携载木犀草苷的透明质酸钠水凝胶宫腔灌注对大鼠宫腔粘连的疗效观察

doi:10.11958/20252034

摘要/Abstract

摘要：

目的探讨携载木犀草苷的透明质酸钠水凝胶宫腔灌注对宫腔粘连Sprague-Dawley（SD）大鼠模型的治疗作用。方法将50只SPF级SD大鼠随机分为空白对照组（A组）、宫腔粘连模型对照组（B组）、透明质酸钠水凝胶治疗组（C组）、木犀草苷溶液局部注射治疗组（D组）和携载木犀草苷的透明质酸钠水凝胶治疗组（E组），每组10只。除A组仅接受气体麻醉外，其余各组均采用95%乙醇注射法建立宫腔粘连模型。造模1周后，B—E组分别向子宫角内注射生理盐水、透明质酸钠水凝胶、10 g/L木犀草苷溶液以及携载10 g/L木犀草苷的透明质酸钠水凝胶。干预7 d后，采用Western blot检测子宫内膜组织中基质金属蛋白酶9（MMP-9）、转化生长因子β1（TGF-β1）、波形蛋白（Vimentin）、Ⅴ型胶原A2亚型（COL5A2）及Ⅰ型胶原A1亚型（COL1A1）蛋白的表达，酶联免疫吸附试验检测血清白细胞介素（IL）-6和IL-8水平；于妊娠2周后观察孕囊数量。结果与A组比较，B—E组TGF-β1、Vimentin、COL5A2、COL1A1、IL-6、IL-8表达水平升高，MMP-9蛋白表达水平降低，孕囊数量减少（P<0.05）。与B组相比，C—E组TGF-β1、Vimentin、COL5A2、COL1A1、IL-6、IL-8表达水平降低，MMP-9蛋白表达水平升高，孕囊数量增加（P<0.05），且E组上述指标变化较为显著。结论携载木犀草苷的透明质酸钠水凝胶宫腔灌注可有效减轻宫腔粘连，抑制炎症反应，提高孕囊数量，且其疗效优于单一治疗。

关键词: 组织黏连, 子宫疾病, 木犀草苷, 透明质酸, 水凝胶类

Abstract:

Objective To investigate the therapeutic efficacy of intrauterine perfusion with sodium hyaluronate hydrogel loaded with luteolin in a rat model of intrauterine adhesion (IUA). Methods Fifty specific pathogen free (SPF)-level SD rats were randomly assigned to five experimental groups: the normal control group (group A), the IUA model group (group B), the sodium hyaluronate hydrogel treatment IUA group (group C), the luteolin solution treatment IUA group (group D) and the sodium hyaluronate hydrogel loaded with luteolin treatment IUA group (group E), with 10 rats in each group. Except for the group A, which only received gas anesthesia, IUA model was established in all other groups using a 95% ethanol injection method. One week after modeling, rats in the groups B, C, D and E received intrauterine injection of normal saline, sodium hyaluronate hydrogel, 10 g/L luteoloside solution and sodium hyaluronate hydrogel loaded with 10 g/L luteoloside, respectively. Following a 7-day intervention period, protein expression levels of matrix metalloproteinase-9 (MMP-9), transforming growth factor-β1 (TGF-β1), Vimentin, collagen type Ⅴ alpha 2 chain (COL5A2) and collagen type I alpha 1 chain (COL1A1) in endometrial tissue were assessed by Western blot analysis. Serum concentrations of interleukin-6 (IL-6) and IL-8 were measured using enzyme-linked immunosorbent assays (ELISA). Two weeks post-intervention, the number of gestational sacs was recorded following pregnancy induction. Results Compared with the group A, the expression levels of TGF-β1, vimentin, COL5A2, COL1A1, IL-6 and IL-8 were increased, MMP-9 protein expression was decreased and the number of gestational sacs was reduced in the groups B, C, D and E (P < 0.05). Compared with the group B, expression levels of TGF-β1, vimentin, COL5A2, COL1A1, IL-6 and IL-8 decreased, MMP-9 protein expression increased and number of gestational sacs increased in the groups C, D and E (P < 0.05). These changes were more pronounced in the group E. Conclusion Intrauterine perfusion of sodium hyaluronate hydrogel loaded with luteoloside effectively alleviates intrauterine adhesions, suppresses inflammatory responses and enhances reproductive outcomes in the rat IUA model, demonstrating superior therapeutic efficacy compared to monotherapy approaches.

Key words: tissue adhesions, uterine diseases, luteoloside, hyaluronic acid, hydrogels

中图分类号:

R33，R711.32

图/表 3

参考文献 25

[1]	邢娉婷, 王爱丽, 陈彦辛. 补肾祛瘀法对中重度宫腔粘连的抗纤维化作用及经血中细胞外基质降解相关因子的影响[J]. 广州中医药大学学报, 2024, 41(1):96-104.
	XIN P T, WANG A L, CHEN Y X, et al. Anti-fibrosis action of therapy of tonifying kidney and removing stasis on moderate-to-severe intrauterine adhesions and its influence on extracellular matrix degradation related factors in menstrual blood[J]. Journal of Guangzhou University of Chinese Traditional Medicine, 2024, 41(1):96-104. doi:10.13359/j.cnki.gzxbtcm.2024.01.015.
[2]	姚欣怡, 俞凌. 宫腔粘连治疗的新进展[J]. 国际生殖健康/计划生育杂志, 2023, 42(5):431-436.
	YAO X Y, YU L. Advances of therapy for intrauterine adhesions[J]. J Int Reprod Health/Fam Plan, 2023, 42(5):431-436. doi:10.12280/gjszjk.20230224.
[3]	王丽, 张梦婷, 陈阳, 等. 人羊膜上皮干细胞联合富血小板血浆对宫腔粘连大鼠纤维化因子、炎症因子的作用[J]. 现代生物医学进展, 2023, 23(8):1430-1435.
	WANG L, ZHANG M T, CHEN Y, et al. Effect of human amniotic epithelial cells combined with platelet rich plasma on fibrotic factors and inflammatory factors in rats with intrauterine adhesions[J]. Progressin Modern Biomedicine, 2023, 23(8):1430-1435. doi:10.13241/j.cnki.pmb.2023.08.006.
[4]	王晓戈, 张靖笛, 李月, 等. 木犀草苷抑制SNHG14表达改善SW1353骨关节炎细胞模型的实验研究[J]. 中医药导报, 2023, 29(11):1-5.
	WANG X G, ZHANG J D, LI Y, et al. Inhibition of luteolin on SNHG14 expression to improve SW1353 osteoarthritis cell model[J]. Guid J Tradit Chin Med Pharm, 2023, 29(11):1-5. doi:10.13862/j.cn43-1446/r.2023.11.001.
[5]	秦璐. 木犀草苷对运动性疲劳大鼠的心肌氧化应激及抗氧化基因表达的影响[J]. 分子植物育种, 2023, 21(9):2900-2907.
	QIN L. Effects of cynaroside on myocardial oxidative stress and expression of antioxidant genes in exercise-induced fatigue rats[J]. Mol Plant Breed, 2023, 21(9):2900-2907. doi:10.13271/j.mpb.021.002900.
[6]	李男, 高关羽, 聂桐. 木犀草苷通过调控miR-211表达对缺氧缺糖诱导的大鼠皮质神经细胞损伤的影响[J]. 中国药师, 2021, 24(5):839-844.
	LI N, GAO G Y, NIE T. Effects of luteolin on oxygen-glucose deprivation induced cortical nerve cell damage in rats by regula-ting miR-211 expression[J]. China Pharmacist, 2021, 24(5):839-844. doi:10.3969/j.issn.1008-049X.2021.05.009.
[7]	XIONG F, LV X W. Luteolin reversed anxiety and depressive-like behavior via modulation of the NF-κB/NLRP3 inflammasome axis in the hippocampus of rats subjected to sleep deprivation[J]. Iran J Basic Med Sci, 2024, 27(8):1050-1058. doi:10.22038/IJBMS.2024.75068.16277.
[8]	张璐, 李莹, 吕晓丹, 等. 大鼠子宫宫腔粘连模型的建立及评价指标[J]. 动物学杂志, 2017, 52(1):115-121.
	ZHANG L, LI Y, LYU X D, et al. Establishment and evaluation of rat model with intrauterine adhesions[J]. J Zool, 2017, 52(1):115-121. doi:10.13859/j.cjz.201701013.
[9]	HATEM S, EL-KAYAL M. Novel anti-psoriatic nanostructured lipid carriers for the cutaneous delivery of luteolin:A comprehensive in-vitro and in-vivo evaluation[J]. Eur J Pharm Sci, 2023, 191:106612. doi:10.1016/j.ejps.2023.106612.
[10]	王伟, 何平, 江小明. 木犀草素及其黄酮苷的抗炎、抗氧化作用[J]. 食品科学, 2020, 41(17):208-215.
	WANG W, HE P, JIANG X M. Anti-inflammatory and antioxidant effects of luteolin and its flavone glycosides[J]. J Food Sci, 2020, 41(17):208-215. doi:10.7506/spkx1002-6630-20190908-103.
[11]	ABDELHAKIM B, DOUAE T, TAOUFIQ B, et al. Natural sources,biological effects,and pharmacological properties of cynaroside[J]. Biomed Pharmacother, 2023, 161:114337. doi:10.1016/j.biopha.2023.114337.
[12]	MENG Q L, LUO L, LEI M H, et al. Inhibition of FGFR2 signaling by cynaroside attenuates liver fibrosis[J]. Pharmaceuticals (Basel), 2023, 16(4):548. doi:10.3390/ph16040548.
[13]	XIU M X, ZHAO Y M, WANG X H, et al. Regulation of SIRT1-TLR2/TLR4 pathway in cell communication from macrophages to hepatic stellate cells contribute to alleviates hepatic fibrosis by Luteoloside[J]. Acta Histochemica, 2023, 125(1):151989. doi:10.1016/j.acthis.2022.151989.
[14]	QUAN J Y, XIE D, LI Z H, et al. Luteolin alleviates airway remodeling in asthma by inhibiting the epithelial-mesenchymal transition via β-catenin regulation[J]. Phytomedicine, 2024, 135:156090. doi:10.1016/j.phymed.2024.156090.
[15]	LIU L F, YANG H, GUO Y L, et al. The impact of chronic endometritis on endometrial fibrosis and reproductive prognosis in patients with moderate and severe intrauterine adhesions:a prospective cohort study[J]. Fertil Steril, 2019, 111(5):1002-1010.e2. doi:10.1016/j.fertnstert.2019.01.006.
[16]	ZHAO F F, WEI W, HUANG D P, et al. Knockdown of miR-27a reduces TGFβ-induced EMT and HO-induced oxidative stress through regulating mitochondrial autophagy[J]. Am J Transl Res, 2023, 15(10):6071-6082.
[17]	LIN Y F, LI Y S, CHEN P P, et al. Exosome-based regimen rescues endometrial fibrosis in intrauterine adhesions via targeting clinical fibrosis biomarkers[J]. Stem Cells Transl Med, 2023, 12(3):154-168. doi:10.1093/stcltm/szad007.
[18]	LI C Q, WANG W Y, SUN S Y, et al. Expression and potential role of MMP-9 in intrauterine adhesion[J]. Mediators Inflamm, 2021, 2021:6676510. doi:10.1155/2021/6676510.
[19]	ZHANG Z, HU J. DKK1 loss promotes endometrial fibrosis via autophagy and exosome-mediated macrophage-to-myofibroblast transition[J]. J Transl Med, 2024, 22(1):617. doi:10.1186/s12967-024-05402-5.
[20]	CUI X, XIAO H C, PAN W. The predictive value of serum IL-17A and IL-6 expression in postoperative recurrence in patients with intrauterine adhesion[J]. Am J Reprod Immunol, 2024, 91(1):e13808. doi:10.1111/aji.13808.
[21]	WU L M, ZHAN Y, WANG Y Q. Semaglutide may ameliorate fibrosis and inhibit epithelial-mesenchymal transition in intrauterine adhesion models[J]. Int J Mol Sci, 2024, 25(11):6196. doi:10.3390/ijms25116196.
[22]	WANG Y F, WANG Y X, WU Y, et al. Dulaglutide ameliorates intrauterine adhesion by suppressing inflammation and epithelial-mesenchymal transition via inhibiting the TGF-β/Smad2 signaling pathway[J]. Pharmaceuticals (Basel), 2023, 16(7):964. doi:10.3390/ph16070964.
[23]	HUANG B X, HU D, DONG A, et al. Highly antibacterial and adhesive hyaluronic acid hydrogel for wound repair[J]. Biomacromolecules, 2022, 23(11):4766-4777. doi:10.1021/acs.biomac.2c00950.
[24]	CHAE J J, JUNG J H, ZHU W, et al. Drug-free,nonsurgical reduction of intraocular pressure for four months after suprachoroidal injection of hyaluronic acid hydrogel[J]. Adv Sci (Weinh), 2020, 8(2):2001908. doi:10.1002/advs.202001908.
[25]	王沁洁, 汤彪, 谭松, 等. 交联透明质酸钠凝胶的使用时间对预防术后宫腔再粘连的疗效分析[J]. 四川医学, 2023, 44(4):380-383.
	WANG Q J, TANG B, TAN S, et al. Analysis of the efficacy of different usage time of cross-linked sodium hyaluronate for preventing postoperative intrauterine re-adhesion[J]. Sichuan Med J, 2023, 44(4):380-383.

组别		TGF-β1		MMP-9
A组		0.25±0.03		0.95±0.02
B组		0.62±0.02^a		0.26±0.04^a
C组		0.44±0.03^ab		0.36±0.04^ab
D组		0.45±0.03^ab		0.36±0.03^ab
E组		0.33±0.02^abcd		0.57±0.05^abcd
F		82.273^＊		165.654^＊
组别	Vimentin		COL5A2		COL1A1
A组	0.15±0.01		0.16±0.03		0.31±0.05
B组	0.46±0.04^a		0.69±0.07^a		0.79±0.07^a
C组	0.35±0.03^ab		0.48±0.03^ab		0.61±0.05^ab
D组	0.36±0.03^ab		0.46±0.03^ab		0.58±0.01^ab
E组	0.26±0.04^abcd		0.33±0.06^abcd		0.44±0.02^abcd
F	44.015^＊		53.979^＊		47.931^＊

组别		TGF-β1		MMP-9
A组		0.25±0.03		0.95±0.02
B组		0.62±0.02^a		0.26±0.04^a
C组		0.44±0.03^ab		0.36±0.04^ab
D组		0.45±0.03^ab		0.36±0.03^ab
E组		0.33±0.02^abcd		0.57±0.05^abcd
F		82.273^＊		165.654^＊
组别	Vimentin		COL5A2		COL1A1
A组	0.15±0.01		0.16±0.03		0.31±0.05
B组	0.46±0.04^a		0.69±0.07^a		0.79±0.07^a
C组	0.35±0.03^ab		0.48±0.03^ab		0.61±0.05^ab
D组	0.36±0.03^ab		0.46±0.03^ab		0.58±0.01^ab
E组	0.26±0.04^abcd		0.33±0.06^abcd		0.44±0.02^abcd
F	44.015^＊		53.979^＊		47.931^＊

组别	IL-6	IL-8
A组	31.08±2.59	94.10±5.99
B组	62.36±3.79^a	149.10±12.08^a
C组	52.15±1.27^ab	129.40±7.70^ab
D组	50.98±3.60^ab	131.85±4.91^ab
E组	38.02±2.04^abcd	111.09±8.05^abcd
F	96.064^＊	33.642^＊

组别	IL-6	IL-8
A组	31.08±2.59	94.10±5.99
B组	62.36±3.79^a	149.10±12.08^a
C组	52.15±1.27^ab	129.40±7.70^ab
D组	50.98±3.60^ab	131.85±4.91^ab
E组	38.02±2.04^abcd	111.09±8.05^abcd
F	96.064^＊	33.642^＊