基于“病证-效应-生物样本分析”方法的桑寄生祛风湿功效物质及归经研究

Antirheumatic substance and meridian tropism of Loranthus parasiticus based on "syndrome-efficacy-analysis of biological samples"

实验建立与桑寄生功效相对应的佐剂型关节炎模型,观察其祛风湿功效物质及其在大鼠体内分布与归经的关系。通过左后足注射弗氏完全佐剂(0.1 m L),8 d后阴性、模型组给予生理盐水,阳性对照组给予雷公藤多苷混悬液(10 mg·kg-1),桑寄生高、中、低剂量组给予桑寄生高(10 g·kg-1)、中(5 g·kg-1)、低(2.5 g·kg-1)剂量水煎液,连续给药21 d。造模第9天起,每7 d测1次大鼠左后足踝关节直径,给药第22天桑寄生高剂量组部分大鼠于不同时间点摘眼球取血,处死,并取心、肝、脾、肺、肾、胃、大肠、小肠、脑组织,其余大鼠均在给药30 min后摘眼球取血,取左后足踝关节,ELISA法检测血清中白介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)含量,HPLC检测高剂量组大鼠各组织中芦丁、萹蓄苷、槲皮苷的含量,DAS 2.0分析相关药动学参数。结果表明,桑寄生水煎液能明显改善佐剂型关节炎模型大鼠足肿胀状况,且能降低大鼠血清中IL-1β,TNF-α含量。大鼠体内功效物质分析结果显示芦丁仅在胃中有少量分布,萹蓄苷在组织中AUC0-t为胃>小肠>肾,体内维持时间为肾=胃>小肠>肺>心;槲皮苷在组织中AUC0-t为胃>肾>肝>心>肺>脾>小肠>脑>大肠>血清,体内维持时间为肾=肝=小肠=脑=肺=脾=心=胃>大肠>血清。由此得出,桑寄生水煎液可有效治疗佐剂关节炎模型大鼠。萹蓄苷、槲皮苷为桑寄生祛风湿功效的重要物质。萹蓄苷、槲皮苷在大鼠体内分布与桑寄生归肾、肝经的传统认识一致。

To study the antirheumatic substance of Loranthus parasiticus and observe the relationship between its in vivo distribution and meridian tropism in rats by establishing adjuvant arthritis models corresponding to effectiveness. All rats except the negative control group were injected with 0. 1 m L Freund＇s complete adjuvant on the left foot. After 8 days,the rats in negative control group and model group were given with normal saline while the rats in positive control group were given with tripterygium glycosides suspension 10 mg·kg^- 1,and the rats in L. parasiticus treatment groups were given with high（ 10 g·kg^- 1）,medium（ 5 g·kg^- 1） and low（ 2. 5 g·kg^- 1） dose decoction for 21 days. The left rear ankle joint diameter of rats were measured every 7 days from the 9th day of modeling.On the 22 nd day,eyeball blood of part rats in L. parasiticus high-dose group was taken at different time points,and then they were sacrificed to take heart,liver,spleen,lung,kidney,stomach,large intestine,small intestine and brain tissues. For the remaining rats,eyeball blood was taken 30 min after drug treatment,and their left rear ankle joints were taken to detect interleukin（ IL）-1β and tumor necrosis factor（ TNF）-α levels in serum by ELISA method; rutin,avicularin and quercitrin levels in the tissues of high-dose group were detected by HPLC; pharmacokinetic parameters were analyzed by using DAS 2. 0. Our results showed that L. parasiticus decoction could significantly improve the paw edema situation of adjuvant arthritis model rats,and reduce IL-1β and TNF-α levels in rat serum.The in vivo efficacy substance analysis in rats showed that rutin was only present in the stomach with a small amount. AUC0-tof avicularin was stomach 〉small intestine〉 kidney,and the duration time in vivo was kidney = stomach〉 small intestine〉 lung〉 heart. AUC0-t of quercitrin was stomach 〉kidney 〉liver heart〉 lung 〉spleen 〉small intestine〉 brain〉 large intestine〉 serum,and the duration time in vivo was kidney = liver = small intestine = brain = lung = spleen = heart = stomach 〉large intestine〉 serum. The research indicated that L. parasiticus decoction was effective in treating rats with adjuvant arthritis. Avicularin and quercitrin are important ingredients of L. parasiticus in antirheumatism therapy. The distribution of avicularin and quercitrin in rats were consistent with traditional understanding that L. parasiticus could attribute to the kidney and liver meridians.