基于PINK1/Parkin通路介导的线粒体自噬 探讨中药丹黄散对糖尿病溃疡的促愈机制.

王庚新; 张春玲; 赵伟; 陈露; 邸铁涛; 张云; 王艳辉; 刘海艳

To explore the promotion mechanism of Danhuang Powder on the wound of diabetic ulcers by taking the mitophagy through the PINK1/Parkin pathway as a target. Method:18 specific pathogen free（SPF） grade male db/db mice and 6 SPF-grade male db/m mice with the same genetic background were selected. The 18 db/db mice were divided into the model group, growth factor group, and Danhuang Powder group, 6 in each, according to the random digital tables method;6 db/m mice as a blank group. Use 4% chloral hydrate for anesthesia, and establish a skin defect with a diameter of 0. 8 cm on the back of mice with a sterile pitch to prepare the diabetic ulcer model. The blank group and the model group received non - drug intervention. The control group and the Danhuang Powder group received the restructured human epidermal growth factor gel and Danhuang Powder for external application, respectively. The medicine was changed once a day for 21 d. On the 7th and 14th days after the model creation, the facade organization analysis was taken to analyze. The pathological situation of wounds was oberved through HE and Masson staining. The condition of wound tissue mitochondria and spontaneous bodies through transmitted electron microscope;detect wound tissue LC3-Ⅰ and LC3-Ⅱ, PINK1, and Parkin protein expression were observed by Western-Blot. Results:The healing rate of the traumatic surface of the blank group, growth factor group, and Danhuang San group was higher than that of the model group on the 7th, 14th, and 21st days after modeling（P<0. 05）. HE and Masson staining of the traumatic surface of the mice on the 7th and 14th days showed that compared with the model group, the inflammatory reaction of the blank group, growth factor group, and Danhuang Powder group was lighter; the number of neovascularization increased significantly;the cells in the traumatic surface were closely arranged;the traumatic stroma formed abundant collagen fibers, and the traumatic epidermal tissue recovered well. Transmission electron microscopic observation of the traumatic tissue on day 7 revealed that the mitochondrial swelling condition in the blank group, the growth factor group, and the Danhuang Powder group were lighter than the model group, which showed that the quantity of autophagosome was higher than the model group;the Western-Blot test showed that LC3-Ⅱ/LC3-Ⅰ, PINK1, and Parkin protein expression of the blank group, growth factor group, and Danhuang Powder group were higher than the model group（P<0. 05）. Conclusions:Danhuang Powder could raise the PINK1/Parkin pathway and active mitophagy to protect the normal structure and functions of mitochondria in wound cells, promote the growth of vascular and granulite tissue, and effectively promote ulcer wound healing in diabetic mice.

PROTEIN kinases; WOUND healing; BIOLOGICAL models; COLLAGEN; HERBAL medicine; STAINS & staining (Microscopy); DIABETIC foot; AUTOPHAGY; ANIMAL experimentation; EPIDERMAL growth factor; WESTERN immunoblotting; NEOVASCULARIZATION; ORGANELLES; CYTOSKELETAL proteins; MITOCHONDRIA; CELLULAR signal transduction; ELECTRON microscopy; GRANULATION tissue; EPIDERMIS; CHINESE medicine; MICE; RECOMBINANT proteins

Chinese Nursing Research, 2022, Vol 36, Issue 24, p4370

1009-6493

Academic Journal

10.12102/j.issn.1009-6493.2022.24.008