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

Abstract

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

CLC Number:

R285.5

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.

Figures/Tables 10

Tab.1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Tab.2

Tab.3

Tab.4

References 32

[1]	WU H F， LI Z T， TANG J Q， et al. The in vitro and in vivo anti-melanoma effects of hydroxyapatite nanoparticles： influences of material factors［J］. Int J Nanomedicine， 2019， 14： 1177-1191.
[2]	PERSINAL-MEDINA M， LLAMES S， CHACÓN M， et al. Polymerizable skin hydrogel for full thickness wound healing［J］. Int J Mol Sci， 2022， 23（9）： 4837.
[3]	张婧. 巨噬细胞在皮肤伤口愈合过程中的调控作用及其机制研究［D］. 西安：西北大学， 2019.
[4]	YUAN M， LIU K， JIANG T， et al. GelMA/PEGDA microneedles patch loaded with HUVECs-derived exosomes and Tazarotene promote diabetic wound healing［J］. J Nanobiotechnology， 2022， 20（1）： 147.
[5]	CAO C W， XIAO Z C， TONG H Q， et al. Oral intake of chicken bone collagen peptides anti-skin aging in mice by regulating collagen degradation and synthesis， inhibiting inflammation and activating lysosomes［J］. Nutrients， 2022， 14（8）： 1622.
[6]	YU Q， SHEN Y X， XIAO F Q， et al. Yuhong ointment ameliorates inflammatory responses and wound healing in scalded mice［J］. J Ethnopharmacol， 2023， 306： 116118.
[7]	陈若岚，薛慧琴，丁劲，等. 艾灸联合生肌玉红膏对压疮大鼠皮肤损伤修复的作用机制［J］. 陕西中医， 2024， 45（8）： 1042-1047.
[8]	YANG Y W， ZHOU Y W， GE M L. The effect of externally applied traditional Chinese medicine in diabetic foot： a systematic review and meta-analysis of 34 RCTs［J］. Foot， 2023， 56： 102045.
[9]	唐丽利，冯文哲，石鹏，等. 加味生肌玉红膏促进湿热证肛瘘术后创面愈合的实验研究［J］. 时珍国医国药， 2023， 34（8）： 1798-1802.
[10]	刘恒，李爱茹，高岩，等. 基于微环境生态观察生肌玉红膏对犬咬伤创面愈合的促进作用［J］. 现代中医临床， 2023， 30（6）： 7-11.
[11]	GREENSPAN L J， AMEYAW K K， CASTRANOVA D， et al. Live imaging of cutaneous wound healing after rotary tool injury in zebrafish［J］. J Invest Dermatol， 2024， 144（4）： 888-897.e6.
[12]	WILKINSON H N， IVESON S， CATHERALL P， et al. A novel silver bioactive glass elicits antimicrobial efficacy against Pseudomonas aeruginosa and Staphylococcus aureus in an ex vivo skin wound biofilm model［J］. Front Microbiol， 2018， 9： 1450.
[13]	EDIRISINGHE S L， RAJAPAKSHA D C， NIKAPITIYA C， et al. Spirulina maxima derived marine pectin promotes the in vitro and in vivo regeneration and wound healing in zebrafish［J］. Fish Shellfish Immunol， 2020， 107： 414-425.
[14]	RICHARDSON R， SLANCHEV K， KRAUS C， et al. Adult zebrafish as a model system for cutaneous wound-healing research［J］. J Invest Dermatol， 2013， 133（6）： 1655-1665.
[15]	XIE H J， BAI Q， KONG F K， et al. Allantoin-functionalized silk fibroin/sodium alginate transparent scaffold for cutaneous wound healing［J］. Int J Biol Macromol， 2022， 207： 859-872.
[16]	ZHU H C， AO H T， FU Y L， et al. Optimizing alginate dressings with allantoin and chemical modifiers to promote wound healing［J］. Int J Biol Macromol， 2024， 275（Pt 1）： 133524.
[17]	CHEN Y X， ZHANG X R， LIU Z H， et al. Obstruction of the formation of granulation tissue leads to delayed wound healing after scald burn injury in mice［J］. Burns Trauma， 2021， 9： tkab004.
[18]	HE S Q， WALIMBE T， CHEN H Y， et al. Bioactive extracellular matrix scaffolds engineered with proangiogenic proteoglycan mimetics and loaded with endothelial progenitor cells promote neovascularization and diabetic wound healing［J］. Bioact Mater， 2021， 10： 460-473.
[19]	ZHOU W C， ZI L， CEN Y， et al. Copper sulfide nanoparticles-incorporated hyaluronic acid injectable hydrogel with enhanced angiogenesis to promote wound healing［J］. Front Bioeng Biotechnol， 2020， 8： 417.
[20]	ZHAO R L， JIN X Y， LI A， et al. Precise diabetic wound therapy： PLS nanospheres eliminate senescent cells via DPP4 targeting and PARP1 activation［J］. Adv Sci， 2022， 9（1）： 2104128.
[21]	YANG X Y， ZHI X Y， SONG Z L， et al. Flesh quality of hybrid grouper （Epinephelus fuscoguttatus ♀× Epinephelus lanceolatus ♂） fed with hydrolyzed porcine mucosa-supplemented low fishmeal diet［J］. Anim Nutr， 2022， 8（1）： 114-124.
[22]	XIE Y J， QIAO K， YUE L N， et al. A self-crosslinking， double-functional group modified bacterial cellulose gel used for antibacterial and healing of infected wound［J］. Bioact Mater， 2022， 17： 248-260.
[23]	CUI H S， JOO S Y， CHO Y S， et al. Effect of combining low temperature plasma， negative pressure wound therapy， and bone marrow mesenchymal stem cells on an acute skin wound healing mouse model［J］. Int J Mol Sci， 2020， 21（10）： 3675.
[24]	NAGASAKI K， NAKASHIMA A， TAMURA R， et al. Mesenchymal stem cells cultured in serum-free medium ameliorate experimental peritoneal fibrosis［J］. Stem Cell Res Ther， 2021， 12（1）： 203.
[25]	WU S Y， CHEN Y T， TSAI G Y， et al. Protective effect of low-molecular-weight fucoidan on radiation-induced fibrosis through TGF-β1/Smad pathway-mediated inhibition of collagen I accumulation［J］. Mar Drugs， 2020， 18（3）： 136.
[26]	邵世清，曲长萍，李好山，等. 脂联素干预对宫腔粘连大鼠子宫内膜组织中NLRP3、TGF-β1和Smad2表达的影响［J］. 郑州大学学报（医学版）， 2024， 59（6）： 777-782.
[27]	KIM H G， LIM Y S， HWANG S， et al. Di-（2-ethylhexyl） phthalate triggers proliferation， migration， stemness， and epithelial-mesenchymal transition in human endometrial and endometriotic epithelial cells via the transforming growth factor-β/Smad signaling pathway［J］. Int J Mol Sci， 2022， 23（7）： 3938.
[28]	PENG Y， WU S， TANG Q Y， et al. KGF-1 accelerates wound contraction through the TGF-β1/Smad signaling pathway in a double-paracrine manner［J］. J Biol Chem， 2019， 294（21）： 8361-8370.
[29]	HAU H T A， OGUNDELE O， HIBBERT A H， et al. Maternal Larp6 controls oocyte development， chorion formation and elevation［J］. Development， 2020， 147（4）： dev187385.
[30]	PALUNGWACHIRA P， TANCHAROEN S， PHRUKSANIYOM C， et al. Antioxidant and anti-inflammatory properties of anthocyanins extracted from Oryza sativa L. in primary dermal fibroblasts［J］. Oxid Med Cell Longev， 2019， 2019： 2089817.
[31]	LEE J J， NG S C， NI Y T， et al. Protective effects of galangin against H₂O₂/UVB-induced dermal fibroblast collagen degradation via hsa-microRNA-4535-mediated TGFβ/Smad signaling［J］. Aging， 2021， 13（23）： 25342-25364.
[32]	CHENG Y T， LI Y F， HUANG S Y， et al. Hybrid freeze-dried dressings composed of epidermal growth factor and recombinant human-like collagen enhance cutaneous wound healing in rats［J］. Front Bioeng Biotechnol， 2020， 8： 742.

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 ^**

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

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 32

Related Articles 15

Metrics

Comments

Recommended 0

[1]	Yingxin RUAN,Junya JIA,Zhanfei WU,Wenya SHANG,Pengyu ZHANG. Effect of NLRP3 inflammatome in renal interstitial fibrosis induced by unilateral ureteral obstruction in rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2024, 50(3): 587-595.
[2]	Meng QYU,Yuzhu JIANG,Rui HUANG,Zhenzhuo MA,Jichen XIA,Yuan ZHANG,Zhiheng DONG. Protective effect of ginsenoside Rh2 on kidney injury in rats with diabetic kidney lesions and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2023, 49(5): 1168-1173.
[3]	Yajing WANG,Baohui JIA,Fangfang HAN,Yingdong HAN,Lanlan ZHAO,Ying SHI,Hua ZHANG. Effect of human circular RNA_0114428 on lipopolysaccharide-induced injury of human renal tubular epithelial HK-2 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2023, 49(1): 101-109.
[4]	Junxiong ZHAO,Qian WU,Liangui NIE,Shengquan LIU,Zhentao JIANG,Jian CHEN,Ting XIAO,Jun YANG. Ameliorative effect of SO₂ on myocardial fibrosis in type 2 diabetes mellitus rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2023, 49(1): 8-14.
[5]	Meng QYU,Hong ZHENG,Yan LI,Boxue CHEN,Yuzhu JIANG,Shenggao WANG,Chunyan YU,Zhiheng DONG. Inhibitory effect of fermented red ginseng total saponins high glucose-induced renal tubular cell epithelial-mesenchymal transition and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(5): 1182-1189.
[6]	Yunfeng MA,Xiaofei HAN. Effects of salvianolate on osteoclast differentiation and bone resorption in osteoporotic rats by regulating SMAD2/FKBP1A/NF-κB axis [J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 111-121.
[7]	Zetai WANG,Dandan LOU,Yan PENG,Daoqi ZHU,Aiwu LI,Fengying GONG,Ying LYU,Qin FAN. Establishment of zebrafish xenograft model of nasopharyngeal carcinoma and inhibitory effect of curcumin on CNE-2 cells [J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 9-17.
[8]	Meng QU,Shiya WENG,Hong ZHENG,Yan LI,Runze GAO,Shenggao WANG,Chunyan YU,Boxue CHEN,Zhiheng DONG. Protective effect of fermented red ginseng total saponins on rat myocardial interstitial fibroblasts cultured with high glucose and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1201-1208.
[9]	Dan ZHANG,Ju HUI,Jiajuan GUO. Improvement effect of Jiedu Tongluo Decoction on myocardial fibrosis in rats through TGF-β1/Smad2/3 and ROCK pathways [J]. Journal of Jilin University(Medicine Edition), 2021, 47(4): 826-833.
[10]	Bo MA, Jiangang LI, Jun WANG, Junli HOU, Liang LI. Promotion effect of miR-106b on invasion and migration of colon cancer cells through targeting TGF-β/Smad pathway [J]. Journal of Jilin University(Medicine Edition), 2021, 47(3): 630-636.
[11]	Tianqi ZOU,Lin BAI,Zhengxu ZHANG,He LI,Chunmei WANG,Jinghui SUN,Jianguang CHEN,Xi CHEN,Chengyi ZHANG. Inhibitory effect of prunus tomentosa thunb total flavonoids on skin hypertrophic scar in mice and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 265-274.
[12]	Chunying ZHANG,Guangwei YIN,Mingda YOU,Hong CHEN,Yaojie HU,Yanbing LI,Chunyou CHEN. Inhibitory effects of siRNA targeting silencing TAK1 gene on proliferation and migration of thyroid cancer cells and p38 MAPK signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2021, 47(1): 110-117.
[13]	QIAO Liangwei, QU Qingshan, LI Ming. Effect of LncRNA-ATB on acute rejection of kidney transplanted rats by regulating miR-200c expression [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 955-962.
[14]	WEI Xujing, LI Lin, ZHANG Hongzhen, WANG Jing, XU Jing. Effects of LncRNA CCAT1 on proliferation,invasion and migration of endometrial cancer cells through TGF-β1/smad signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 1016-1022.
[15]	ZHAO Liping, HUANG Shubing, ZHANG Boping, ZHOU Zhilan, JIA Xuebing, SUN Mengfei, QIAO Chenmeng, CHEN Xue, SHEN Yanqin, CUI Chun. Inhibitory effect of Lactobacillus rhamnosus on intestinal inflammation after spinal cord injury in zebrafishes and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 680-686.