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

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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。 The adjuvant-induced arthritis model corresponding to the parasitism efficacy was established and the relationship between the effect of the rheumatism and its distribution in the rat body and the relationship of regression were observed. Freund’s complete adjuvant (0.1 m L) was injected into the left hind paw and was negative after 8 days. The model group was given normal saline. The positive control group was given tripterygium glycoside suspension (10 mg · kg-1) The middle and low dose groups were given the decoction of 10 g · kg -1, 5 g · kg -1 and 2.5 g · kg -1 for 21 d. On the 9th day after modeling, the rats’ left ankle and left ankle were measured once every 7 days. On the 22nd day, some rats in the high dose group of Sangaris were taken out of the eyeball at different time points to take blood and sacrificed, The rats in the liver, spleen, lung, kidney, stomach, large intestine, small intestine and brain tissue were taken out of the eyeball 30 min after administration, and the left hind foot ankle joint was taken. The levels of interleukin 1β ), Tumor necrosis factor-α (TNF-α), and the content of rutin, fructoside and quercitrin in the tissues of high-dose group were measured by HPLC. Relevant pharmacokinetic parameters were analyzed by DAS 2.0. The results showed that Sanguisheng Decoction significantly improved the condition of foot swelling in adjuvant-induced arthritis rats and decreased the levels of IL-1β and TNF-α in serum. Rat efficacy substance analysis showed that only a small amount of rutin in the stomach distribution, the AUC0-t for the accumulation of glycosides in the stomach of stomach> small intestine> kidney, body maintenance time for the kidney = stomach> small intestine> lung> heart; quercetin Glycosides in the organization AUC0-t for the stomach> kidney> liver> heart> lung> spleen> small intestine> brain> large intestine> serum in vivo maintenance time for the kidney = liver = small intestine = brain = lung = spleen = heart = stomach> serum. It was concluded that the Sang-ssu decoction can be effective adjuvant arthritis model rats. Eupatorium glycosides, quercitrin Sang parasitic rheumatism effect of an important material. Eupatorium glycosides, quercitrin distribution in rats and Sang parasitic kidney, the traditional understanding of the liver.
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