CAJ | 학술논문

目的：研究高脂饮食对大鼠脑微血管内皮细胞HIF-1α及Claudin-5表达的影响，初步探讨高脂毒性对大鼠脑微血管的损伤机制。方法（1）40只SD大鼠随机分为高脂组与正常对照组，每组20只，分别予以高脂饲料和普通饲料喂养8周。（2）测定各组大鼠基础、第4周、第8周时体重及代谢指标变化。（3）8周时处死各组大鼠，免疫组化检测各组大鼠脑微血管内皮细胞上HIF-1α及Claudin-5蛋白表达情况，伊文氏蓝（EB）染色检测血-脑屏障（BBB）的通透性。结果（1）高脂组大鼠体重由基础值（165.00±12.100）g 增加至8周时的（401.30±66.827）g；对照组大鼠体重由基础值（163.00±10.100）g增加至8周时的（321.10±18.300）g，两组比较差异有统计学意义（P＜0.05）。（2）高脂组大鼠空腹血总胆固醇（TC）由基础值（1.576±0.1389）mmol/L升高为8周时的（2.032±0.3650）mmol/L，甘油三酯（TG）由基础值（0.601±0.1172）mmol/L升高至8周时的（2.679±1.0876）mmol/L，且显著高于相应时间点正常对照组（P＜0.05）。（3）8周时高脂组大鼠脑皮质微血管内皮细胞上，HIF-1α表达强于正常对照组（P＜0.05），Claudin-5表达呈弱阳性，正常对照组大鼠Claudin-5呈强阳性表达（P＜0.05）。（4）8周时高脂组大鼠EB含量较正常对照组升高，差异有统计学意义（P＜0.05）。结论高脂可通过上调HIF-1α蛋白的表达水平使紧密连接蛋白Claudin-5的表达下降，导致微血管病变及增加BBB的通透性。
목적：연구고지음식대대서뇌미혈관내피세포HIF-1α급Claudin-5표체적영향，초보탐토고지독성대대서뇌미혈관적손상궤제。방법（1）40지SD대서수궤분위고지조여정상대조조，매조20지，분별여이고지사료화보통사료위양8주。（2）측정각조대서기출、제4주、제8주시체중급대사지표변화。（3）8주시처사각조대서，면역조화검측각조대서뇌미혈관내피세포상HIF-1α급Claudin-5단백표체정황，이문씨람（EB）염색검측혈-뇌병장（BBB）적통투성。결과（1）고지조대서체중유기출치（165.00±12.100）g 증가지8주시적（401.30±66.827）g；대조조대서체중유기출치（163.00±10.100）g증가지8주시적（321.10±18.300）g，량조비교차이유통계학의의（P＜0.05）。（2）고지조대서공복혈총담고순（TC）유기출치（1.576±0.1389）mmol/L승고위8주시적（2.032±0.3650）mmol/L，감유삼지（TG）유기출치（0.601±0.1172）mmol/L승고지8주시적（2.679±1.0876）mmol/L，차현저고우상응시간점정상대조조（P＜0.05）。（3）8주시고지조대서뇌피질미혈관내피세포상，HIF-1α표체강우정상대조조（P＜0.05），Claudin-5표체정약양성，정상대조조대서Claudin-5정강양성표체（P＜0.05）。（4）8주시고지조대서EB함량교정상대조조승고，차이유통계학의의（P＜0.05）。결론고지가통과상조HIF-1α단백적표체수평사긴밀련접단백Claudin-5적표체하강，도치미혈관병변급증가BBB적통투성。
Objective To investigate the effect of high-fat diet on HIF-1αand Claudin-5 expression in rat brain microvascular endothelial cells, preliminary study injury mechanism of fat toxicity on rat brain microvascular. Methods (1)40 SD rats were randomly divided into high-fat group (n=20) and normal food groups (n=20), respectively to be fed with high fat diet and regular food eight weeks. (2)The weight, TG and TC of rats in each group were detected in baseline, 4 weeks and 8 weeks . (3)The rats were killed after 8 weeks, the protein level of HIF-1α, and Claudin-5 in the brain microvascular endothelial cells were determined by immunohistochemistry staining. Evans blue staining detect blood-brain barrier permeability. Results (1)High-fat groups' weight increased from (165.00±12.100)g to (401.30±66.827)g, control groups' weight increased from (163.00±10.100)g to (321.10±18.300)g, the difference was statistically significant (P<0.05) on 8 weeks; (2)High-fat groups' fasting TC by the underlying value of (1.576±0.138 9)mmol/L increased to 8 weeks of (2.032±0.365 0)mmol/L, TG by the underlying value of (0.601±0.117 2)mmol/L increased to (2.679±1.087 6)mmol/L at 8 weeks, and significantly higher than the corresponding time points normal control group (P<0.05). (3)High-fat group rat brain cortex microvascular endothelial cells, HIF-1α expression was stronger than the normal control group after 8 weeks (P<0.05), Claudin-5 expression was weakly positive, normal control rats Claudin-5 expression was strongly positive (P<0.05). (4)EB content of high-fat group increased compared with normal control group, the difference was statistically significant (P<0.05) after 8 weeks. Conclusion Hyperlipidemia can increase HIF-1αprotein levels and lead to tight junction protein Claudin-5 expression decreased, resulting in microvascular disease and increasing BBB permeability.