清热功效指引下的新疆软紫草活性分子筛选与解热镇静作用评价

Screening of active molecules and evaluation of antipyretic and sedative effects of Arnebiaeuchroma(Royle)Johnst.under the guidance of heat-clearing efficacy

目的新疆软紫草性寒味苦,主清热凉血,基于此功效本文从系统药理学角度虚拟筛选其成药性分子并评价代表化合物的解热镇静作用。方法基于TCMSP等数据库构建软紫草成分库,通过admet SAR法筛选成药性较佳的活性分子,基于bSDTNBI法预测靶标并构建网络关系,通过MCODE拓扑学分析关键靶点及通路,autoDOCK验证了紫草主要化学成分与作用靶点可能存在的机制。建立2,4-二硝基苯酚(DNP,25 mg·kg^(-1))诱导的血热证小鼠模型,测定小鼠体温、饮水量、尿液量、粪便含水量及凝血功能指标,行为学体系评价代表化合物的镇静作用,包括自主活动、体温、睡眠作用,旷场实验分析运动轨迹。结果根据ADME性质以OB 30%,DL>0.18及CYP450代谢酶综合打分筛选出排名前20的活性分子,主要活性代表分子为紫草素(SK),涉及神经配体-受体关系和癌症等通路,MCODE分析显示紫草与OPRK1,GABRG2等中枢神经靶点密切相关,进一步药靶互作指出SK可与GABRG2等稳定结合。体内药效学结果表明SK组小鼠饮水量减少、尿液量和粪便含水量增加;SK显著降低了小鼠的自主活动次数及体温且具有量效一致性,缩短了睡眠潜伏期,显著延长了戊巴比妥钠诱导的睡眠时间(P<0.01);旷场实验指出SK显著降低了DNP小鼠的运动总距离和平均速度,并显示了其运动轨迹的减少(P<0.01)。结论清热功效指引下的新疆软紫草成药性分子以萘醌类紫草素为主,其具有一定的中枢性解热与部分中枢抑制作用,具体表现在SK可有效抑制血热引起的中枢兴奋,且这一作用与γ-氨基丁酸受体(GABA)突触后抑制及阿片受体激活有关。

Objective Arnebiaeuchroma(Royle)Johnst.is bitter and cold,its main effect is to clear heat and cool blood.Based on this effect,the pharmacological effect of antipyretic and sedative on the central nervous system is unknown.In this paper,from the perspective of systemic pharmacology,virtual screening of its druggable molecules and evaluation of the central role of representative compounds.Methods Based on the TCMSP and other databases,the composition library of Arnebiaeuchroma(Royle)Johnst.was constructed,the active molecules with better medicinal properties were screened by the admetSAR method,the targets were predicted and the network relationship was constructed based on the bSDTNBI method,and the key targets and pathways were analyzed by MCODE topology,the possible mechanisms of the main chemical components of Arnebiaeuchroma(Royle)Johnst and targets were verified by autoDOCK.A mouse model of blood-heat syndrome induced by 2,4-dinitrophenol(DNP,25 mg·kg^(-1))was established,measured the mouse body temperature,drinking water,urine volume,fecal water content and coagulation function indicators,and the behavioral system evaluated the sedative effect of representative compounds,including voluntary activities and body temperature,sleep effects,and open field experiment to analyze movement trajectory.Results According to the properties of ADME,OB 30%,DL>0.18 and CYP450 metabolic enzyme comprehensive scores were used to screen the top 20 active molecules,the main active representative molecule is shikonin(SK),mainly involved in neural ligand-receptor relationship and cancer pathways,etc.The main targets show that Arnebiaeuchroma(Royle)Johnst.is closely related to OPRK1,GABRG2 and other targets in the central nervous system.Further drug-target interaction points out that SK can be stably combined with GABRG2 and other targets.The results of in vivo pharmacodynamics showed that the amount of water in the SK group decreased,the amount of urine and the water content of feces increased,and the frequency of autonomous activities and body temperature of the mice were reduced significantly.In addition,SK significantly reduced the number of voluntary activities and body temperature in mice,and had dose-effect consistency,shortened the sleep latency,and significantly prolonged the sleep time induced by pentobarbital sodium(P<0.01);The experiment indicated that SK significantly reduced the total distance and average speed of DNP mice,and showed its less trajectory(P<0.01).Conclusion The medicinal molecules of Arnebiaeuchroma(Royle)Johnst.under the guidance of the heat-clearing effect are mainly naphthoquinone shikonin,which has a certain central antipyretic and partial central inhibitory effect.It is specifically manifested in that SK can effectively inhibit central excitement caused by blood heat.And this effect is related toγ-aminobutyric acid receptor postsynaptic inhibition and opioid receptor activation.