵红，郑俊民．新型的皮肤给药系统——脂质体．沈阳药科大学学报，1997；14(2)：148～150．
[50] 范健，杨栋培，翁帼英．脂质体载药减少阿霉素心脏毒性的实验研究．中国药理学通报，1997；13(1)：66-69．
[2] 周华，杨帆，熊礼宽，等．尖锐湿疣人乳头瘤病毒潜伏感染的研究．中华皮肤科杂志．1995；28(3)：168～169．
[3] 孔繁荣，朱学骏．人乳头瘤病毒(HPV)携带者在STD门诊患者中的检出率．北京医科大学学报，1995；27(2)：156～159．
[4] Strand A, wilander E, Zehbe I, et al. High risk HPV persists after treatment of genital papillomavirus infection but not after treatment of cervical intraepithelial neoplasia[J]. Obstet Gynecol Scand Acta. 1997, 76(2): 140～144.
[5] Beutner KL, Ferenczy A. Therapeutic approaches to genital warts. Am J Med. 1997: 102(5A): 28～37.
[6] 朱文元．经足叶草脂、鬼臼毒素和5-Fu处理后的CA皮损中HPV-DNA检测．江苏医药，1999；25(3)：205．
[7] Kinghorn GR, Mcmillan A, Mulcahy F, et al. An open, comparative study of the efficacy of 0.5% podophyllotoxin lotion and 25%podophyllion solution in the treatment of condylomata acuminata in males and females[J]. Int J STDAIDS, 1993; 4: 194～199.
[8] Petersen CS, Weisamann K, Kaempherol Q. An argument against the use ofpodophyllin? [J]. Genitourin Med, 1995; 71(1): 92～93.
[9] De Villiers EM. Laboratory diagnosis in the investigation of human papillomaverus infection. Genitourinary medicine, 1992; 68: 50～54.
[10] 孔繁荣，朱学骏，瞿文学．尖锐湿疣人乳头瘤病毒感染的分型研究．中华皮肤科杂志，1994；27(3)：157～159．
[11] Bangham AD, Standish MM, Watkins JC, et al. Diffusion of univalentions across the lamellac of swollen phospholipids. J Mol Biol, 1965; 13(1): 238～252.
[12] Micheal M, Vijeyalakshmi G. Liposome: a selective drug delivery system for the topical route of administration: gel dosage form. J Pharm Pharmacoh 1982; 34(7): 473.
[13] Micheal M, Vijeyalakshmi G. Liposome: a selective drug delivery system for the topical route of administration lotion dosage form. Life Sci, 1980; 26: 1473.
[14] Mayer LD, Bally MB, Cullis PR, et al. Uptake of adfiamycin into large unilamellar vesicles in response to a pH gradient. Biochim Biophys Area, 1986; 857(1): 123～126.
[15] Haran G, Cohen R, Bar LK, et al. Transmembrane ammonium sulfate gradients in liposome produce efficient and stable entrapment of amphipathic weak bases. Biochim Biophys Atca, 1993; 1151(2): 201～215.
[16] Pavelic Z, Skalko BN, Jalsenjaki. Liposomes containing drugs for tretment of vaginal infections. Eur J Pharm Sci, 1999; 8(4): 345～351.
[17] Lasch J, Bouwstra J. Ineraction of external lipids(lipidvesicles) with the skin. J Liposome Res, 1995; 5(3): 543～569.
[18] Touitou E, Junginger HE, et al. liposomes as carriers for topical and transdermal delivery. J Pharmaceutical Res, 1994; 83(9): 1189～1203.
[19] Short S, Rubas W, et al. Transport of biologically active interferon-gamma across human skin in vitro. Pharmaceutical Res, 1995; 12(8): 1140～1146.
[20] Mezei M. Administration of drags with multiphase liposomal delivery system[P]. Patent, 1990; US 4897269.
[21] 马守栋，李国锋，陈志良等．脂质体鬼臼毒素的制备及质量研究．中国药业，1997；(5)：12～13
[22] 雷国华，鲍永耀，朴英杰等．共聚焦显微镜及其参数选择．中国医学物理学杂志，1997；14(1)：46～48．
[23] Kiyoshi K, Malgoriats S, Howard L. Percutaneous absorption: a single-layermodel. J Phrem Sci, 1993; 82(5): 450.
[24] Braun-Falco O, Korting HC, Maibach HI, et al. Biodisposition of liposome-encapsulated active ingredients applied on the skin. Liposome Dermatics, 1992: 206～214.
[25] 瞿光喜，邹立家，张天民．脂质体经皮给药的研究进展．中国药学杂志，1997；32(6): 329～332．
[26] Masini V, Bonte F, Meybeck A, et al. Cutaneous bioavailability in hairless rats of retinoic acid in liposome or gel[J]. J Pharm Sci, 1993; 82(1): 17～21.
[27] 张灵芝．脂质体制备及其在生物医学中的应用．第1版．北京：北京医科大学、中国协和医科大学联合出版社．1998年，第101页．
[28] Plessis JD, Weiner N, Muller PG. The influence of in vivo treatment of skin with liposomes on the topical absorption of a hydrophilic and a hydrophobic drug in vitro[J]. Int J Pharm, 1994: 103(1): R1-R5.
[29] Lasch J, Wohlrab W. Liposome-bound cortisol: a new approach to cutaneous therapy. Biomed Biochim Acta, 1996; 45: 1295～1299.
[30] 李国锋，许重远，孟庆礼，等．鬼臼毒素具有荧光性．第一军医大学学报，1996；16(3)：268～269．
[31] El-Ridy M. S, Khalil RM, Free versus liposome-encapsulated lignocaine hydrochloride yopical applications[J]. Pharmazie, 1999; 54(9): 682-684.
[32] Sharma BB, Jain SK, Sagar SP. Topical liposome system bearing local anesthetic lignocaine: preparation and evaluation. J Microencapsulation, 1994; 11(3): 279.
[33] Syrjanen SM. Basic concepts and practical applications of recombinant DNA techniques in detection of human papillomavims(HPV) infection. APMIS, 1990; 82(2): 95～110.
[34] Coderch L, De-Dera M, Perec-Cullell N, et al. The effect of liposomes on skin barrier structure. Skin Pharmaco Applica Skin Physico, 1999: 12(5): 235-246.
[35] Lopez-Amaya C, Marangoni AG. Comparison of dynamic and integrated light-scattering techniques in the study of the interaction of Candida rugosa lipase with DPPC liposomes. Biophys-Chem, 1999; 80(2): 69-83.
[36] Bernard E, Dubois JL, Wepierre J, et al. Importance of sebaceous glands in cutaneous penetration of an antiandrogen: target effect of liposomes[J]. J ParmaSci, 1997; 86(5): 573-578.
[37] Greenberg MD, Rutledge LH, Reid R, et al. A double-blind, randomized trial of 0.5% podofilod and plocebo for the treatment of genital warts in women[J]. Obstet Gynecol, 1991; 77: 735～739.
[38] Waldrep JC, Gilbert BE, Knight CM, et al. Pulmonary delivery of beclomethasone liposome aerosol in volunteers: tolerance and safety. Chest, 1997; 111(2): 316-323.
[39] Wyde PPR, Gilbert BE, Mehta K. Aerosol delivery of liposomal all-trans-retinoic acid to the lungs. Cancer Chemother Pharmacol, 1999; 43: 277-283.
[40] 曾抗，李国锋，许重远，等，脂质体鬼臼毒素软膏治疗尖锐湿疣的双盲随机对照试验。第一军医大学学报，1998；18(3)：246
[41] Wang ZJ, He YY, Huang CG, et al. Pharmacokinetics, tissue distribution and photodynamic therapy efficacy of liposomal-deliverd hypocrellin A, a potential photosensitizer for tumor therapy. Photoche Photobio, 1999; 70(5). 773-780.
[42] Chang LW, Yang CM, Chen CF, et al. Experimental podophyllotoxin (bajiaolian) poisoning: Ⅱ. Effects on the liver, intestine, kidney, pancreas and testis. Biomed-Environ-Sci, 1992; 5(4): 293-302.
[43] Claesson U, Lassus A, Happonen H, et al. Topical treatment of venereal warts: a comparative open study of podophyllotoxin cream versus solution[J]. Int J STD AIDS, 1996; 7: 429～434.
[44] Tyring S, Edwards L, Cherry LK, et al. Safety and efficacy of 0.5% podofilox gel in the treatment of anogenital warts [J]. Arch Dermatol, 1998; 134(1): 33～38.
[45] Reynolds M, Fraser PA, Lacey CJN, Audits of the treatment of genital warts: closing the feedback loop[J]. Iht J STD AIDS, 1996; 7: 347-352.
[46] Stewart CF. Use of etoposide in patients with organ dysfunction: pharmacokinetic and pharmacodynamic considerations. Cancer Chemother Pharmacol, 1994; 34 Suppl: S76-83.
[47] Dhawan D, Balasubramanian S, Amonkar AJ, et al. Chemopreventive effect of 4'-demethyl epipodophyllotoxin on DMBA/TPA-induced mouse skin carcinogenesis. Carcinogenesis, 1999; 20(6): 997-1003.
[48] Estey E, Thall PF, Mehta K, et al. Alteration in tretinoin pharmacokinetics follo□ing administration of liposomal all-trans-retinoic acid. Blood, 1996, 87 (9): 3650-3654.
[49] 赵红，郑俊民．新型的皮肤给药系统——脂质体．沈阳药科大学学报，1997；14(2)：148～150．
[50] 范健，杨栋培，翁帼英．脂质体载药减少阿霉素心脏毒性的实验研究．中国药理学通报，1997；13(1)：66-69．