四逆散对创伤后应激障碍大鼠海马组织分子表达的作用

Function of Sinisan on hippocampal molecular expression in rats with post-traumatic stress disorder

目的研究四逆散干预创伤后应激障碍大鼠的分子生物学机制。方法将SD大鼠50只按照体重平均分为5组:空白对照组、模型组、阴性对照组、阳性对照组、实验组。除了空白对照组以外,其他4组动物均制备模型。空白对照组不接受治疗,正常饲养。模型组不接受治疗。阴性对照组灌胃等体积0.9%Na Cl。阳性对照组灌胃盐酸帕罗西汀溶液0.42 mg·m L^(-1)。实验组灌胃四逆散水煎液(含生药0.24 g·m L^(-1))。于应激造模前1 h灌胃给药10 m L·kg^(-1)。每天灌服1次,共计7 d。于末次给药5 h后,各组统一采集海马组织。用HPLC法分析大鼠后海马组织匀浆的色谱峰。色谱条件:Agilent HC-C18(4.6 mm×250 mm,5μm);流速:0.8 m L·min^(-1);流动相A为甲醇,B为乙腈,C为0.05%磷酸水,梯度洗脱;检测波长:260 nm;柱温:23℃。结果与空白对照组1号、5号的峰面积(220.40±9.25),(86.27±7.39)m AU×min比较,模型组1号峰面积(82.50±9.60)m AU×min明显降低而5号峰面积(123±9.28)m AU×min明显升高,差异有统计学意义(均P<0.05)。与模型组比较,阴性对照组的1号峰面积(95.50±9.89)m AU×min、5号峰面积(119.30±9.93)m AU×min的色谱峰差异无统计学意义(P>0.05)。与阴性对照组比较,阳性对照组和实验组的1号峰面积(230.42±11.21),(234.80±12.47)m AU×min明显升高而5号峰面积(71.29±1.82),(72.16±2.55)m AU×min明显降低,差异有统计学意义(均P<0.05)。与阳性对照组比较,实验组新出现了8号、9号峰,提示这2个峰可能是四逆散内在化学成分或在机体内的代谢产物。结论四逆散对大鼠海马组织分子表达有明显的正向调节作用。

Objective To study the function mechanism of the Sinisanon on hippocampal molecular expression in rats with post -traumatic stress disorder. Methods Fifty SD rats were divided equally into 5 groups： blank control group, model group, negative control group group, positive control group and experimental group. The control blank group did not copy the model, do not receive treatment, the normal feeding. Model group, repetitive post -traumatic stress disorder model was induced by current stimulation, but not treated. Negative control group,equal volume of 0. 9% NaCl. Positive control group （ paroxetine hydrochloride 0. 42 mg·mL^-1） and experimental group （Sinisan, Containing crude drug 0. 24 g·mL^-1）. It given to drugs one hour before the model establish- ment. The rats were administered with 10 mL·kg^-1, once a day, for a total of 7 d. At the end of 5 hours after the last administration, the hippocampus tissues were collected. Fingerprints were used to compare the difference peaks of hippocampus tissue in the rats with post - traumatic stress disorder. High performance liquid chromatography conditions： the gradient elution. The chromatograph condition： Agilent HC -C18 （4. 6 mm × 250 mm, 5 μm）, speed of flow 0. 8 mL·min^-1; flowing A is methanol, flowing B is acetonitrile, flowing C is 0. 05% phosphorus acid water; wavelength of detection 260 nm; column warm 23℃. Results Compared with the blank control group, the model group is increased by 2nd, 3rd, 4th, 6th and 7th chromatographic peak. Compared with the blank control group on peak area of lth （220. 40 ±9.25） mAU×min ,5 th（86.27 ±7.39） mAU×min, peak area in model group lth （ 82.50±9.60 ） mAU × min were significantly decreased （ P 〈 0. 05 ）, 5th peak area （ 123.07 ± 9.28 ） mAU× min was significantly higher （ P 〈 0.05 ）. Compared with model group, the peak area at the time of lth （95.50 ±9. 89） mAU ×min, 5th （119.30 ±9.93） mAU ×min in negative controlgroup, it did not change significantly from chromatographic peak （P 〉 0. 05 ）. Compared with negative control group, positive control group and experimental group disappear by 2nd, 3rd, 4th, 6th and 7th peak, and lth （230. 42 ± 11.21 ）, （234. 80 ± 12.47） mAU×min peak area was significantly higher （P 〈0. 05） while 5th （71.29 ± 1.82）, （72. 16 ±2.55 ） mAU×min peak area were significantly decreased （P 〈 0. 05 ）. Compared with positive control group, experimental group increased by 8th and 9th , it hint the two peaks may be chemical constituents or metabolites in the organism of Sinisan.Conclusion The Sinisan have significant positive effect on hippocampal molecular expression.