ABSTRACT
ObjectiveTo investigate the possible mechanisms of modified Xiaoyao Powder （逍遥散） in the treatment of hyperprolactinemia （HPRL） with liver constraint and spleen deficiency. MethodsNinety-six female SD rats were randomly divided into a normal group （n=16） and a modeling group （n=80）. In the modeling group， rats were subjected to chronic unpredictable stress combined with intraperitoneal injection of metoclopramide to establish a rat model of HPRL with liver constraint and spleen deficiency. The 80 successfully modeled rats were randomly divided into a model group， a high， medium， and low-dose group of modified Xiaoyao Powder， and a bromocriptine group， with 16 rats in each group. The high， medium， and low-dose groups of modified Xiaoyao Powder were orally administered doses of 60， 30， and 15 g/（kg·d） respectively， the bromocriptine group was orally administered bromocriptine tablets at a dose of 1 mg/（kg·d）， and the normal group and model group were orally administered 10 ml/（kg·d） of normal saline for 14 consecutive days. ELISA was used to detect serum prolactin （PRL） level； immunohistochemistry was used to determine the expression of tumor necrosis factor-alpha （TNF-α） and tyrosine hydroxylase （TH） in the hypothalamus； Western blot was used to detect the protein expression of tumor necrosis factor receptor 1 （TNFR1） in the hypothalamus； Real-time PCR was used to detect the mRNA expression of receptor interacting protein kinase 3 （RIP3） in the hypothalamus； immunofluorescence was used to detect the co-expression of RIP3 and dopamine neurons in the hypothalamus. ResultsCompared with the normal group， the serum PRL levels were increased in the model group， and the expression of hypothalamic TNF-α， TNFR1， RIP3 mRNA， and the co-expression of RIP3 with dopamine neurons were significantly increased， while TH expression was decreased （P＜0.05 or P＜0.01）. Compared with the model group， the expression of hypothalamic TNF-α was decreased in the bromocriptine group and low-dose group of modified Xiaoyao Powder， and the expression of TH was significantly increased in the medium and high-dose groups of modified Xiaoyao Powder and the bromocriptine group. The serum PRL levels， hypothalamic TNFR1 and RIP3 mRNA expression， and the co-expression of RIP3 with dopamine neurons were significantly decreased in all dose groups of modified Xiaoyao Powder and the bromocriptine group （P＜0.05 or P＜0.01）. Compared with the bromocriptine group， the serum PRL level were significantly increased in the high and low-dose groups of modified Xiaoyao Powder， TH expression was significantly increased in the medium-dose group of modified Xiaoyao Powder， hypothalamic RIP3 mRNA expression was decreased in the low-dose group of modified Xiaoyao Powder， and the co-expression of RIP3 with dopamine neurons was significantly increased in the high-dose group of modified Xiaoyao Powder （P＜0.01）. ConclusionModified Xiaoyao Powder can regulate the programmed cell death of hypothalamic dopamine neurons， affect DA expression， and regulate PRL levels， which may be one of its mechanisms in the treatment of HPRL with liver constraint and spleen deficiency.
ABSTRACT
Objective To establish a quality traceability evaluation method for the whole honeysuckle oral solution process by identifying and screening its anti-inflammatory quality markers.Methods UPLC/-TOF-MS was used to analyze the iridoids and phenolic acids in oral solution,and the correlations were constructed by molecular network technology.The HPLC fingerprints of multiple batches of oral solution were established,and similarity analyses were performed to identify key pharmacodynamic molecules.The key anti-inflammatory quality markers were confirmed by the NF-κB dual luciferase assay system.Further,the quantification of 12 quality markers of iridoids and phenolic acids in oral solution was established separately based on the dual-wavelength HPLC technique.The quality of the oral solution was evaluated by examining the extraction and transfer rate of quality markers during the processing of raw materials and preparations and thermal stability.Results A total of 9 iridoids and 6 phenolic acids were identified in the oral solution,and the possible conversion relationships between their components were depicted.Fingerprint analysis of 11 batches of oral liquids showed that the composition of their main peaks was the same,with a similarity of more than 90%.Among them,6 iridoids(loganic acid,secologanoside,secologanic acid,sweroside,secoxyloganin,secologanin)and 6 phenolic acids(neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,isochlorogenic B,isochlorogenic A,isochlorogenic C)exhibited NF-κB inhibitory activity,which were the main pharmacological components and could be used as quality markers.The traceability of the above 12 quality markers was investigated in a multi-batch process based on the dual-wavelength HPLC method.The thermal stability studies of the raw materials revealed that the contents of their total iridoids and phenolic acids remained stable.Still,some of them would be transformed between components.The production process of the oral solution was stable,and the transfer rates of the iridoids and phenolic acids during the extraction,concentration and preparation were over 76%and 63%,respectively.Conclusion The method is stable,reliable,easy to operate and can evaluate the full honeysuckle oral solution process,which provides an effective means for the quality control of honeysuckle herbs and preparations.
ABSTRACT
@#Objective To investigate the effect of Ski on the secretion of inflammatory cytokines from activated astrocytes. Methods Astrocytes were obtained from cerebral cortex of a three-day old Sprague-Dawley rat and cultured in vitro. They were divided into blank group, control group and siRNA group. The Ski gene was silenced in siRNA group. The expression of Ski was tested with Western blotting and immunofluorescence 48 hours later. Then the astrocytes were stimulated with lipopolysaccharide for 24 hours. The secretion of tumor necrosis factor α (TNF-α) and interleukin-1β (IL-1β) in activated astrocytes was detected with ELISA. Results The expression of Ski protein reduced in the siRNA group (P<0.001), as well as the secretion of TNF-α and IL-1β (P<0.001). Conclusion Ski may play a role in inflammatory response of astrocyte.