目的 制备绿原酸-黄芩苷共载纳米粒鼻腔原位凝胶（chlorogenic acid-baicalin co-loaded nanoparticles in situ nasal gel，CA&B-NPs/ISNG），初步考察其对上呼吸道黏膜免疫功能低下（upper airway immunity dysfunction，UAID）小鼠的影响。方法 以包封率和胶凝温度为评价指标，制备出CA&B-NPs/ISNG，表征其形态、粒径分布、ζ电位、体外释药量及凝胶溶蚀量；采用牛蛙上颚离体法考察CA&B-NPs/ISNG对纤毛的毒性；建立UAID小鼠模型，测定小鼠唾液的分泌型免疫球蛋白A（secretory immunoglobulin A，SIgA）含量和溶菌酶活性，考察CA&B-NPs/ISNG增强鼻黏膜免疫的作用。结果 优选处方：取绿原酸和黄芩苷各11.0 mg，加入10 mL聚乙二醇（PEG）400溶解成CGA-BC溶液，磁力搅拌下缓慢滴入到25 mL 2.72 mg/mL壳聚糖溶液中，调节pH至5.0，然后磁力搅拌下缓慢滴加10 mL 1.00 mg/mL三聚磷酸钠（tripolyphosphate，TPP）溶液，得CA&B-NPs混悬液；往CA&B-NPs混悬液中加入体系18.5%泊洛沙姆407（Poloxamer 407，P407），溶胀分散均匀后，加入体系1%泊洛沙姆188（Poloxamer 188，P188），待完全溶胀分散均匀，调节pH至5.5，得CA&B-NPs/ISNG。纳米粒形态规则，平均粒径（536.10±16.24）nm，粒子分散系数0.310±0.053，ζ电位（24.39±0.27）mV，绿原酸和黄芩苷包封率分别为（44.67±0.43）%和（55.25±0.61）%；胶凝温度为（32.03±0.31）℃。CA&B-NPs/ISNG表现出持续缓慢释药和无明显的纤毛毒性，且能显著提高SIgA的含量和溶菌酶的活性（P＜0.05）。结论 制备的CA&B-NPs/ISNG均一稳定，包封率高，适用鼻用制剂要求，可提高UAID小鼠的鼻黏膜免疫功能。

Objective To prepare for chlorogenic acid and baicalin co-loaded nanoparticles in situ nasal gel (CA&B-NPs/ISNG) and to investigate its effect on upper airway immunity dysfunction (UAID) mice. Methods To use encapsulation rate and gelation temperature as evaluation index, CA&B-NPs/ISNG was prepared. Then it was characterized by morphology, particle size distribution, ζpotential, drug release in vitro and gel corrosion amount. The toxicity of CA&B-NPs/ISNG to cilia was investigated by using toad palate mucosa in vitro. The UAID mice was established and the enhancement effect of CA&B-NPs/ISNG on nasal mucosal immunity was detected by measuring the SIgA content and lysozyme activity in saliva of mice. Results The preferred prescription is to take 11.0 mg each of chlorogenic acid and baicalin, 10 mL PEG400 was added to dissolve into chlorogenic acid- baicalin solution and slowly dropped into 25 mL 2.72 mg/mL chitosan solution with magnetic stirring. Adjusted the pH to 5.0. Then 10 mL 1.00 mg/mL TPP solution was slowly added dropwise under magnetic stirring to obtain CA&B-NPs suspension; CA&B-NPs was added to system 18.5% P407, swollen and dispersed well, then added system 1% P188, completely swollen and dispersed well, and the pH was adjusted to 5.5 to obtain CA&B-NPs/ISNG. Nanoparticles showed regular morphology with particle size (536.10 ±16.24) nm, ζ potential (24.39 ±0.27) mV, particle dispersion coefficient 0.310 ±0.053, chlorogenic acid and baicalin encapsulation rates of (44.67 ±0.43)% and (55.25 ±0.61)%, respectively. The gelling temperature was (32.03 ±0.31)℃. CA&B-NPs/ISNG showed sustained slow drug release and no obvious ciliary toxicity, and significantly increased SIgA content and lysozyme activity (P < 0.05). Conclusion The prepared CA&B-NPs/ISNG was stable and had high encapsulation rate. It was suitable for nasal preparation requirement and can improve nasal mucosal immune function in UAID mice.

R283.6

全国中医药创新骨干人才培养对象项目［国中医药人教函（2019）128号］