Shixiang Paste interferes with the expression of advanced glycation endproducts and its receptors and endothelial nitric oxide synthase in wound tissue of diabetic ulcer rats

doi:10.12307/2022.620

Abstract

Abstract: BACKGROUND: Diabetic ulcer, as one of the common complications of diabetes, has caused serious harm to human health due to its low healing rate and high amputation rate. Although traditional Chinese medicine has been proved to improve wound healing, the molecular mechanism of this effect has not been clarified.
OBJECTIVE: To investigate the possible mechanism of Shixiang Paste in treating diabetic ulcers.
METHODS: Forty clean male Sprague-Dawley rats were randomly divided into four groups (n=10 per group): a normal control group, a diabetic ulcer group, a cell growth factor group and a Shixiang Paste group. Except for the normal control group, a diabetic rat model was established in the other three groups. After 1 week, an ulcer wound model was established in four groups by using the full-thickness skin defect method. One week after successful modeling, rats in the Shixiang Paste and cell growth factor groups were treated with external Shixiang Paste and cell growth factor, respectively. Wounds in the diabetic ulcer and normal control groups were only covered and fixed with double sterile dry gauze. The granulation tissue samples were taken from the wound on the back of rats on the 7th and 14th days after intervention. Western blot was used to detect the protein expression of receptors of advanced glycation endproducts and endothelial nitric oxide synthase in the granulation tissue from the back wound of rats. Hematoxylin-eosin staining was used to detect pathological changes in rat back wound samples. Immunohistochemical staining was used to detect the expression of advanced glycation endproducts in the granulation tissue from the wound on the back of rats.
RESULTS AND CONCLUSION: The results of hematoxylin-eosin staining showed that large areas of unrepaired tissue were seen under the microscope in the normal control group and the diabetic ulcer group on the 7th and 14th days after intervention, while in the Shixiang Paste and cell growth factor groups, the wounds healed well on the 7th and 14th days after intervention. Western blot results showed that, compared with the normal control group, the expression of receptors of advanced glycation endproducts was decreased in the diabetic ulcer group was increased, and the expression of endothelial nitric oxide synthase (P < 0.05). In addition, the expression of receptors of advanced glycation endproducts was relatively decreased, and the expression of endothelial nitric oxide synthase was relatively increased in the Shixiang Paste group and cell growth factor group (P < 0.05). Compared with the diabetic ulcer group, the expression of receptors of advanced glycation endproducts was relatively decreased, and the expression of endothelial nitric oxide synthase was relatively increased in the Shixiang Paste group and the cell growth factor group (P < 0.05). Compared with the cell growth factor group, the expression of receptors of advanced glycation endproducts was relatively decreased, and the expression of endothelial nitric oxide synthase was relatively increased in the Shixiang Paste group (P < 0.05). The results of immunohistochemical staining showed that, compared with the normal control group, the expression of advanced glycation endproducts was increased in the diabetic ulcer group and the cell growth factor group, and the expression of advanced glycation endproducts was decreased in the Shixiang Paste group (P < 0.05). Compared with the diabetic ulcer group, the expression of advanced glycation endproducts was increased in the other groups (P < 0.05). Compared with the Shixiang Paste group, the expression level of advanced glycation endproducts was decreased in the cell growth factor group (P < 0.05). To conclude, Shixiang Paste can promote the repair of diabetic ulcer wound. Inhibiting the expression of advanced glycation endproducts and receptors and enhancing the expression of endothelial nitric oxide synthase in wound tissue may be one of the mechanisms.

Key words: Shixiang Paste, diabetic ulcer, advanced glycation endproduct, receptor of advanced glycation endproduct, endothelial nitric oxide synthase

CLC Number:

Fei Ji, Zhang Kaiwei, Zhou Yifu, Ling Yiming. Shixiang Paste interferes with the expression of advanced glycation endproducts and its receptors and endothelial nitric oxide synthase in wound tissue of diabetic ulcer rats[J]. Chinese Journal of Tissue Engineering Research, 2022, 26(20): 3196-3201.

Figures/Tables 8

References

[1] JAWORSKI M, PANCZYK M, SLIWCZYRISKI A, et al. Severe depressive episode with psychotic symptoms and type 2 diabetes: a 2010-2017 longitudinal study. Med Sci Monit. 2019;25:1760-1768.
[2] ADU MD, MALABU UH, MALAU-ADULI AEO, et al. Enablers and barriers to effective diabetes self-management: a multi-national investigation. PLoS One. 2019;14:e0217771.
[3] INFANTE-GARCIA C, GARCIA-ALLOZA M. Review of the effect of natural compounds and extracts on neurodegeration in Animal models of diabetes mellitus. Int J Mol Sci. 2019;20:E2533.
[4] YANG WY. Epidemiology and trends in diabetes in China (in Chinese). Sci Sin Vitae. 2018;48:812-819.
[5] CUNNION KM, KRISHNA NK, PALLERA HK, et al. Complement activation and STAT4 expression are associated with early inflammation in diabetic wounds. PloS One. 2017;12(1):e0170500.
[6] WOZNIAK SE, GEE LL, WACHTEL MS, et al. Adipose Tissue: The New Endocrine Organ A Review Article. Dig Dis Sci. 2009; 54(9): 1847-1856.
[7] SCHAFER M, WERNER S. Oxidative stress in normal and impaired wound repair. Pharmacol Res. 2008;58:165-167.
[8] KOENEN P,SPANHOLTZ TA,MAEGELE M, et al. Acute and chronic wound fluids inversely influence adi- pose-derived stem cell function: molecular insights into impaired wound healing. Int Wound J. 2015;12(1):10-16.
[9] BOLLA SR,MOHAMMED AL-SUBAIE A,YOUSUF AL-JINDAN R. In vitro wound healing potency of metha- nolic leaf extract of Aristolochia saccata is possibly media- ted by its stimulatory effect on collagen-1 expression. Heliyon. 2019;5(5): e01648.
[10] 凌一鸣,费冀,张开伟,等. 石香膏对糖尿病慢性难愈合创面模型大鼠创面组织血管内皮细胞黏附分子1、核因子κB p65 表达的影响[J]. 中医杂志,2020, 61(7): 619-625.
[11] WAUTIER MP, GUILLAUSSEAU PJ, WAUTIER JL. Activation of the receptor for advanced glycation end products and consequences on health. Diabetes Metab Syndr. 2017;11(4):305-309.
[12] 李文华,刘筱,周雯婷,等. 糖尿病溃疡动物模型的研究进展[J]. 湖南中医杂志,2018,34(8):246-248.
[13] 卓燊,乔雪,陈君,等. 复方愈疡散治疗糖尿病慢性难愈性创面的机制[J]. 中国实验方剂学杂志,2015,21(4):115-119.
[14] 陆少君,曾伟斌,臧林泉. 2 型糖尿病大鼠模型制备实验研究[J]. 广东药科大学学报,2017,33(5):624-628
[15] 凌一鸣,费冀,张开伟,等. 中药石香膏对糖尿病大鼠糖尿病溃疡 RAGE/NF-κBp65/eNOS mRNA表达影响的研究[J]. 中国中医基础医学杂志,2019,25(7): 913-917.
[16] 吴允波,胡铁山,程仕萍,等. 解毒祛腐煨脓生肌法治疗蛇伤后溃疡的临床研究[J]. 时珍国医国药,2018,29(6):1382-1384.
[17] 韩璐,孙甲友,周丽,等. 没药化学成分和药理作用研究进展[J]. 亚太传统医药,2015,11(3):38-42.
[18] 周灵君,张丽,丁安伟. 炉甘石敛口生肌的药效学研究[J]. 中药新药与临床药理,2013,24(4):333-337.
[19] 尚坤,李敬文,常美月,等. 冰片化学成分及药理作用研究[J]. 吉林中医药, 2018,38(1):93-95.
[20] KANG R, CHEN R, XIE M, et al. The receptor for advanced glycation end products activates the AIM2 inflammasome in acute pancreatitis. Immunol. 2016;196(10): 4331-4337.
[21] ZHANG J, YAN J. Protective effect of ginkgolic acid in attenuating LDL induced inflammation human peripheral blood mononuclear cells via altering the NF-κB signaling pathway. Front Pharmacol. 2019;8(10):1241-1252.
[22] CONG L, YANG S, ZHANG Y, et al. DFMG attenuates the activation of macrophages induced by co-culture with LPC-injured HUVE-12 cells via the TLR4/MyD88/NF-κB signaling pathway. Int J Mol Med. 2018;41(5):2619-2628.
[23] IWAKIRI Y. S-nitrosylation of proteins: a new insight into endothelial cell function regulated by eNOS-devived NO. Nitric Oxide. 2011;25(2):95-101.
[24] Chen Z, DS Oliveira S, Zimnicka AM. Reciprocal Regulation of eNOS and Caveolin-1 Functions in Endothelial Cells. Mol Biol Cell. 2018;29(10):1190-1202.
[25] Papapetropoulos A, Carcina-Cardena G, Madri JA, et al. Nitric oxide production contributes to the angiogenic properties of vascular endothelial growth factor in humanendothelial cells. Clin Invest. 1997;100:3131-3139.
[26] SCHIEKOFER S, ANDRASSY M, CHEN J, et al. Acute hyperglycemia causesintracellular formation of CML,and activation of ras,p42/p44MAPK,and nuclear factor-kB in PBMCs. Diabetes. 2003;52:621-633.
[27] 宋涛,吴树金,刘玉晖,等.Cariporide对糖基化终末产物所致血管内皮损伤的保护作用[J]中南药学,2007,5(5):429-432.
[28] BIERHAUS A, HUMPERT PM, MORCOS M, et al. Understanding RAGE,the receptor for advanced glycation end products. Mol Med. 2005;83(11):876-886.
[29] IWAKIRI Y. S-nitrosylation of proteins: a new insight into endothelial cell function regulated by eNOS-devived NO. Nitric Oxide. 2011;25(2):95-101.
[30] CHEN Z, DS OLIVEIRA S, ZIMNICKA AM. Reciprocal Regulation of eNOS and Caveolin -1 Functions in Endothelial Cells. Mol Biol Cell. 2018;29(10):1190-1202.