回阳生肌膏对糖尿病大鼠阴证疮面氧化应激的影响

Effects of Huiyang Shengji Ointment( 回阳生肌膏) on the Oxidative Stress of the Ulcer Surface with Yin Syndrome of Diabetes Mellitus Rats

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 ( PI3K),protein kinase B ( AKT),endothelial nitric oxide synthase ( eNOS) 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 3rd,7th and 10th 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 3rd,7th and 10th 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 PI3K,AKT and eNOS mRNA increased on the 3rd,7th and 10th day of the model group ( P < 0. 05). Compared with the model group,the levels of PI3K,AKT and eNOS mRNA the on the 3rd,7th and 10th day of the Chinese medicine group. were significantly increased ( P < 0. 05). Compared with the Chinese medicine group,the levels of PI3K,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 PI3K /AKT pathway.