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HucMSC-Ex Mediates Ca2+ Influx and Promotes Diabetic Wound Healing through Regulating TRPC6


WANG Min1,2, WU Peipei3,4, SHI Hui1 *

(1 Jiangsu Key Laboratory of Medical Science and Laboratory Medicine, Department of Clinical Laboratory, School of Medicine, Jiangsu University, Zhenjiang 212000, China; 2 School of Biomedical Engineering, Anhui Medical University, Hefei 230032, China; 3 Department of Clinical Laboratory, the First Affiliated Hospital of USTC, Hefei 230000, China; 4 Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230000, China)
Abstract:

t This study aimed to investigate the changes and functions of TRPC6 (transient potential canonical 6) in diabetic wounds and the regulatory role of hucMSC-Ex (human umbilical cord mesenchymal stem cell derived exosome) on TRPC6. Tissue immunofluorescence was used to detect the expression of TRPC6 in clinical patients with DFU (diabetic foot ulcer). qRT-PCR and Western blot were used to detect the changes of TRPC6 mRNA and protein expression in rat DFs (dermal fibroblasts) after stimulation with AGE-BSA (bovine serum albumin modified with advanced glycosylation end products), respectively. Moreover, DFs were transfected with siRNA (small interfering RNA) and overexpressed plasmid to investigate the effects of TRPC6 on the biological functions of DFs, such as Ca2+ influx, proliferation, and collagen secretion. Cellular immunofluorescence was used to detect the effect of hucMSC-Ex on the ability of DFs to secrete collagen. A DFU model was constructed in SD rats, and hucMSC-Ex was added to intervene, and the skin structure was observed by HE staining, to evaluate the therapeutic effect of hucMSC-Ex on DFU. Furthermore, qRT-PCR, Western blot, cellular immunofluorescence, and immunohistochemical staining were used to detect the effect of hucMSC-Ex on TRPC6. The results showed that TRPC6 channel protein was significantly decreased in diabetic wounds. Knock-down of TRPC6 decreased Ca2+ influx and impaired the biological function of DFs. On the contrary, overexpression of TRPC6 enhanced Ca2+ influx and biological function of DFs. In conclusion, hucMSC-Ex could regulate TRPC6-mediated Ca2+ influx promote diabetic wound healing.


CSTR: 32200.14.cjcb.2024.02.0004