生肌玉红膏提取物对斑马鱼皮肤创面愈合的促进作用及其机制

doi:10.13481/j.1671-587X.20250508

吉林大学学报(医学版) ›› 2025, Vol. 51 ›› Issue (5): 1221-1229.doi: 10.13481/j.1671-587X.20250508

• 基础研究 • 上一篇

生肌玉红膏提取物对斑马鱼皮肤创面愈合的促进作用及其机制

滕欢欢¹,孙光^1,²,姜锐²,孙立伟²(),刘建增³()

^1.长春中医药大学药学院，吉林长春 130117
^2.长春中医药大学附属医院中医药研究中心，吉林长春 130021
^3.长春中医药大学东北亚中医药研究院中药所，吉林长春 130117

收稿日期:2024-11-08 接受日期:2024-12-19 出版日期:2025-09-28 发布日期:2025-11-05
通讯作者: 孙立伟,刘建增 E-mail:sunnylilwei@163.com;liujz_1988@163.com
作者简介:滕欢欢（1997-），女，黑龙江省绥化市人，在读硕士研究生，主要从事中药健康产品研究和开发方面的研究。
基金资助:
吉林省科技厅科技发展计划项目(YDZJ202201ZYTS283);吉林省中医药管理局中医药科技项目(2022123)

Promotive effect of Shengji Yuhong Ointment extract on skin wound healing of zebrafish and its mechanism

Huanhuan TENG¹,Guang SUN^1,²,Rui JIANG²,Liwei SUN²(),Jianzeng LIU³()

^1.School of Pharmacy，Changchun University of Chinese Medicine，Changchun 130117，China
^2.Research Center of Traditional Chinese Medicine，Affiliated Hospital，Changchun University of Chinese Medicine，Changchun 130021，China
^3.Institute of Traditional Chinese Medicine，Northeast Asia Institute of Traditional Chinese Medicine，Changchun 130117，China

Received:2024-11-08 Accepted:2024-12-19 Online:2025-09-28 Published:2025-11-05
Contact: Liwei SUN,Jianzeng LIU E-mail:sunnylilwei@163.com;liujz_1988@163.com

摘要/Abstract

摘要：

目的探究生肌玉红膏提取物（SYOE）对斑马鱼皮肤创面愈合的促进作用，并阐明其作用机制。方法取320条野生型AB系斑马鱼，于侧腹部切口建立创面模型后，随机分为对照组、低剂量SYOE组（置于含0.625 mg·L^-1 SYOE的水中饲养）、高剂量SYOE组（置于含1.250 mg·L^-1 SYOE的水中饲养）和尿囊素组（置于含1.000 mg·L^-1尿囊素的水中饲养），每组80条。分别于损伤后第7、14和21天拍照记录斑马鱼皮肤创面面积，计算创面愈合率。于不同时间点收集各组斑马鱼创面皮肤组织制作组织切片，采用HE染色观察第0、7、14和21天各组斑马鱼皮肤创面边缘宽度，天狼星红染色检测损伤后第2、4、6和8天各组斑马鱼皮肤创面组织中的胶原蛋白水平，酶联免疫吸附试验（ELISA）法检测损伤后第4天各组斑马鱼皮肤创面组织中Ⅰ型胶原蛋白（Col Ⅰ）和α-平滑肌肌动蛋白（α-SMA）水平，实时荧光定量PCR（RT-qPCR）法分析损伤后第4天各组斑马鱼皮肤创面组织中Col Ⅰ编码基因col1a1a、col1a1b、col1a2和α-SMA以及转化生长因子β（TGF-β）/Smad信号通路关键因子tgfb1a、smad2和smad3a mRNA表达水平。结果创面愈合评估，与对照组比较，损伤后第7、14和21天低剂量SYOE组、高剂量SYOE组及尿囊素组斑马鱼皮肤创面愈合率明显升高（P<0.05或P<0.01），其中高剂量SYOE组在第21天时斑马鱼皮肤创面愈合率达84%。HE染色，与对照组比较，损伤后第14和21天，低剂量SYOE组、高剂量SYOE组及尿囊素组斑马鱼皮肤创面边缘宽度明显减小（P<0.05或P<0.01）。天狼星红染色，损伤后第6和8天，与对照组比较，低剂量SYOE组、高剂量SYOE组和尿囊素组斑马鱼皮肤创面组织中胶原蛋白水平明显升高（P<0.01）。ELISA法检测，损伤后第4天，与对照组比较，低剂量SYOE组、高剂量SYOE组和尿囊素组斑马鱼皮肤创面组织中Col Ⅰ及α-SMA水平明显升高（P<0.05或P<0.01）。RT-qPCR法检测，损伤后第4天，与对照组比较，低剂量SYOE组斑马鱼皮肤创面组织中col1a1b、col1a2、α-SMA、tgfb1a和smad2 mRNA表达水平明显升高（P<0.05或P<0.01），高剂量SYOE组和尿囊素组斑马鱼皮肤创面组织中col1a1a、col1a1b、col1a2、α-SMA、tgfb1a、smad2及smad3a mRNA表达水平明显升高（P<0.01）。结论 SYOE能够增加斑马鱼皮肤创面的胶原蛋白沉积，促进皮肤创面愈合，并提高TGF-β/Smad信号通路相关基因的表达水平。

关键词: 生肌玉红膏提取物, 创面愈合, 斑马鱼, 胶原蛋白沉积, 转化生长因子β, Smad

Abstract:

Objective To discuss the promotive effect of Shengji Yuhong Ointment extract （SYOE） on skin wound healing of zebrafish， and to clarify its mechanism. ? Methods ?A total of 320 wild-type AB strain zebrafish were used to establish a wound model by making an incision on the lateral abdomen. The zebrafish were randomly divided into control group， low dose of SYOE group （raised in water containing 0.625 mg·L^-1 SYOE）， high dose of SYOE group （raised in water containing 1.250 mg·L^-1 SYOE）， and allantoin group （raised in water containing 1.000 mg·L^-1 allantoin）， and there were 80 zebrafish in each group. The wound areas were recorded by photographs on days 7， 14， and 21 after injury， and the wound healing rates were calculated. The skin tissue samples from the wound sites of the zebrafish in various groups were collected at different time points to prepare histological sections. HE staining was used to observe the widths of skin wound edges of the zebrafish on days 0， 7， 14， and 21 in various groups； Sirius red staining was used to detect the collagen levels in the wound tissues of the zebrafish on days 2， 4， 6， and 8 after injury in various groups； enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of type Ⅰ collagen （Col Ⅰ） and α-smooth muscle actin （α-SMA） in the wound tissues on day 4 after injury； real-time quantitative PCR （RT-qPCR） was used to analyze the mRNA expression levels of Col Ⅰ-encoding genes （col1a1a， col1a1b， and col1a2） and α-SMA as wall as key factors of the transforming growth factor β （TGF-β）/Smad signaling pathway （tgfb1a， smad2， and smad3a） in the wound tissues of the zebrafish in various groups on day 4 after injury.?? Results The wound healing assessment results showed that compared with control group， the wound healing rates of the zebrafish in low dose of SYOE group， high dose of SYOE group， and allantoin group were significantly increased on days 7， 14， and 21 after injury （P<0.05 or P<0.01）； the wound healing rate in high dose of SYOE group achieved 84% on day 21. The HE staining results showed that compared with control group， the widths of skin wound edges of the zebrafish in low dose of SYOE group， high dose of SYOE group， and allantoin group were significantly decreased on days 14 and 21 after injury （P<0.05 or P<0.01）.The Sirius red staining results showed that compared with control group， the collagen levels in the skin wound tissue of the zebrafish in low dose of SYOE group， high dose of SYOE group， and allantoin group were significantly increased on days 6 and 8 after injury （P<0.01）. The ELISA results showed that on day 4 after injury， compared with control group， the levels of Col Ⅰ and α-SMA in the skin wound tissue of the zebrafish in low dose of SYOE group， high dose of SYOE group， and allantoin group were significantly increased （P<0.05 or P<0.01）. The RT-qPCR results showed that on day 4 after injury， compared with control group， the expression levels of col1a1b， col1a2， α-SMA， tgfb1a， and smad2 mRNA in the skin wound tissue of the zebrafish in low dose of SYOE group were significantly increased （P<0.05 or P<0.01）， while the expression levels of col1a1a， col1a1b， col1a2， α-SMA， tgfb1a， smad2， and smad3a mRNA in the skin wound tissue of the zebrafish in high dose of SYOE group and allantoin group were significantly increased （P<0.01）. Conclusion SYOE can increase the collagen deposition in skin wound of zebrafish， promote wound healing， and upregulate the expression of genes related to the TGF-β/Smad signaling pathway.

Key words: Shengji Yuhong Ointment extract, Wound healing, Zebrafish, Collagen deposition, Transforming growth factor β, Smad

中图分类号:

R285.5

滕欢欢,孙光,姜锐,孙立伟,刘建增. 生肌玉红膏提取物对斑马鱼皮肤创面愈合的促进作用及其机制[J]. 吉林大学学报(医学版), 2025, 51(5): 1221-1229.

Huanhuan TENG,Guang SUN,Rui JIANG,Liwei SUN,Jianzeng LIU. Promotive effect of Shengji Yuhong Ointment extract on skin wound healing of zebrafish and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1221-1229.

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Primer	Sequence（5'-3'）
tgfb1a F	GCACCGTGATACTCCATGACTTG
tgfb1a R	GATCTCTGACCTGATGTAGAATGTTCC
α-SMA F	GTGTTCCGCTGTTGCCTATGAG
α-SMA R	ACTGGTTACGCTGCTTGATGTTAC
smad2 F	GACGCAAGACCGCTGAAGAAC
smad2 R	TTCATTCGCTGTATGCTCTGCTAAG
smad3a F	GCCCACGCAGCAGCAAAC
smad3a R	ATTGTATCCTCCATCGGTTGTTCAC
col1a1a F	ACCGTAATGTGCGACGAAGT
col1a1a R	CCTGGAAATCGTCGTCTGGG
col1a1b F	TTTCTGCTAGGGTCTGGATTGG
col1a1b R	AGCTGCAGTAACTTCGTCGG
col1a2 F	GCTGGTCAAGATGGTCGCA
col1a2 R	CAGCCTCGCCAGAAGGTC
GAPDH F	CTTGCTGTAGTAAGTGTTTGCTTGC
GAPDH R	AACCTGGTGCTCCGTGTATCC

Group	Col Ⅰ[ρ_B/（μg·L^-1）]	α-SMA[ρ_B/（ng·L^-1）]
Control	16.06±2.39	14.95±0.12
Low dose of SYOE	19.05±0.19^*	18.26±0.29 ^**
High dose of SYOE	20.13±0.11^**	22.97±0.19 ^**
Allantoin	21.00±0.37^**	24.42±1.32 ^**

Group	col1a1a mRNA	col1a1b mRNA	col1a2 mRNA	α-SMA mRNA
Control	1.01±0.02	1.00±0.00	1.00±0.08	1.03±0.02
Low dose of SYOE	1.02±0.03	1.20±0.04 ^**	1.16±0.01 ^**	1.63±0.09 ^*
High dose of SYOE	1.37±0.04 ^**	1.34±0.02 ^**	1.25±0.04 ^**	2.51±0.05 ^**
Allantoin	1.46±0.04 ^**	1.39±0.05 ^**	1.31±0.05 ^**	2.80±0.34 ^**

Group	tgfb1a mRNA	smad2 mRNA	smad3a mRNA
Control	1.00±0.00	1.03±0.02	1.00±0.05
Low dose of SYOE	1.15±0.02 ^*	1.34±0.04 ^*	1.14±0.08
High dose of SYOE	1.27±0.05 ^**	1.48±0.01 ^**	1.36±0.09 ^**
Allantoin	1.35±0.07 ^**	2.18±0.15 ^**	1.58±0.05 ^**

生肌玉红膏提取物对斑马鱼皮肤创面愈合的促进作用及其机制

Promotive effect of Shengji Yuhong Ointment extract on skin wound healing of zebrafish and its mechanism

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