五谷虫通过抑制免疫应激-补体活化缓解咪喹莫特诱导的小鼠银屑病样皮肤损伤

doi:10.12122/j.issn.1673-4254.2024.11.08

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (11): 2121-2130.doi: 10.12122/j.issn.1673-4254.2024.11.08

• • 上一篇

五谷虫通过抑制免疫应激-补体活化缓解咪喹莫特诱导的小鼠银屑病样皮肤损伤

姚宏¹^,²(), 刘珂娣²(), 刘澄曌², 李伟红²^,³, 代旗², 赵石², 丁子恒¹^,², 王鹤霏¹^,², 葛晓静⁴, 卫培峰¹^,⁵, 段佳林⁶(), 奚苗苗²^,⁵()

^1.陕西中医药大学药学院，陕西咸阳 712046
^2.西安天工生物医药研究所有限公司，陕西西安 710032
^3.西北大学生命科学学院，陕西西安 710069
^4.中国人民解放军联勤保障部队第九八八医院采购管理科，河南郑州 450042
^5.陕西中医药大学第二附属医院国家药物临床试验机构，陕西咸阳 712021
^6.西北工业大学医学研究院，陕西西安 710072

收稿日期:2024-06-14 出版日期:2024-11-20 发布日期:2024-11-29
通讯作者: 段佳林,奚苗苗 E-mail:yao520hong1314@163.com;pepperyfat@163.com;dogson1989@163.com;miaomiaoxi2014@163.com
作者简介:姚宏，在读硕士研究生，E-mail: yao520hong1314@163.com
刘珂娣，硕士，E-mail：pepperyfat@163.com；
第一联系人：姚宏、刘珂娣共同为第一作者
基金资助:
上海市2022年度“科技创新行动计划”生物医药科技支撑专项(22S21902600)

Maggot alleviates imiquimod-induced psoriasis-like skin lesions in mice by inhibiting immune stress and complement activation

Hong YAO¹^,²(), Kedi LIU²(), Chengzhao LIU², Weihong LI²^,³, Qi DAI², Shi ZHAO², Ziheng DING¹^,², Hefei WANG¹^,², Xiaojing GE⁴, Peifeng WEI¹^,⁵, Jialin DUAN⁶(), Miaomiao XI²^,⁵()

^1.School of Pharmacy, Shaanxi University of Chinese Medicine, Xianyang 712046, China
^2.Xi'an TANK Medicinal Biology Institute, Xi'an 710032, China
^3.College of Life Sciences, Northwestern University, Xi'an 710069, China
^4.Procurement Management Department, 988 Hospital of PLA Joint Logistics Support Force, Zhengzhou 450042, China
^5.National Institute of Drug Clinical Trials, Second Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712021, China
^6.Institute of Medicine, Northwestern Polytechnical University, Xi'an 710072, China

Received:2024-06-14 Online:2024-11-20 Published:2024-11-29
Contact: Jialin DUAN, Miaomiao XI E-mail:yao520hong1314@163.com;pepperyfat@163.com;dogson1989@163.com;miaomiaoxi2014@163.com

摘要/Abstract

摘要：

目的研究五谷虫通过免疫应激-补体活化途径治疗C57BL/6银屑病样小鼠的药效学及作用机制。方法取36只SPF级雄性C57BL/6小鼠，随机分为正常组、模型组、五谷虫（1.25%，2.5%，5%剂量）组、本维莫德（1%）组，6只/组，咪喹莫特乳膏涂抹造模。MPASI评分检测小鼠皮损的严重程度；游标卡尺检测小鼠耳廓肿胀厚度；HE染色检测小鼠背部皮肤及右耳组织病理学变化；观察小鼠抓挠行为学；称重计算小鼠脾脏指数；甲苯胺蓝染色检测小鼠背部皮肤组织肥大细胞数量；ELISA法检测免疫应激（IgG、IgM）、补体活化（CH50、C1s、C3、C3a、C5、C5a）、炎症反应（IL-23、IL-17A、TNF-α）相关因子水平。结果与正常组相比，模型组小鼠MPASI评分、耳廓肿胀厚度均升高（P<0.001）；背部皮肤及右耳组织病理损伤加重；抓挠行为次数，脾脏指数，肥大细胞数量，血清IgG、C1s、C3a、C5a、IL-23、IL-17A、TNF-α，组织C1s、C3、C3a、C5、C5a水平均升高（P<0.05），血清CH50、C3、C5水平降低（P<0.001），血清IgM水平有升高趋势，但差异无统计学意义。与模型组相比，五谷虫低、中、高剂量组小鼠MPASI评分、耳廓肿胀厚度降低（P<0.05）；背部皮肤及右耳组织病理损伤减弱；抓挠行为次数、脾脏指数、肥大细胞数量降低（P<0.01），五谷虫高剂量组血清IgG、C1s、C3a、C5a、IL-23、IL-17A、TNF-α，组织C1s、C3、C3a、C5、C5a水平降低（P<0.05），血清CH50、C3、C5水平升高（P<0.01）。五谷虫疗效呈现剂量-效应依赖关系。结论五谷虫通过调控免疫应激-补体活化途径发挥治疗银屑病作用。

关键词: 五谷虫, 银屑病, 免疫应激, 补体活化, 炎症反应

Abstract:

Objective To explore the therapeutic mechanism of maggot for psoriasis-like lesions in mice from the perspective of immune stress and complement activation regulation. Methods Thirty-six male C57BL/6 mice were randomly divided into control group, model group, maggot (1.25%, 2.5%, and 5%) groups, and Benvitimod (1%) group. Psoriasis-like lesions were induced by application of imiquimod cream, and the severity of skin lesions was assessed using the modified Psoriasis Area and Severity Index (MPASI) score. Auricular swelling of the mice was observed, and histopathological changes of the skin lesions were examined with HE staining. Scratching behavior of the mice was observed and the spleen index was calculated. Toluidine blue staining was used to detect mast cells in the skin lesions, and serum levels of IgG, IgM, the complements CH50, C1s, C3, C3a, C5 and C5a, and the inflammatory factors IL-23, IL-17A and TNF-α were determined with ELISA. Results In mice with imiquimod-induced psoriasis-like skin lesions, treatment with the maggot at the 3 doses significantly decreased MPASI score, alleviated auricular swelling and pathologies in the skin lesions, reduced scratching behaviors, spleen index, and the number of mast cells in the lesions. Treatment with high-dose maggot significantly lowered serum levels of IgG, C1s, C3a, C5a, IL-23, IL-17A and TNF-α and the levels of C1s, C3, C3a, C5 and C5a in the lesion tissue, and increased serum levels of CH50, C3, and C5. The therapeutic effect of maggot showed a dose-effect dependence. Conclusion Maggot can alleviate psoriasis-like skin lesions in mice by inhibiting immune stress and complement activation.

Key words: maggot, psoriasis, immune stress, complement activation, inflammatory response

姚宏, 刘珂娣, 刘澄曌, 李伟红, 代旗, 赵石, 丁子恒, 王鹤霏, 葛晓静, 卫培峰, 段佳林, 奚苗苗. 五谷虫通过抑制免疫应激-补体活化缓解咪喹莫特诱导的小鼠银屑病样皮肤损伤[J]. 南方医科大学学报, 2024, 44(11): 2121-2130.

Hong YAO, Kedi LIU, Chengzhao LIU, Weihong LI, Qi DAI, Shi ZHAO, Ziheng DING, Hefei WANG, Xiaojing GE, Peifeng WEI, Jialin DUAN, Miaomiao XI. Maggot alleviates imiquimod-induced psoriasis-like skin lesions in mice by inhibiting immune stress and complement activation[J]. Journal of Southern Medical University, 2024, 44(11): 2121-2130.

图/表 10

表1 小鼠皮损面积及严重程度指数评分表

Tab.1 Psoriasis area and severity index in mice (MPASI)

Score	Severity	Erythema (E)	Infitration (I)	Desquamation (D)	Area of psoriatic involvement (%)
0	None	No erythema	No infitration	No scales	0
1	Slight	Slight erythema	The injured skin is slightly raised.	Some of the injured skin is covered with fine scales.	<10
2	Moderate	Moderate erythema	The injured skin is moderately raised, and the edge of the plaque is round or sloping.	Most of the injured skin is completely or incompletely covered with flaky scales.	10-29
3	Marked	Striking erythema	The injured skin is markedly raised and thickened.	Almost all of the injured skin is covered with thick and layered scales.	30-49
4	Very marked	Exceptionally striking erythema	The injured skin is very markedly raised and thickened.	All of the injured skin is covered with very thick and layered scales.	50-69
5	-	-	-	-	70-89
6	-	-	-	-	90-100

图1 五谷虫对PSO小鼠MPASI评分的影响

Fig.1 Effect of maggot MPASI score in psoriasis (PSO) mice. A: Skin lesions on day 1 and day 9 in the groups. B: Maggot significantly decreased MPASI score in PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; ###P<0.001 vs Model group.

图2 五谷虫对PSO小鼠耳廓肿胀厚度的影响

Fig.2 Effect of maggot on thickness of swelling auricle in PSO mice. A: Skin lesions in the right (shown on the left) and left ear (on the right) in mice. B: Maggot significantly decreased the auricle swelling thickness in PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; #P<0.05, ###P<0.001 vs Model group.

图3 五谷虫对PSO小鼠组织病理学变化的影响

Fig.3 Effect of maggot on histopathological changes in PSO mice. A: Maggot significantly improved pathological changes of back skin lesions in PSO mice (Scale bar=50 μm). B: Maggot significantly improved pathological changes in right ear lesions in PSO mice (Scale bar=100 μm).

图4 五谷虫对PSO小鼠抓挠行为学的影响

Fig.4 Maggot significantly decreased the number of scratching of PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; ##P<0.01, ###P<0.001 vs Model group.

图5 五谷虫对PSO小鼠脾脏指数的影响

Fig.5 Effect of maggot on spleen index in PSO mice. A: Comparison of mice spleen. B: Maggot significantly decreased the spleen index in PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; ###P<0.001 vs Model group.

图6 五谷虫对PSO小鼠组织肥大细胞数量的影响

Fig.6 Effect of maggot on the number of mast cells in PSO mice. A: Toluidine blue staining of dorsal skin lesions in mice (Scale bar=50 μm). B: Maggot significantly decreased the number of mast cells in PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; ##P<0.01, ###P<0.001 vs Model group.

图7 五谷虫对PSO小鼠免疫应激相关因子水平的影响

Fig.7 Effect of maggot on serum levels of IgG (A) and IgM (B) in PSO mice (Mean±SD, n=6). ***P<0.001 vs Control group; ##P<0.01 vs Model group.

图8 五谷虫对PSO小鼠补体活化相关因子水平的影响

Fig.8 Effect of maggot on levels of complements in PSO mice （Mean±SD, n=6）. A-F： Serum levels of CH50, C3, C5, C1s, C3a and C5a. G-K： C1s, C3, C3a, C5 and C5a contents in the skin tissue. *P<0.05, ***P<0.001vs Control group; #P<0.05, ##P<0.01, ###P<0.001 vs Model group.

图9 五谷虫对PSO小鼠炎症反应相关因子水平的影响

Fig.9 Effect of maggot on serum levels IL-23 (A), IL-17A (B) and TNF-α (C) in PSO mice (Mean±SD, n=6). **P<0.01, ***P<0.001 vs Control group; #P<0.05, ###P<0.001 vs Model group.

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五谷虫通过抑制免疫应激-补体活化缓解咪喹莫特诱导的小鼠银屑病样皮肤损伤

Maggot alleviates imiquimod-induced psoriasis-like skin lesions in mice by inhibiting immune stress and complement activation

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