岩黄连总碱在抑郁模型大鼠体内的药代动力学分析

Pharmacokinetics of Total Alkaloids of Corydalis saxicola in Depression Model Rats

目的:探讨岩黄连总碱对脂多糖(LPS)诱导抑郁大鼠模型的影响,以及其中8种主要成分的药代动力学特征。方法:将雄性24只SD大鼠随机分成正常组、模型组、氟西汀组(10 mg·kg^(-1))、岩黄连总碱组(210 mg·kg^(-1)),每组6只。除正常组外,其他大鼠经腹腔注射LPS建立抑郁症炎症模型,造模1周后开始给药,各给药组按相应剂量灌胃药液,正常组及模型组灌胃等体积蒸馏水,边造模边给药。连续给药2周后,通过糖水偏好、强迫游泳及旷场试验测试岩黄连总碱对抑郁大鼠行为学的影响,酶联免疫吸附测定法(ELISA)测定大鼠血清中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、IL-6水平,苏木素-伊红(HE)染色观察大鼠海马组织病理学变化。末次给药后,按设定时间经眼眶取血,采用超高效液相色谱-三重四极杆串联质谱法(UPLC-QqQ-MS)检测其中脱氢卡维丁、延胡索乙素、黄连碱、巴马汀、药根碱、小檗碱、小檗红碱、表小檗碱的血药浓度,绘制药-时曲线,通过DAS 2.0软件对各成分药代动学参数进行分析。结果:与正常组比较,模型组糖水偏好率降低、旷场总路程减少、游泳不动时间延长,血清中炎症因子表达升高(P<0.01);与模型组比较,各给药组大鼠糖水偏好率增加、旷场总路程增加,游泳不动时间缩短,岩黄连总碱组血清中炎症因子表达降低(P<0.05,P<0.01)。HE染色结果显示,模型组大鼠海马区神经元缺失、排列紊乱且有残留空泡,岩黄连总碱组大鼠海马区神经元增多且排列整齐、胞浆清楚。药代动力学结果显示,经连续灌胃岩黄连总碱后,8种活性成分的达峰时间(t_(max))为0.19~2.06 h,半衰期(t_(1/2))为3.71~8.70 h,其中延胡索乙素的药时曲线下面积(AUC_(0-∞))最高且t_(1/2)最短,黄连碱、巴马汀、药根碱、小檗碱、小檗红碱、表小檗碱的AUC_(0-∞)均较低。脱氢卡维丁、黄连碱、巴马汀、小檗碱和表小檗碱药时曲线出现明显的双峰现象。结论:岩黄连总碱能够改善大鼠抑郁样行为,抑制血清中炎症因子的表达,改善海马区病理损伤,具有抗抑郁作用;同时,该有效部位在抑郁模型大鼠体内吸收较快,消除较慢,药效维持时间长。

Objective:To investigate the effect of total alkaloids of Corydalis saxicola on a rat model of lipopolysaccharide(LPS)-induced depression,as well as the pharmacokinetic characteristics of 8 of its major components.Method:Twenty-four male SD rats were randomly divided into normal group,model group,fluoxetine group(10 mg·kg^(-1))and total alkaloids of C.saxicola group(210 mg·kg^(-1)),with 6 rats in each group.In addition to the normal group,the rats were injected intraperitoneally with LPS to establish the inflammation model of depression,and the drug administration was started 1 week after modeling,and the administration groups were gavaged according to the corresponding dose,and the normal and model groups were intragastric administration with equal volume of distilled water,and the administration was performed along with the modeling.After two weeks of continuous administration,the effect of total alkaloids of C.saxicola on the behavior of depressed rats were tested by sucrose preference,forced swimming and open field experiments,the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-1βand IL-6 in serum of rats were determined by enzyme-linked immunosorbent assay(ELISA),the histopathological changes of rat hippocampus were observed by hematoxylin-eosin(HE)staining.After the last administration,blood was collected from orbit according to the set time,and ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry(UPLC-QqQ-MS)was established to simultaneously detect the concentrations of dehydrocavidine,tetrahydropalmatine,coptisine,palmatine,jatrorrhizine,berberine,berberrubine and epiberberine in plasma,and drug-time curves were drawn.The pharmacokinetic parameters were analyzed by DAS 2.0 software.Result:Compared with the normal group,the model group exhibited a decrease in sucrose preference rate,total distance traveled in the open field,as well as an increase in swimming immobility time and serum inflammatory factor expression(P<0.01).In contrast,compared with the model group,rats in each administration group showed an increase in sucrose preference rate and total distance traveled in the open field,a decrease in swimming immobility time,and a reduction in serum inflammatory factor expression(P<0.05,P<0.01).Additionally,HE staining results revealed that neurons in the hippocampus of rats from the model group were characterized by loss,disorganization and residual vacuoles,whereas those from the total alkaloids of C.saxicola group displayed an increase in number with orderly arrangement and clear cytoplasm.Pharmacokinetic results showed that the time to peak(t_(max))and half-life(t_(1/2))of the 8 active ingredients were 0.19-2.06 h and 3.71-8.70 h after continuous administration of total alkaloids of C.saxicola.Among them,the area under the curve(AUC_(0-∞))of tetrahydropalmatine was the highest and the t_(1/2) was the shortest,and the AUC_(0-∞) of coptisine,palmatine,jatrorrhizine,berberine,berberrubine and epiberberine were low.The curves of dehydrocavidine,coptisine,palmatine,berberine and epiberberine showed obvious double peak phenomenon.Conclusion:Total alkaloids of C.saxicola can improve the depression-like behavior of rats,inhibit the expression of inflammatory factors in serum,improve the pathological injury of hippocampus,and has the antidepressant effect.Meanwhile,the effective site is absorbed quickly and eliminated slowly in the depressed model rats,and the efficacy is maintained for a long time.