China Animal Husbandry and Veterinary Medicine ›› 2023, Vol. 50 ›› Issue (6): 2245-2254.doi: 10.16431/j.cnki.1671-7236.2023.06.008

• Physiological and Biochemical • Previous Articles     Next Articles

Effect of Hypoxia on Proliferation and Oxidative Stress Injury of Pulmonary Artery Smooth Muscle Cells in Yaks

LI Rui1,2,3, ZHANG Yiyang1,2,3, LUO Xuehui1, ZHOU Manlin1,2,3, WU Shaomi1, YANG Kun1,2,3, QIAO Zilin1,2,3   

  1. 1. Life Science and Engineering College, Northwest Minzu University, Lanzhou 730030, China;
    2. Gansu Technology Innovation Center of Animal Cell, Biomedical Research Center, Northwest Minzu University, Lanzhou 730030, China;
    3. Key Laboratory of Biotechnology and Bioengineering of State Ethnic Affairs Commission, Lanzhou 730030, China
  • Received:2022-10-19 Online:2023-06-05 Published:2023-05-30

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Abstract: 【Objective】 This experiment was aimed to study the effects of hypoxia on the proliferation and oxidative stress injury of pulmonary artery smooth muscle cells (PASMCs) in yaks(Bos grunniens), and preliminarily explore the adaptive characteristics of pulmonary artery smooth muscle in yaks to hypoxia.【Method】 PASMCs of yaks were isolated and purified using α-smooth muscle actin (α-SMA).The test was divided into normoxia and hypoxia groups, and the proliferation efficiency of PASMCs in yaks was measured by CCK-8 method at 0, 2, 4, 6, 12, 24, 48, 72, 96 and 120 h under hypoxia and normoxia conditions.The protein concentrations of lactate dehydrogenase (LDH), superoxide dismutase (SOD), creatine kinase (CK) and malondialdehyde (MDA) in the cell culture supernatants of normoxia and hypoxia groups at 24, 48, 72 and 96 h of culture were measured by ELISA method.【Result】 The result of cell identification showed that more than 95% of cells could express α-SMA, which indicated that the isolated and purified cells were PASMCs in yaks.CCK-8 results showed that the cell proliferation folds in hypoxia group at 72, 96 and 120 h were extremely significantly lower than that in normoxia group (P<0.01), and the cell proliferation folds in both hypoxia and normoxia groups at 72 h were extremely significantly higher than 48 h (P<0.01), which indicated that the proliferation peak of PASMCs in yaks occurred at 72 h, and the prolonged hypoxia environment could reduce the proliferation efficiency of PASMCs in yaks.ELISA assay results were showed that compared with normoxia group, the LDH protein concentration of PASMCs culture supernatant of yaks in hypoxia group at 72 h was extremely significantly upregulated (P<0.01), the SOD protein concentration at 72 and 96 h were extremely significantly or significantly upregulated (P<0.01 or P<0.05), the CK protein concentration at 48 and 72 h were significantly or extremely significantly upregulated (P<0.05 or P<0.01), and the MDA protein concentrations at 72 h was significantly upregulated and at 96 h was significantly downregulated (P<0.05).Under hypoxia conditions, there was no significant difference of LDH and SOD protein concentrations in supernatant at different time (P>0.05), the CK and MDA protein concentrations at 24, 48 and 72 h were significantly higher than 96 h (P<0.05).Under normoxia conditions, the LDH protein concentration in supernatant at 48 h was significantly higher than 24 and 72 h (P<0.05), the SOD protein concentration at 24 and 48 h were significantly higher than 72 and 96 h (P<0.05), the CK protein concentration at 24 h was significantly higher than 72 and 96 h (P<0.05), the MDA protein concentration at 48 h was significantly higher than 72 h (P<0.05).【Conclusion】 Prolonged hypoxia could enhance oxidative stress and cell damage of PASMCs in yaks, weaken antioxidant effects, and significantly reduced cell proliferation efficiency.Four proteins of LDH, SOD, CK and MDA were played important regulatory roles in damage and anti-damage as well as antioxidant effects of PASMCs in yaks under hypoxia conditions, which might be related to the hypoxia adaptation of pulmonary vasculature in yaks.

Key words: yaks; pulmonary arterial smooth muscle cells (PASMCs); hypoxia; oxidative stress

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