摘要
目的探究不同浓度1,25-二羟维生素D3[1,25-(OH)_(2)D_(3)]对高原肺水肿(High altitude pulmonary edema,HAPE)雄性大鼠的防治作用及可能机制。方法将105只雄性SD大鼠随机分为对照组、低氧组和5个不同浓度组(低氧+浓度A-E组)。不同浓度组大鼠每天灌胃[应用1,25-(OH)_(2)D_(3)]一次,5天后,低氧组和5个浓度组均置于模拟海拔6000 m的低压舱48 h,检测大鼠血气指标氧饱和度(Oxygen Saturation,SaO_(2))、氧分压(Partial pressure of oxygen,PaO_(2))及肺动脉压(Pulmonary artery pressure,PAP)后处死大鼠,测定肺组织干湿重比,通过HE染色法观察大鼠肺组织病理学变化,用ELISA法检测超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)及丙二醛(MDA)活性含量,用RT-PCR法测定大鼠肺组织中炎症因子肿瘤坏死因子α(TNF-α)、白细胞色素1β(IL-1β)、白细胞色素6(IL-6)和CYP27B1、VDR mRNA水平,用蛋白质免疫印迹实验(Western Blot)法检测CYP27B1、VDR蛋白水平。结果1,25-(OH)_(2)D_(3)降低HAPE大鼠PAP、LWC,提高SaO_(2)及PaO_(2),降低肺组织IL-1β、IL-6、TNF-α水平(P<0.05),升高SOD、GSH-Px酶活性(P<0.05),降低MDA含量,上调VDR蛋白水平,下调CYP27B1水平,可减轻肺水肿程度。预防性给予1,25-(OH)_(2)D_(3)(浓度0.252μg/kg)能够有效预防HAPE的发生。结论初步明确了浓度为0.252μg/kg(浓度D组)的1,25-(OH)_(2)D_(3)对雄性大鼠有较好的防治HAPE的效果;这种预防及保护作用可能与1,25-(OH)_(2)D_(3)下调CYP27B1、上调VDR表达有关。
Objective To explore the prevention and treatment effects of different concentrations of 1,25-dihydroxyvitamin D_(3)[1,25-(OH)_(2)D_(3)]on high altitude pulmonary edema(HAPE)and its possible mechanism.Methods 105 male SD rats were randomly divided into control group,hypoxia group and 5 different concentrations of 1,25-dihydroxyvitamin D_(3) groups(hypoxia+concentration A-E group).The rats in different concentration groups were given intragastric administration[application of 1,25-(OH)_(2)D_(3)]once a day.Both the hypoxia group and 5 concentration groups were placed in a low pressure chamber at a simulated altitude of 6000 m for 48 h 5 days later.The rats were sacrificed after Oxygen Saturation(SaO_(2)),Partial pressure of oxygen(PaO_(2))and Pulmonary artery pressure(PAP)were detected.The dry-wet weight ratio(LWC)of lung tissue was determined.The histopathological changes of lung were observed by HE staining.The activity contents of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)were detected by ELISA.The mRNA levels of inflammatory cytokines such as tumor necrosis factor-α(TNF-α),leukin 1β(IL-1β),leukin 6(IL-6),and CYP27B1 or VDR in rat′s lung tissues were determined by RT-PCR.The protein levels of CYP27B1 and VDR were detected by Western Blot.Results 1,25-(OH)_(2)D_(3) decreased the levels of PAP and LWC,and increased the levels of SaO_(2) and PaO_(2).It decreased IL-1β,IL-6 and TNF-αlevels in lung tissue(P<0.05),and increased the enzyme activities of SOD and GSH-Px(P<0.05).1,25-(OH)_(2)D_(3) decreased MDA content and up-regulated VDR protein level in HAPE rats.Moreover,down-regulating the level of CYP27B1 could reduce the degree of pulmonary edema.Prophylactic administration of 1,25-(OH)_(2)D_(3)(0.252μg/kg)could effectively prevent HAPE.Conclusion It is preliminary clear that 1,25-(OH)_(2)D_(3) with concentration of 0.252μg/kg(concentration D group)has a better effect on the prevention and treatment of HAPE.The preventive and protective effects may be related to the d
作者
李天天
张先钧
戴重阳
朱梦婷
邓章昌
蒲小燕
Li Tiantian;Zhang Xianjun;Dai Chongyang;Zhu Mengting;Deng Zhangchang;Pu Xiaoyan(Medical Department of Qinghai University,Xining 810001,China)
出处
《中国高原医学与生物学杂志》
CAS
2023年第4期255-262,共8页
Journal of Chinese High Altitude Medicine & Biology
基金
国家自然科学基金项目(82160322)
青海省应用基础项目(2021-SF-153)。
