Development of a new therapy for endometriosis using biodegrative drug delivery system

子宫内膜异位症的治疗方法是全身应用激素调节剂，如促性腺激素释放激素激动剂、口服避孕药和达那唑(DZ)。这些治疗方法可以改善症状，缩小病灶大小，但疗效有限，长期治疗往往会引发严重的不良反应。我们设计了一种新的药物释放系统，包括DZ负载的透明质酸(HA)，用于局部应用于子宫内膜异位症。制备了含1或10 mg/ml DZ的疏水改性HA水凝胶(DZ-HA凝胶)，无菌注射入大鼠子宫内膜异位症模型。观察包囊大小、囊壁和血液中DZ浓度、处理包囊的组织学和发情周期的变化。免疫组织化学和[~(3 H)]-胸腺嘧啶核苷掺入法检测子宫内膜上皮细胞的增殖活性。即使在雌激素…存在的情况下，10^&lt；-5&gt；MDZ也能抑制培养的子宫内膜上皮细胞的[~3H]-TdR参入。更多的ol(10^&lt；-8&gt；M)，表明DZ直接抑制。注射后4周，未加DZ的HA水凝胶组(n=8)、1 mg/mlDZ-HA凝胶组(n=8)、10 mg/mlDZ-HA凝胶组(n=8)、去卵巢大鼠(n=8)的包囊体积分别减少到72.2±10.9%、71.3±8.4%、89.2±8.8%和12.7±2.5%。组织学上，DZ-HA凝胶处理的包囊显示子宫内膜上皮显著萎缩。10 mg/mlDZ-HA凝胶组细胞凋亡率增加，增殖细胞数减少。10 mg/mlDZ-HA凝胶处理的包囊在注射后3周的DZ浓度为3000~13,300 ng/g组织，但血浆DZ浓度从未超过5.0 ng/ml，所有DZ-HA凝胶处理的大鼠(n=16)都维持了正常的发情周期。局部注射DZ-HA凝胶可在不干扰性周期的情况下使实验性子宫内膜异位症模型消退。这些结果表明，使用这种新型给药系统的DZ局部应用可能被证明是治疗子宫内膜异位症的有效方法。较少

Endometriosis has been treated using systemic administration of hormonal modulators such as GnRH agonists, oral contraceptives and danazol (DZ). These treatments improve symptoms and reduce lesion size, but effectiveness is limited and prolonged treatment often provokes serious adverse effects. We devised a novel drug delivery system comprising DZ-loaded hyaluronic acid (HA) for local application to endometriosis. Hydrogels of hydrophobically modified HA with DZ loaded at concentrations of 1 or 10 mg/ml (DZ-HA gel) were prepared and injected aseptically into a rat model of endometriosis. Changes in cyst size, DZ concentrations in cyst walls and blood, histology of treated cysts, and estrous cycles were examined. Proliferative activity of endometrial epithelial cells was evaluated immunohistologically and using [^3H]-thymidine incorporation assay. Treatment with 10^<-5> M DZ inhibited [^3H]-thymidine incorporation in cultured endometrial epithelial cells, even in the presence of estradi … More ol (10^<-8> M), suggesting direct inhibition by DZ. Size of treated cysts decreased to 72.2 ±10.9% for HA hydrogel without DZ (n=8), 71.3 ±8.4% for 1 mg/ml DZ-HA gel (n=8), 89.2 ±8.8% for 10 mg/ml DZ-HA gel (n=8), and 12.7 ±2.5% for ovariectomized rats (n=8) at 4 weeks after injection. Histologically, DZ-HA gel-treated cysts displayed marked atrophy of the endometrial epithelium. Increased numbers of apoptotic cells and decreased numbers of proliferative cells were noted with 10 mg/ml DZ-HA gel. DZ concentrations in cysts treated using 10 mg/ml DZ-HA gel were 3000〜13,300 ng/g tissue at 3 weeks after injection, but plasma levels of DZ never exceeded 5.0 ng/ml. Regular estrous cycles were maintained in all DZ-HA gel-treated rats (n=16). Local injection of DZ-HA gel achieved regression of an experimental model of endometriosis without disturbing the sexual cycle. These results suggest that local application of DZ using this novel drug delivery system may prove useful for treating endometriosis. Less