2023, Vol. 27 ›› Issue (26): 4132-4138

Protective effect of ethanol extract of Zizhu on vascular endothelial cell injury induced by high glucose and high lipids

Wang Lijuan1, 2, Liu Guobin2, Han Qiuqin1, Zhao Zhihong1, He Jinjing1, 2, Li Wenhui1, 2   

Abstract: BACKGROUND: Zizhu ointment is composed of Astragali Radix, Radix Arnebiae seu Lithospermi, Cinnabar, Resina Dracoins, Ejiao and Borneol, which has the effects of clearing heat and detoxifying, removing saprophyxia and strengthening muscles, invigorating qi and benefiting blood. Previous clinical and animal experiments have shown that it can inhibit inflammation and promote angiogenesis in the healing process of diabetic ulcer.
OBJECTIVE: To investigate the protective effect of ethanol extract of Zizhu on the injury of rat brain microvascular endothelial cells (bEend.3 cells) induced by high glucose and high lipids at the cellular level, and to study its effect on angiogenesis.
METHODS: bEnd.3 cells were induced with glucose and palmitic acid at different concentrations, and treated with different concentrations of ethanol extract of Zizhu. The optimum concentrations for modeling and administration were identified by cell counting kit-8 method. The effect of ethanol extract of Zizhu on angiogenesis of bEnd.3 cells was observed by scratch test and angiogenesis test. AnnexinV/PI apoptosis detection kit was used to detect the apoptosis rate of bEnd.3 cells under fluorescence microscope. The protein and mRNA expression levels of VEGFA, Ang-2, SPRED1, and PIK3R2 in bEnd.3 cells were determined by western blot and qRT-PCR, respectively.
RESULTS AND CONCLUSION: Cell viability was detected by cell counting kit-8 method. 25 mmol/L glucose+200 μmol/L palmitic acid for 24 hours was selected as the model group; treatment with ethanol extract of Zizhu 500 mg/L for 24 hours was performed as the way of administration in the ethanol extract of Zizhu group. Compared with the normal group, bEnd.3 cell migration and angiogenesis were decreased in the model group (P < 0.001, P < 0.05), bEnd.3 cell apoptosis rate was increased (P < 0.001), the mRNA and protein expressions of VEGFA and Ang-2 were significantly decreased (P < 0.001), and the mRNA and protein expressions of SPRED1 and PIK3R2 were significantly increased (P < 0.001). Compared with the model group, in the ethanol extract of Zizhu group, cell migration and angiogenesis of bEnd.3 cells were significantly increased (P < 0.05), bEnd.3 cell apoptosis rate was significantly decreased (P < 0.001), mRNA and protein expressions of VEGFA and Ang-2 were significantly up-regulated (P < 0.001), mRNA and protein expressions of SPRED1 and PIK3R2 were significantly down-regulated (P < 0.001, P < 0.05). All the experimental results show that the ethanol extract of Zizhu can protect bEnd.3 cells induced by high glucose and high lipid, promote cell migration and angiogenesis, and reduce cell apoptosis.

Key words: ethanol extract of Zizhu, high glucose and high lipid, mouse brain microvascular endothelial cell, angiogenesis, vascular endothelial growth factor