回阳生肌膏对糖尿病大鼠阴证疮面氧化应激的影响
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摘要
目的探讨回阳生肌膏治疗糖尿病阴证疮面的作用机制。方法 108只雄性Wistar大鼠随机分为空白组18只,糖尿病组90只。将糖尿病组大鼠按照链脲佐菌素-激素干预-皮肤缺损-塑料环埋置方法制备糖尿病阴证疮面动物模型。造模成功的72只大鼠分为模型组、中药组、中药+抑制剂组、模型+抑制剂组各18只;空白组和模型组予以凡士林纱条,中药组予以回阳生肌纱条,中药+抑制剂组予以回阳生肌纱条并腹腔注射LY294002抑制剂,模型+抑制剂组予以凡士林纱条并腹腔注射LY294002抑制剂。在给药第3、7、10天检测疮面肉芽组织中磷脂酰肌醇激酶(PI3K)、蛋白激酶B (AKT)、内皮型一氧化氮合酶(eNOS)mRNA含量及超氧化物歧化酶(SOD)、丙二醛(MAD)、谷胱甘肽过氧化物酶(GSH-Px)蛋白含量。结果各组第3、7、10天大鼠疮面肉芽组织中SOD、MDA、GSH-Px含量比较,差异均有统计学意义(P <0. 001)。与空白组同时间比较,模型组第3、7、10天大鼠疮面肉芽组织中MDA、SOD和GSH-PX含量明显增高(P <0. 05)。与模型组同时间比较,中药组可显著提高大鼠疮面肉芽组织中SOD和GSH-PX含量,降低MDA含量(P <0. 05);与中药组同时间比较,中药+抑制剂组各时间SOD和GSH-PX含量均下降,MDA含量均升高(P <0. 05)。与空白组同时间相比,模型组第3、7、10天PI3K、AKT、eNOS mRNA水平均升高(P <0. 05);与模型组同时间比较,中药组第3、7、10天PI3K、AKT、eNOS mRNA水平显著提高(P <0. 05);与中药组同时间比较,中药+抑制剂组PI3K、AKT、eNOS mRNA水平下降(P <0. 05)。结论回阳生肌膏可能通过PI3K/AKT通路抑制糖尿病阴证疮面氧化应激。
Objective To investigate the effect of Huiyang Shengji Ointment(回阳生肌膏) in treating skin ulcers of diabetic rats with Yin syndrome. Methods A total of 108 male Wistar rats were randomly divided into blank group(n = 18) and diabetic group(n = 90). The diabetic rats were treated according to the method of embedding the streptozotocin hormone into the skin defect plastic ring to prepare the animal model of diabetic skin ulcer negative syndrome. The 72 successfully modeling rats were divided into a model group(n = 18),a Chinese medicine group(n =18),a Chinese medicine + inhibitor group(n = 18) and a model + inhibitor group(n = 18). The blank group and the model group were given Vaseline gauze. The Chinese medicine group was given Huiyang Shengji Ointment. The Chinese medicine + inhibitor group was given the Huiyang Shengji Ointment and intraperitoneal injection of LY294002 inhibitor. The model + inhibitor group was given Vaseline gauze and intraperitoneal injection of LY294002 inhibitor. After administration for 3 d,7 d,10 d,the phosphoinositide 3-Kinases(PI3 K),protein kinase B(AKT),endothelial nitric oxide synthase(e NOS) mRNA levels,and superoxide dismutase(SOD),malondialdehyde(MAD),glutathione peroxidase(GSH-Px) protein contents in sore granulation tissue were detected. Results The contents of SOD,MDA and GSH-Px in the sore granulation tissue on the 3 rd,7 th and 10 th day of each group were statistically significant(P < 0. 001). Compared with the blank group,the contents of MDA,SOD and GSH-Px in the sore granulation tissue of the model group were significantly increased on the 3 rd,7 th and 10 th day(P <0. 05). Compared with the model group,the Chinese medicine group can significantly increase the content of SOD and GSH-Px in the granulation tissue of the rat sore surface and reduce the MDA content(P < 0. 05). Compared with the Chinese medicine group,the SOD and GSH-Px of of the Chinese medicine + inhibitor group at each time decreased and the content of MDA increased(P < 0. 05). Compared with the blank group,the levels of PI3 K,AKT and eNOS mRNA increased on the 3 rd,7 th and 10 th day of the model group(P < 0. 05). Compared with the model group,the levels of PI3 K,AKT and eNOS mRNA the on the 3 rd,7 th and 10 th day of the Chinese medicine group.were significantly increased(P < 0. 05). Compared with the Chinese medicine group,the levels of PI3 K,AKT and eNOS mRNA in the Chinese medicine + inhibitor group decreased(P < 0. 05). Conclusion Huiyang Shengji Ointment can inhibit oxidative stress of skin ulcers of diabetic negative syndrome through the PI3 K/AKT pathway.
Objective To investigate the effect of Huiyang Shengji Ointment(回阳生肌膏) in treating skin ulcers of diabetic rats with Yin syndrome. Methods A total of 108 male Wistar rats were randomly divided into blank group(n = 18) and diabetic group(n = 90). The diabetic rats were treated according to the method of embedding the streptozotocin hormone into the skin defect plastic ring to prepare the animal model of diabetic skin ulcer negative syndrome. The 72 successfully modeling rats were divided into a model group(n = 18),a Chinese medicine group(n =18),a Chinese medicine + inhibitor group(n = 18) and a model + inhibitor group(n = 18). The blank group and the model group were given Vaseline gauze. The Chinese medicine group was given Huiyang Shengji Ointment. The Chinese medicine + inhibitor group was given the Huiyang Shengji Ointment and intraperitoneal injection of LY294002 inhibitor. The model + inhibitor group was given Vaseline gauze and intraperitoneal injection of LY294002 inhibitor. After administration for 3 d,7 d,10 d,the phosphoinositide 3-Kinases(PI3 K),protein kinase B(AKT),endothelial nitric oxide synthase(e NOS) mRNA levels,and superoxide dismutase(SOD),malondialdehyde(MAD),glutathione peroxidase(GSH-Px) protein contents in sore granulation tissue were detected. Results The contents of SOD,MDA and GSH-Px in the sore granulation tissue on the 3 rd,7 th and 10 th day of each group were statistically significant(P < 0. 001). Compared with the blank group,the contents of MDA,SOD and GSH-Px in the sore granulation tissue of the model group were significantly increased on the 3 rd,7 th and 10 th day(P <0. 05). Compared with the model group,the Chinese medicine group can significantly increase the content of SOD and GSH-Px in the granulation tissue of the rat sore surface and reduce the MDA content(P < 0. 05). Compared with the Chinese medicine group,the SOD and GSH-Px of of the Chinese medicine + inhibitor group at each time decreased and the content of MDA increased(P < 0. 05). Compared with the blank group,the levels of PI3 K,AKT and eNOS mRNA increased on the 3 rd,7 th and 10 th day of the model group(P < 0. 05). Compared with the model group,the levels of PI3 K,AKT and eNOS mRNA the on the 3 rd,7 th and 10 th day of the Chinese medicine group.were significantly increased(P < 0. 05). Compared with the Chinese medicine group,the levels of PI3 K,AKT and eNOS mRNA in the Chinese medicine + inhibitor group decreased(P < 0. 05). Conclusion Huiyang Shengji Ointment can inhibit oxidative stress of skin ulcers of diabetic negative syndrome through the PI3 K/AKT pathway.
引文
[1]李媛.糖尿病慢性皮肤溃疡阴证模型建立及回阳生肌膏对糖尿病大鼠阴证溃疡创面不同时相的影响[D].北京:北京中医药大学,2014:29-35.
[2]吴海霞.氧化应激反应在糖尿病慢性皮肤溃疡中的作用[J].实用临床医药杂志,2016,20(5):185-187.
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[5]王广宇,董建勋,徐旭英,等.回阳生肌散对慢性难愈性皮肤溃疡伴铜绿假单胞菌感染31例[J].辽宁中医杂志,2013,40(10):2045-2048.
[6]HUANG X,LIU G,GUO J,et al.The PI3K/AKT pathway in obesity and type 2 diabetes[J].Int J Biol Sci,2018,14(11):1483-1496.
[7]付小兵.糖尿病慢性难愈合创面大鼠模型的制备[J].上海实验动物科学,1997,17(4):217-219.
[8]MIRANDA-DAZ AG,PAZARN-VILLASEOR L,YANOWSKY-ESCATELL FG,et al.Oxidative stress in diabetic nephropathy with early chronic kidney disease[J].J DIABETES RES,2016:7047238.doi:10.1155/2016/7047238.
[9]LI W,KANDHARE AD,MUKHERJEE AA,et al.Hesperidin,a plant flavonoid accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats:role of TGF-β/Smads and Ang-1/Tie-2 signaling pathways[J].EXCLI J,2018,17:399-419.doi:10.17179/excli2018-1036.
[10]刘继松,方勇.氧化应激和糖尿病创面的延迟愈合[J].蚌埠医学院学报,2010,35(7):756-757.
[11]赵光明,董建勋,李媛,等.回阳生肌散对糖尿病皮肤溃疡大鼠创面血管生成相关因子的影响[J].中医杂志,2014,55(6):504-507.
[12]李媛,赵光明,董建勋,等.回阳生肌膏及其拆方对糖尿病慢性皮肤溃疡大鼠血清白介素-1α和血栓素B2含量的影响[J].中国中医药信息杂志,2014,21(7):43-46.
[13]MA L,LI P,SHI Z,et al.A prospective,randomized,controlled study of hyperbaric oxygen therpy:effects on healing and oxidative stress of ulcer tissue in patients with a diabetic foot ulcer[J].Ostomy Wound Manage,2013,59(3):18-24.
[14]ASHAFAQ M,VARSHNEY L,KHAN MH,et al.Neuromodulatory effects of hesperidin in mitigating oxidative stress in streptozotocin induced diabetes[J].Biomed Res Int,2014:249031.doi:10.1155/2014/249031.
[15]AN ZM,DONG XG,GUO Y,et al.Effects and clinical significance of pentoxifylline on the oxidative stress of rats with diabetic nephropathy[J].J Huazhong Univ Sci Technolog Med Sci,2015,35(3):356-361.
[16]VAIRAMON SJ,BABU M,VISWANATHAN V.Oxidative stress markers regulating the healing of foot ulcers in patients with type 2 diabetes[J].Wounds,2009,21(10):273-279.
[2]吴海霞.氧化应激反应在糖尿病慢性皮肤溃疡中的作用[J].实用临床医药杂志,2016,20(5):185-187.
[3]贾连城,盛巡,孙明杰,等.回阳生肌膏治疗慢性难愈性皮肤溃疡疗效观察及作用机制探讨[J].中国中医药信息杂志,2009,16(10):10-12.
[4]谢志钩.回阳生肌膏治疗慢性皮肤溃疡临床研究[J].亚太传统医药,2015,11(17):123-124.
[5]王广宇,董建勋,徐旭英,等.回阳生肌散对慢性难愈性皮肤溃疡伴铜绿假单胞菌感染31例[J].辽宁中医杂志,2013,40(10):2045-2048.
[6]HUANG X,LIU G,GUO J,et al.The PI3K/AKT pathway in obesity and type 2 diabetes[J].Int J Biol Sci,2018,14(11):1483-1496.
[7]付小兵.糖尿病慢性难愈合创面大鼠模型的制备[J].上海实验动物科学,1997,17(4):217-219.
[8]MIRANDA-DAZ AG,PAZARN-VILLASEOR L,YANOWSKY-ESCATELL FG,et al.Oxidative stress in diabetic nephropathy with early chronic kidney disease[J].J DIABETES RES,2016:7047238.doi:10.1155/2016/7047238.
[9]LI W,KANDHARE AD,MUKHERJEE AA,et al.Hesperidin,a plant flavonoid accelerated the cutaneous wound healing in streptozotocin-induced diabetic rats:role of TGF-β/Smads and Ang-1/Tie-2 signaling pathways[J].EXCLI J,2018,17:399-419.doi:10.17179/excli2018-1036.
[10]刘继松,方勇.氧化应激和糖尿病创面的延迟愈合[J].蚌埠医学院学报,2010,35(7):756-757.
[11]赵光明,董建勋,李媛,等.回阳生肌散对糖尿病皮肤溃疡大鼠创面血管生成相关因子的影响[J].中医杂志,2014,55(6):504-507.
[12]李媛,赵光明,董建勋,等.回阳生肌膏及其拆方对糖尿病慢性皮肤溃疡大鼠血清白介素-1α和血栓素B2含量的影响[J].中国中医药信息杂志,2014,21(7):43-46.
[13]MA L,LI P,SHI Z,et al.A prospective,randomized,controlled study of hyperbaric oxygen therpy:effects on healing and oxidative stress of ulcer tissue in patients with a diabetic foot ulcer[J].Ostomy Wound Manage,2013,59(3):18-24.
[14]ASHAFAQ M,VARSHNEY L,KHAN MH,et al.Neuromodulatory effects of hesperidin in mitigating oxidative stress in streptozotocin induced diabetes[J].Biomed Res Int,2014:249031.doi:10.1155/2014/249031.
[15]AN ZM,DONG XG,GUO Y,et al.Effects and clinical significance of pentoxifylline on the oxidative stress of rats with diabetic nephropathy[J].J Huazhong Univ Sci Technolog Med Sci,2015,35(3):356-361.
[16]VAIRAMON SJ,BABU M,VISWANATHAN V.Oxidative stress markers regulating the healing of foot ulcers in patients with type 2 diabetes[J].Wounds,2009,21(10):273-279.