人参皂苷Rg3对糖尿病大鼠难愈创面表皮细胞及血管新生的影响
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摘要
目的从改善皮肤易损状态方面,探讨人参皂苷Rg3(ginsenoside Rg3, Rg3)修复糖尿病难愈创面的作用机制。方法♂SD大鼠制备糖尿病模型,随机分为模型组、氨基胍(aminoguanidirig, AG)组、Rg3高、低剂量组,另设正常组。灌胃4周后,背部皮肤等面积全层切除,伤后d 21创面皮肤取材,观测病理形态及表皮、真皮层厚度;分析表皮细胞增殖周期;检测CD31表达。结果 Rg3给药组成纤维细胞较多,坏死组织脱落及上皮匍行提前,表皮及真皮层增厚(P<0.01);表皮细胞增殖周期,模型组G_0/G_1期比例升高(P<0.01),G_2/M、S期比例降低(P<0.01),AG、Rg3组G_0/G_1期比例下降,且高剂量下降明显(P<0.05),G_2/M、S期比例升高(P<0.05或P<0.01);模型组CD31蛋白表达明显降低(P<0.01),AG、Rg3高剂组表达升高(P<0.05)。结论 Rg3通过改善皮肤病理状态,调节表皮细胞增殖周期和促进血管生成,有效修复糖尿病性皮肤损伤。
Aim To explore the repair mechanism of ginsenoside Rg3 in treating the impaired diabetic wound from improving the vulnerability of diabetic skin of rats. Methods Male SD rats(n=32) were injected intraperitoneally streptozotocin to induce hyperglycemia, except for the rats of control group(n=8). Diabetic rats were randomly divided into model group(equal volume of saline), aminoguanidine group(10 mg·kg~(-1)), ginsenoside Rg3 high dose(15 mg·kg~(-1)) and low group(5 mg·kg~(-1)). After four weeks of oral gavage treatment, full-thickness wound was established. On 21 st day after injury, wound samples were collected to observe the pathological changes in wound; the thickness of epidermis and dermis was measured by HE staining; and the epidermal cell proliferation cycle was analysed by flow cytometry; the expression of CD31 in wound tissues was detected by immunohistochemical and image analytical methods. Results Ginsenoside Rg3 groups showed significantly more fibroblasts, necrotic tissue drops and advanced epithelial, and thickening of epidermis and dermis(P<0.01). Model group showed significant increase in G_0/G_1 phase ratio of epidermal cell cycle( P < 0. 01),while reduction in G_2/M and S period ratios( P < 0. 01).However,G_0/G_1 period ratio decreased,while G_2/M and S period ratios rose in aminoguanidine group,ginsenoside Rg3 high dose and low dose groups. The decrease of G_0/G_1 period ratio( P < 0. 05) and increase in G2/M( P < 0. 05) and S period ratios( P < 0. 01)was found to be significant. The expressions of CD31 in model group was lower than those in control group( P < 0. 01). Whereas,it was higher in ginsenoside Rg3 high dose group and aminoguanidine group( P <0. 05). Conclusions Ginsenoside Rg3 can effectively promote the repair and healing of impaired diabetic wound. The various mechanisms of repair might be through improving skin pathology,regulating epidermal cell proliferation cycle and promoting angiogenesis.
Aim To explore the repair mechanism of ginsenoside Rg3 in treating the impaired diabetic wound from improving the vulnerability of diabetic skin of rats. Methods Male SD rats(n=32) were injected intraperitoneally streptozotocin to induce hyperglycemia, except for the rats of control group(n=8). Diabetic rats were randomly divided into model group(equal volume of saline), aminoguanidine group(10 mg·kg~(-1)), ginsenoside Rg3 high dose(15 mg·kg~(-1)) and low group(5 mg·kg~(-1)). After four weeks of oral gavage treatment, full-thickness wound was established. On 21 st day after injury, wound samples were collected to observe the pathological changes in wound; the thickness of epidermis and dermis was measured by HE staining; and the epidermal cell proliferation cycle was analysed by flow cytometry; the expression of CD31 in wound tissues was detected by immunohistochemical and image analytical methods. Results Ginsenoside Rg3 groups showed significantly more fibroblasts, necrotic tissue drops and advanced epithelial, and thickening of epidermis and dermis(P<0.01). Model group showed significant increase in G_0/G_1 phase ratio of epidermal cell cycle( P < 0. 01),while reduction in G_2/M and S period ratios( P < 0. 01).However,G_0/G_1 period ratio decreased,while G_2/M and S period ratios rose in aminoguanidine group,ginsenoside Rg3 high dose and low dose groups. The decrease of G_0/G_1 period ratio( P < 0. 05) and increase in G2/M( P < 0. 05) and S period ratios( P < 0. 01)was found to be significant. The expressions of CD31 in model group was lower than those in control group( P < 0. 01). Whereas,it was higher in ginsenoside Rg3 high dose group and aminoguanidine group( P <0. 05). Conclusions Ginsenoside Rg3 can effectively promote the repair and healing of impaired diabetic wound. The various mechanisms of repair might be through improving skin pathology,regulating epidermal cell proliferation cycle and promoting angiogenesis.
引文
[1] Long M, Rojo de la Vega M, Wen Q, et al. An essential role of NRF2 in diabetic wound healing[J]. Diabetes, 2016,65(3):780-93.
[2] 李静平, 郑永仁, 张燕.人参皂苷Rg3对糖尿病大鼠创面修复作用的实验研究[J].云南中医中药杂志, 2016,37(1):25-8.[2] Li J P, Zheng Y R, Zhang Y. Experimental study of ginsenoside Rg3 on wound repair in diabetic rats[J]. Yunnan J Tradit Chin Med, 2016,37(1):25-8.
[3] 张颖, 邢伟, 黄宏,等.2型糖尿病非收缩性难愈伤口模型的建立[J].中国组织工程研究, 2012,16(24):4432-6.[3] Zhang Y, Xing W, Huang H,et al. Establishment of a non-contractile refractory wound model in type 2 diabetic rats[J]. Chin J Tissue Eng Res, 2012,16(24):4432-6.
[4] 杨选鑫, 惠琦, 曹高忠,等. rh-aFGF卡波姆940凝胶对1型糖尿病大鼠皮肤创伤的修复作用[J].中国药理学通报, 2018,34(4):557-62.[4] Yang X X, Hui Q, Cao G Z, et al. The therapeutic effect of rh-aFGF carbomer 940 gel on wound healing in type I diabetic SD rat models[J]. Chin Pharmacol Bull, 2018,34(4):557-62.
[5] 杨秀颖, 杜冠华.糖基化终末产物及相关药物研究进展[J].中国药理学通报, 2011,27(9):1185-8.[5] Yang X Y,Du G H. Advanced glycosylation end products and related drug development[J]. Chin Pharmacol Bull, 2011,27(9):1185-8.
[6] Takahashi P Y, Kiemele L J, Chandra A, et al. A retrospective cohort study of factors that affect healing in long-term care residents with chronic wounds[J]. Ostomy Wound Manage, 2009,55(1):32-7.
[7] 赵文杰, 陈迪, 倪劲松,等. 20(S)-人参皂苷Rg3对前列腺癌 PC-3M细胞的诱导凋亡作用[J].中国药理学通报, 2009,25(2):235-8.[7] Zhao W J, Chen D, Ni J S, et al. Study on PC-3M cell apoptosis induced by 20(s)-ginsendoside Rg3 in prostate carcinoma[J]. Chin Pharmacol Bull, 2009,25(2):235-8.
[8] 陆树良, 谢挺, 牛轶雯,等.糖尿病合并创面难愈的机制研究[J].药品评价, 2011,8(7):17-21.[8] Lu S L, Xie T, Niu Y W, et al. Mechanism research of diabetic wound hard to be cured[J]. Drug Evaluation, 2011,8(7):17-21.
[9] Xu J, Carlos Zgheib C, Hodges M M, et al. Mesenchymal stem cells correct impaired diabetic wound healing by decreasing ECM proteolysis[J]. Physiol Genomics, 2017,49(10):541-8.
[10] 刘克辉,许力军,王玲,等.金因肽局部治疗糖尿病皮肤感染的临床观察[J].中国药理学通报, 2005,21(12):1535-6.[10] Liu K H, Xu L J, Wang L, et al. Observing the clinical effect of rhEGF for treating diabetes patients skin infection[J]. Chin Pharmacol Bull,2005,21(12):1535-6.
[11] Dulmovits B M, Hermam I M. Microvascular remodeling and wound healing:a role for pericytes[J]. Int J Biochem Cell Biol, 2012,44(11):1800-12.
[12] Reid B, Zhao M. The electrical response to injury:molecular mechanisms and wound healing[J]. Adv Wound Care, 2014,3(2):184-201.
[13] Patrick Y K, Daisy Y L, Wu P K, et al. The angiosupp-ressive effects of 20(R)-ginsenoside Rg3[J]. Biochem Pharmacol,2006,72(4):437-45.
[2] 李静平, 郑永仁, 张燕.人参皂苷Rg3对糖尿病大鼠创面修复作用的实验研究[J].云南中医中药杂志, 2016,37(1):25-8.[2] Li J P, Zheng Y R, Zhang Y. Experimental study of ginsenoside Rg3 on wound repair in diabetic rats[J]. Yunnan J Tradit Chin Med, 2016,37(1):25-8.
[3] 张颖, 邢伟, 黄宏,等.2型糖尿病非收缩性难愈伤口模型的建立[J].中国组织工程研究, 2012,16(24):4432-6.[3] Zhang Y, Xing W, Huang H,et al. Establishment of a non-contractile refractory wound model in type 2 diabetic rats[J]. Chin J Tissue Eng Res, 2012,16(24):4432-6.
[4] 杨选鑫, 惠琦, 曹高忠,等. rh-aFGF卡波姆940凝胶对1型糖尿病大鼠皮肤创伤的修复作用[J].中国药理学通报, 2018,34(4):557-62.[4] Yang X X, Hui Q, Cao G Z, et al. The therapeutic effect of rh-aFGF carbomer 940 gel on wound healing in type I diabetic SD rat models[J]. Chin Pharmacol Bull, 2018,34(4):557-62.
[5] 杨秀颖, 杜冠华.糖基化终末产物及相关药物研究进展[J].中国药理学通报, 2011,27(9):1185-8.[5] Yang X Y,Du G H. Advanced glycosylation end products and related drug development[J]. Chin Pharmacol Bull, 2011,27(9):1185-8.
[6] Takahashi P Y, Kiemele L J, Chandra A, et al. A retrospective cohort study of factors that affect healing in long-term care residents with chronic wounds[J]. Ostomy Wound Manage, 2009,55(1):32-7.
[7] 赵文杰, 陈迪, 倪劲松,等. 20(S)-人参皂苷Rg3对前列腺癌 PC-3M细胞的诱导凋亡作用[J].中国药理学通报, 2009,25(2):235-8.[7] Zhao W J, Chen D, Ni J S, et al. Study on PC-3M cell apoptosis induced by 20(s)-ginsendoside Rg3 in prostate carcinoma[J]. Chin Pharmacol Bull, 2009,25(2):235-8.
[8] 陆树良, 谢挺, 牛轶雯,等.糖尿病合并创面难愈的机制研究[J].药品评价, 2011,8(7):17-21.[8] Lu S L, Xie T, Niu Y W, et al. Mechanism research of diabetic wound hard to be cured[J]. Drug Evaluation, 2011,8(7):17-21.
[9] Xu J, Carlos Zgheib C, Hodges M M, et al. Mesenchymal stem cells correct impaired diabetic wound healing by decreasing ECM proteolysis[J]. Physiol Genomics, 2017,49(10):541-8.
[10] 刘克辉,许力军,王玲,等.金因肽局部治疗糖尿病皮肤感染的临床观察[J].中国药理学通报, 2005,21(12):1535-6.[10] Liu K H, Xu L J, Wang L, et al. Observing the clinical effect of rhEGF for treating diabetes patients skin infection[J]. Chin Pharmacol Bull,2005,21(12):1535-6.
[11] Dulmovits B M, Hermam I M. Microvascular remodeling and wound healing:a role for pericytes[J]. Int J Biochem Cell Biol, 2012,44(11):1800-12.
[12] Reid B, Zhao M. The electrical response to injury:molecular mechanisms and wound healing[J]. Adv Wound Care, 2014,3(2):184-201.
[13] Patrick Y K, Daisy Y L, Wu P K, et al. The angiosupp-ressive effects of 20(R)-ginsenoside Rg3[J]. Biochem Pharmacol,2006,72(4):437-45.