目的：制备一种以聚乳酸-羟基乙酸共聚物(PLGA)为载体，丝裂霉素C(MMC)搭载于PLGA的纳米药物MMC-ATS-@PLGA，并从体内实验角度分析该纳米药物对于青光眼术后抑制滤过泡瘢痕增殖的生物安全性和治疗效果。

方法：采用薄膜分散-水化超声法制备纳米药物MMC-ATS-@PLGA，并检测其物理和化学性质。通过角膜荧光染色和HE染色分析MMC-ATS-@PLGA对兔眼角膜的影响，并进一步收集兔眼泪膜破裂时间(BUT)、Schirmer试验、眼压等数据分析眼表生物安全性。通过裂隙灯观察并计算滤过泡大小和结膜HE染色分析组织形态学变化，并通过免疫组化和Elisa实验多角度基于蛋白组学对比分析氟米龙滴眼液(FML)、MMC和MMC-ATS-@PLGA纳米粒抑制青光眼术后滤过泡瘢痕效果。

结果：MMC-ATS-@PLGA平均粒径和电位为128.78±2.54 nm和36.49±4.25 mV，其包封率和载药率分别为(78.49±2.75)%和(30.86±1.84)%； 在33 ℃(眼表温度)条件下，MMC-ATS-@PLGA纳米粒在600 min时累计释放率可达(76.58±2.68)%。同时角膜荧光染色、HE、BUT、Schirmer和眼压结果显示，MMC-ATS-@PLGA对于兔眼表生物安全性良好。术后3 wk，结果显示MMC-ATS-@PLGA组滤过泡的面积明显大于FML组和MMC组，控制组滤过泡基本消失； 进一步提取兔眼滤过泡区域的结膜进行病理组织分析，MMC-ATS-@PLGA组胶原纤维形态相对规则，排列整齐，组织形态基本接近正常组。并且Elisa实验结果表明，与正常组相比，控制组中α-SMA、CTGF和Ⅲ型胶原纤维抗体的表达水平显著增高，予以FML、MMC和MMC-ATS-@PLGA治疗3 wk后，炎症因子表达逐渐下降，其中MMC-ATS-@PLGA组炎症因子表达下降最明显(P<0.05)。

结论：本研究成功合成一种针对青光眼滤过术后抑制滤过泡瘢痕增殖的纳米药物(MMC-ATS-@PLGA)，其具有稳定的物理化学性质、良好的生物相容性和更佳的抗炎效果，其通过抑制炎症因子α-SMA、CTGF和Ⅲ型胶原纤维的表达，阻止滤过泡区瘢痕化形成，从而提高青光眼滤过手术的成功率。

METHODS: The thin-film dispersion hydration ultrasonic method was used to prepare the MMC-ATS-@PLGA, and its physical and chemical properties were detected. The effect of MMC-ATS@PLGA on rabbit corneas was analysed through corneal fluorescence staining and HE staining, and tear film rupture time(BUT), Schirmer test and intraocular pressure data were collected to analyse ocular surface biosafety. A slit lamp was used to observe and calculate the filtration bubble size, and the tissue morphological changes were analysed by conjunctival HE staining. In addition, immunohistochemistry and Elisa were used to compare the anti-inflammatory effects of Flumiolone Eye Drops(FML), MMC, and MMC-ATS-@PLGA nanoparticles on inhibiting the formation of filtering bleb scarring after glaucoma surgery from multiple perspectives via comparative proteomic analysis.

RESULTS: The average particle size and zeta potential of MMC-ATS-@PLGA were 128.78±2.54 nm and 36.49±4.25 mV, respectively, with an encapsulation efficiency and a drug loading rate of(78.49±2.75)% and(30.86±1.84)%, respectively. At 33°C(the ocular surface temperature), the cumulative release rate of the MMC-ATS-@PLGA nanoparticles reached(76.58±2.68)% after 600 min. Moreover, corneal fluorescence staining, HE, BUT, Schirmer, and intraocular pressure results showed that MMC-ATS-@PLGA had good biocompatibility with the ocular surface of rabbits. At 3 wk after surgery, the area of filtering blebs in the MMC-ATS-@PLGA group was significantly larger than that in the FML group and MMC group, and the filtering blebs in the control group had basically disappeared. Pathological tissue analysis of the conjunctiva in the filtering blebs area of the eyes of the rabbits revealed that compared with that in the normal group, the morphology of the collagen fibres in the MMC-ATS-@PLGA group was relatively regular, the fibres were arranged neatly, and the tissue morphology was similar to that of the normal group. Immunohistochemistry and Elisa confirmed that compared with those in the normal group, the expression levels of α-SMA, CTGF, and type Ⅲ collagen fibre antibodies were significantly increased in the control group. After FML, MMC, or MMC-ATS-@PLGA treatment for 3 wk, the expression of inflammatory factors gradually decreased. Among the groups, the MMC-ATS-@PLGA group showed the most significant decrease(P<0.05).

CONCLUSION: This study successfully synthesized a nanomedicine(MMC-ATS-@PLGA)that inhibits scar proliferation after glaucoma filtration surgery. The drug had stable physicochemical properties, good biocompatibility, and better anti-inflammatory effects by inhibiting the expression of α-SMA, CTGF, and type Ⅲ collagen fibres, which can prevent the formation of scarring in the filtering blebs area, thereby improving the success rate of glaucoma filtering surgery.