Dermatology Department, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing, 10010, People's Republic of China.
School of Clinical Medicine, Beijing University of Chinese Medicine, Beijing, People's Republic of China.
Int J Nanomedicine. 2023 Jul 10;18:3781-3800. doi: 10.2147/IJN.S406649. eCollection 2023.
In this study, we aimed to report the biological characteristics of the first successful synthesis of gentiopicroside-loaded chitosan nanoparticles and to evaluate the therapeutic effects and preliminary mechanisms of gentiopicrin-loaded chitosan on psoriasis-like cell and mouse models.
Gentiopicroside-loaded chitosan nanoparticles (CHI-GEN) were prepared, and their biological characteristics were evaluated. HaCaT keratinocytes were stimulated with TNF-α to establish a psoriatic keratinocyte model. MTT assay and flow cytometry were used to measure cell viability and apoptosis, respectively. mRNA levels of K17, VEGF A, and IL-6 and IL-23A were detected using qRT-PCR. These tests were used to preliminarily assess the effects of CHI-GEN on keratinocyte proliferation and inflammation. Imiquimod was used to construct a psoriasis-like mice model. The severity of psoriasis was scored based on the psoriasis area severity index (PASI), H&E staining was used to observe the histological changes and the level of inflammation and cell proliferation of skin lesions was evaluated by measuring the mRNA levels of K17, IL-23A, and IL-17A using qRT-PCR.
The average particle size of CHI-GEN nanoparticles was approximately 100 nm, and the zeta potential was 2.69 ± 0.87 mV. The cumulative release was 67.2% in solutions of pH 5.5 at 24 h. GEN reduced TNF-α-induced excessive proliferation of HaCaT keratinocytes and downregulated mRNA levels of K17, VEGF A, and inflammatory cytokines IL-6 and IL-23A, which was more obvious in the CHI-GEN treatment group. Additionally, CHI-GEN significantly improved the severity of skin lesions in psoriasis-like mice and downregulated the mRNA expressions of IL-6, IL-23A, and IL-17A in mice skin lesions.
In conclusion, we successfully prepared gentiopicrin-chitosan nanoparticles. Our results show that these nanoparticles have anti-psoriasis activity, inhibits keratinocyte proliferation and improves symptoms in psoriasis model mice and can be used to develop an effective strategy for the treatment of psoriasis.
本研究旨在报道成功合成龙胆苦苷载壳聚糖纳米粒的生物学特性，并评价龙胆苦苷载壳聚糖对银屑病样细胞和小鼠模型的治疗作用及初步机制。
制备龙胆苦苷载壳聚糖纳米粒（CHI-GEN），并对其生物学特性进行评价。用 TNF-α刺激 HaCaT 角质形成细胞建立银屑病角质形成细胞模型。采用 MTT 法和流式细胞术分别检测细胞活力和凋亡。采用 qRT-PCR 检测 K17、VEGF A 和 IL-6、IL-23A 的 mRNA 水平。这些测试用于初步评估 CHI-GEN 对角质形成细胞增殖和炎症的影响。用咪喹莫特构建银屑病样小鼠模型。根据银屑病面积严重指数（PASI）对银屑病严重程度进行评分，H&E 染色观察组织学变化，采用 qRT-PCR 检测 K17、IL-23A 和 IL-17A 的 mRNA 水平评估皮肤病变的炎症和细胞增殖水平。
CHI-GEN 纳米粒的平均粒径约为 100nm，Zeta 电位为 2.69±0.87mV。在 pH5.5 的溶液中，24 h 时累积释放率为 67.2%。GEN 降低了 TNF-α诱导的 HaCaT 角质形成细胞过度增殖，并下调了 K17、VEGF A 和炎症细胞因子 IL-6、IL-23A 的 mRNA 水平，CHI-GEN 治疗组更为明显。此外，CHI-GEN 显著改善了银屑病样小鼠皮肤病变的严重程度，并下调了小鼠皮肤病变中 IL-6、IL-23A 和 IL-17A 的 mRNA 表达。
综上所述，我们成功制备了龙胆苦苷-壳聚糖纳米粒。我们的结果表明，这些纳米粒具有抗银屑病活性，抑制角质形成细胞增殖，并改善银屑病模型小鼠的症状，可用于开发治疗银屑病的有效策略。
