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| Effect of Danzhi Jiangtang Capsule on regulating β-catenin protein on high phosphorus-induced calcification of vascular endothelial cell in lower limb in diabetic mice |
| Hits 1257 Download times 1065 Received:October 10, 2022 |
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| DOI
10.11656/j.issn.1673-9043.2023.01.16 |
| Key Words
diabetes;Danzhi Jiangtang Capsule;β-catenin;calcification of endothelial cell |
| Author Name | Affiliation | | NI Yingqun | The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China | | FANG Zhaohui | The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China | | LI Juyi | The First Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230031, China | | SHI Hui | Anhui University of Traditional Chinese Medicine, Hefei 230038, China | | YU Dandan | Anhui University of Traditional Chinese Medicine, Hefei 230038, China | | LIU Guangju | Anhui University of Traditional Chinese Medicine, Hefei 230038, China | | CHEN Wenjuan | Anhui University of Traditional Chinese Medicine, Hefei 230038, China |
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| Abstract
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| [Objective] To investigate the regulating effect of Danzhi Jiangtang Capsule on high phosphorus-induced calcification of vascular endothelial cell in lower limb in diabetic mice.[Methods] Endothelial cells were isolated from femoral artery of diabetic mice and healthy mice and both cultured in vitro. They were divided into blank group,high phosphorus group,Danzhi group and LiCL group. After adjusting the stable state of EC cells,except for the blank group,the other groups were added with high phosphorus medium (containing 10 mmol/L β-glycerophosphate-50 mg/mL vitamin C and 1×10-7m insulin) to induce cell calcification. Meanwhile,Danzhi group and LiCL group were given 10% drug-containing serum (Danzhi hypoglycemic capsule) for 48 h. LiCL group was given 20 mmol/L LiCL for further intervention for 48 h.Cell apoptosis was detected by TUNEL assay and cell proliferation was detected by MTT assay.The mRNA expression levels of α-SMA,β-catenin,alkaline phosphatase (ALP),osteopontin (OPN),bone morphogenetic protein 2(BMP2) and curly homolog 1(FZD1) were detected by reverse transcription-polymerase chain reaction (RT-PCR). The localization of β-catenin protein was observed by confocal laser microscopy.[Results] Compared with blank group,apoptosis number,Ca2+ content,expression levels of α-SMA,β-catenin,ALP,OPN,BMP2 and FZD1 were significantly increased in high phosphorus group,and cell proliferation number was significantly decreased (P<0.01). Confocal laser microscopy showed that β-catenin was mostly expressed in the nucleus. Compared with the high phosphorus group,the apoptosis number,Ca2+ content,α-SMA,β-catenin,ALP,OPN,BMP2 and FZD1 expression of Danzhi group were significantly decreased,and the number of cell proliferation was significantly increased (P<0.01). Microscopic observation showed that the expression of β-catenin in the nucleus was decreased. Compared with DanZhi group,the apoptosis number,Ca2+ content,α-SMA,β-catenin,ALP,OPN,BMP2 and FZD1 expression were increased in LiCL group,and the number of cell proliferation was significantly decreased (P<0.01). Microscopic observation showed that the expression of β-catenin in the nucleus was increased.[Conclusion] Danzhi Jiangtang Capsule can reduce the calcification of lower limb endothelial cells in diabetic mice by down-regulating the expression of β-catenin in the nucleus,reducing the synthesis of β-catenin protein,inhibiting the transcription of phenotypic genes in downstream cells,and preventing and controlling vascular calcification of endothelial cells. |
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