沙棘果油对慢性疲劳综合征大鼠NK细胞杀伤活性的影响及机制

Effects of Sea Buckthorn Fruit Oil on Killing Activity of Natural Killer Cells in Rats with Chronic Fatigue Syndrome(CFS)and the Mechanism

目的:研究沙棘果油对慢性疲劳综合征(chronic fatigue syndrome,CFS)大鼠NK细胞杀伤活性的影响及机制。方法:采用游泳结合束缚制备CFS动物模型,以沙棘果油1 m L灌胃干预,给药21 d后检测指标,采用免疫磁珠分离大鼠脾脏NK细胞,采用LDH法检测NK细胞杀伤活性,流式细胞术检测NK细胞在全血中的分布,realtime PCR检测NK细胞穿孔素、颗粒酶、IFN-γm RNA表达水平。结果:与正常对照组相比,CFS模型组大鼠NK细胞杀伤活性、全血NK细胞数量、NK细胞穿孔素、颗粒酶B及IFN-γm RNA表达水平均明显降低(P<0.05)。与CFS模型组相比,CFS+沙棘果油干预组大鼠NK细胞杀伤活性、全血NK细胞比例明显升高(P<0.05),并且此两项指标与正常对照组相比无明显差异(P>0.05)。与CFS模型组大鼠相比,CFS+沙棘果油干预组大鼠穿孔素、颗粒酶B m RNA表达均明显上调(P<0.05),穿孔素m RNA表达水平与正常对照组大鼠相比无明显差异(P>0.05)。结论:沙棘果油可通过提高NK细胞在全血中的数量、促进穿孔素和颗粒酶的表达提高CFS大鼠NK细胞杀伤活性。

Objective：To explore the effects of sea buckthorn fruit oil on killing activity of natural killer cells（NK）cells in rats with CFS and the mechanism. Methods：Swimming combined with constraint was used to establish CFS animal models. The rats were given 1 m L of sea buckthorn fruit oil through lavage,and were detected for indicators after 21 days of medication. Immune magnetic bead was used to separate NK cells from rat spleen. LDH method was used to detect killing activity of NK cells. Flow cytometry was used to detect the distribution of NK cells in whole blood. And real-time PCR was used to detect the expressions of perforin,granular enzyme and IFN-γ m RNA in NK cells. Results：Compared with the normal control group,CFS model group was significantly lower in killing activity of NK cells,number of NK cells in whole blood and expressions of perforin,granular enzyme B and IFN-γ m RNA in NK cells（P〈0.05）. Compared to CFS model group,CFS ＋ sea buckthorn fruit oil intervention group was obviously higher in killing activity of NK cells and proportion of NK cells in whole blood（P〈0.05）,and these two indicators were not significantly different compared to the normal control group（P〈0.05）. Compared to CFS model group,CFS ＋ sea buckthorn fruit oil intervention group was obviously higher in expression of perforin and m RNA of granzyme B（P〈0.05）,and the expressions of perforin m RNA were not significantly different compared to the normal control group（P〈0.05）. Conclusion：Sea buckthorn fruit oil can improve the killing activity of NK cells in CFS rats by increasing the number of NK cells in whole blood and expressions of perforin and granzyme.