急性高原缺氧大鼠脑皮质相关细胞因子变化及蕨麻的保护作用

Changes in cytokines related to cerebral cortex with acute high altitude hypoxia in rats and protective effects of Potentilla anserina L.

目的探讨模拟高原缺氧条件下大鼠脑皮质内细胞因子变化及蕨麻的保护作用。方法选择成年健康雄性SD大鼠54只，按随机数字表法分为平原组、模拟中海拔（3000m）组和模拟高海拔（7000m）组，后两组再分为缺氧12h和24h两个时间点，各时间点又分蕨麻组和对照组，每组6只。建立大鼠急性低压缺氧损伤模型；蕨麻组灌胃蕨麻提取物300mg／kg，对照组给予生理盐水。于各时间点处死大鼠，剥离脑组织，匀浆离心后取上清，采用悬浮芯片蛋白法测定单核细胞趋化蛋白-1（MCP-1）、白细胞介素（IL-113、-6、-10）、肿瘤坏死因子-α（TNF-α）、γ-干扰素（IFN-γ）及血管内皮生长因子（VEGF）含量。结果与平原组比较，模拟中、高海拔对照组各时间点MCP-1（ng／L）、IL-1β（ng／L）、IL-6（ng／L）、IL-10（ng／L）、TNF-α（ng／L）、IFN-γ（ng／L）及VEGF（ng／L）含量均显著升高，且上述各指标均随时间延长和海拔升高而升高，以模拟高海拔对照组24h升高最显著（MCP-1：199．69±16．67比164．32±11．84；IL-1β：83．32±9．90比17．05±5．60；IL-6：112．15±4．55比74．50±10．00；IL-10：129．49±16．28比68．11±10．26；TNF—α：106．29±13．05比54．54±6．97；IFN-γ：105．72±14．76比51．77±11．59；VEGF：227．95±26．84比80．28±23．76，均P〈0．01）。与同海拔对照组同期比较，模拟中、高海拔蕨麻组各时间点MCP-1、IL-1β、TNF—α、IFN-γ含量均明显降低（高海拔蕨麻24h组MCP-1：176．48±16．23比199．69±16．67；IL-1β：67．66±12．34比83．32±9．90；TNF-α：82．26±8．26比106．29±13．05；IFN-γ：83．29±9．84比105．72±14．76，均P〈0．05），IL-6、IL-10及VEGF含量均显著升高（高海拔蕨麻24h组IL-6：122．40±6．35比112．15±4．55；IL-10：150．66±11．18比129．49±16．28；VEGF：280．98±22．72比227．95±26．84，均P〈0．05）。结论高原急性缺氧使炎症细胞因子生成增加；蕨麻可通过降低促炎症细胞因子含量，升高抗炎症细胞因子含量来减少脑组织氧化应激损伤。

Objective To explore the changes in levels of cytokines in rats cerebral cortex under simulate high altitude hypoxia and to analyze the protective effects of Potentilla anserina L. on the above indexes. Methods Fifty- four healthy male Sprague Dawley （SD） rats were randomly divided into plain group, simulated intermediate altitude （3000 m） group and simulated high altitude （7000 m） group. Except plain group, all rats in hypobaric hypoxia injury groups were subdivided into groups of hypobarie hypoxia 12 hours and 24 hours （time points） according to the duration of hypobaric hypoxia exposure, and the rats at 12 hours and 24 hours were further subdivided into control group and Potentilla anserina L. group, with 6 rats in each group. The rat model of acute hypobaric hypoxia injury was reproduced. Rats in the Potentilla anserina L. group and control group were administered with Potentilla anserina L. （300 mg/kg）and normal saline, respectively into the stomach. Rats were sacrificed at the above time points, and the brain tissue was harvested, homogenized and centrifugalized, then the supernatant was collected to determine the levels of monocyte chemoattractant protein-1 （MCP-1）, intedeukins （IL-1 β, -6, -10）, tumor necrosis factor- α （TN F- α ）, γ-interferon （ IFN- γ） and vascular endothelial growth factor （VEGF） by suspension array technique. Results Compared with plain group, the levels of MCP-1 （ng/L）, IL-1β （ng/L）, IFN-γ （ng/L）, IL-6 （ng/L）, IL-10 （ng/L）, VEGF （ng/L） and TNF-α（ng/L） raised significantly in every time point of simulated intermediate and high altitude groups, and along with the prolongation of time and the elevation of the altitude, the above indexes were increased with the increase in high altitude 24 hours control group the most significant （MCP-1 ： 199.69±16.67 vs. 164.32±11.84 ; IL-1β ： 83.32±9.90 vs. 17.05±5.60 ; IL-6 ： 112.15±4.55 vs. 74.50±10.00 ; IL-10 ： 129.49±16.28 vs. 68.11±10.26 ; IFN-β ： 105.72±14.76 vs. 51.77±11.59, TNF-α ： 106.29±13.05 vs. 54.54-±6.97 ; VEGF ： 227.95-± 26.84 vs. 80.28± 23.76, all P〈0.01 ）. Compared with the same altitude control group, the simulated intermediate and high altitude treatment groups at the time points, the levels of MCP-1, IL-1β, IFN-γ and TNF-α were significantly decreased （high altitude 24 hours Potentilla anserina L. group MCP-1 ： 176.48± 16.23 vs. 199.69±16.67; IL-113：67.66±12.34 vs. 83.32±9.90; TNF-α ：82.26±8.26 vs. 106.29±13.05; IFN-γ ： 83.29±9.84 vs. 105.72± 14.76, all P〈0.05）, while the levels of IL-6, IL-10 and VEGF were obviously increased （high altitude 24 hours Potentilla anserina L. group IL-6 ： 122.40±6.35 vs. 112.15±4.55 ; IL-10 ： 150.66± 11.18 vs. 129.49±16.28 ; VEGF ： 280.98±22.72 vs. 227.95±26.84, allP〈0.05）. Conclusion Simulated acute high altitude exposure may induce hypobaric hypoxia injury in rats, resulting in increase of the production of cytokines, and Potentilla anserina L. can significantly decrease the levels of pro-inflammatory cytokines and increase the levels of anti-inflammatory eytokines, thus it can protect the cerebral cortex against the damage induced by hypoxia.