Effects of Xiangsha Liujunzi decoction drug serum on gastric antrum smooth muscle cells from rats with functional dyspepsia by regulating gastrointestinal hormones

Objective:To observe the effect and mechanism of Xiangsha Liujunzi decoction(XSLJZD)drug serum on gastric antrum smooth muscle cells(SMCs)in rats with functional dyspepsia(FD).Methods:Gastric antrum SMCs from rats with FD were isolated,cultured,and then divided into six groups as follows:control,model,domperidone,low-dose XSLJZD(LXSLJZD),medium-dose XSLJZD(MXSLJZD),and high-dose XSLJZD(HXSLJZD).Each group was administered the corresponding drug serum for intervention.Drug serum intervention conditions and proliferative activity of SMCs were tested by cholecystokinin octapeptide.Ghrelin,gastrin,somatostatin,and substance P(SP)levels were measured by ELISA.Somatostatin and SP mRNA expression was measured by real-time PCR.Results:A concentration of 10%drug serum for 24 h was decided to be the best intervention condition for later study.The mean optical density value in the model group was lower than that in the control group(P紏.001).Optical density values in the domperidone and HXSLJZD groups were higher than those in the model group(P?.025,P?.032,respectively).Gastrin,SP,and ghrelin levels in the model group were lower(P?.007,P?.037,P?.005,respectively),but somatostatin levels were higher,compared with those in the control group(P?.031).Gastrin,SP,and ghrelin levels in the domperidone,MXSLJZD,and HXSLJZD groups were higher than those in the model group(all P<.05).Somatostatin levels in the four drug-treated groups were lower than those in the model group(P?.002,P?.007,P?.001,P?.009,respectively).SP mRNA levels in the model group were lower than those in the control,domperidone,MXSLJZD,and HXSLJZD groups(P?.037 P?.016,P?.025,P?.002,respectively).Somatostatin mRNA levels in the model group were higher than those in the control and MXSLJZD groups(P紏.042,P紏.035).Conclusions:XSLJZD and domperidone drug serum effectively promote proliferative activity of gastric antrum SMCs in an FD model.The mechanism of this activity may be regulated by gastrointestinal hormones.

Liujunzi decoction ameliorats cisplatin-induced anorexia via adjusting metabolic disorders in rats

OBJECTIVE Cisplatin is a formidable chemotherapy agent widely applying in antineoplastic treatments,but its side effects often limit the clinical usage.Metabolic disorders are one of the side effects induced by cisplatin,which closely relate to the onset of chemotherapy-induced anorexia(CIA)in cancer patients but lacks effective controls.Liujunzi decoction(LJZD)is a traditional Chinese formula that has a promising effect in treating CIA.However,whether LJZD ameliorates CIA through adjusting cisplatin-induced metabolic disorders remain unknow.The present study evaluated the mechanism of cisplatin-induced metabolic disorders,and the effect of LJZD in ameliorating these disturbances.METHODS 42 male Sprague-Dawley(SD)rats(180-220 g)were randomly divided into 3 groups:normal control group(distilled water+saline),model group(distilled water+cisplatin),LJZD group(4.8 g·kg^(-1)Liujunzi decoction ingredients+cisplatin).Intragastrical administered each drug twice a day(7∶00-19∶00)since day 0 for 4 d,animals were intraperitoneal injected with cisplatin 6 mg·kg^(-1)1 h after administration while normal control groups were injected with same volume of saline.On day 3,each group was anesthetized with pentobarbital sodium 45 mg·kg^(-1)(ip),and blood samples were collected from aorta abdominalis.Then the samples were analyzed using an LC-ESI-MS/MS system.Significantly regulated metabolites between groups were determined by VIP≥1 and absolute Log2FC(fold change)≥1.Identified metabolites were mapped to Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway database using Metaboanalyst 5.0(https://www.metaboanalyst.ca/).RESULTS A total of 133,77 and 32 differential metabolites were filtrated in control vs model,control vs LJZD and model vs LJZD groups respectively.Comparing to control,the levels of hexadecanoic acid(Log2FC=6.3153),linoleic acid(Log2FC=5.3478),and 8,11-icosadienoic acid(Log2FC=5.2342)significantly increased,and the levels of N-acetyl-L-tyrosine(Log2FC=-2.6283),cinnamic acid(Log2FC=-2.3381),N-acetylphenylalanine(Log2FC=-2.2501)significantly decreased in model group.The KEGG pathway enrichments of these metabolites indicated that,cisplatin-induced metabolic disorders by disturbing metabolism pathways such as linoleic acid metabolism,biosynthesis of unsaturated fatty acids,and phenylalanine metabolism,which suggested that the onset of CIA was partly associated with the metabolic disorders of linoleic acid,unsaturated fatty acids,and phenylalanine.Compared to control,treatment of LJZD significantly increased the levels of 4-hydroxytryptamine(Log2FC=12.0186),hexadecanoic acid(Log2FC=5.7412),linoleic acid(Log2FC=5.1877)and significantly decreased the levels of N-acetylmethionine(Log2FC=-1.7317),2-aminoethanesulfinic acid(Log2FC=-1.6578),N-acetyl-L-tyrosine(Log2FC=-1.5355).And comparing to the model group,4-hydroxytryptamine(Log2FC=12.0186),7,12-diketocholic acid(Log2FC=2.0998),N-acetylneuraminic acid(Log2FC=2.0560)markedly increased,and 3-hydroxy-3-methylpentane-1(Log2FC=-1.9202),5-dioic acid(Log2FC=-1.7166),N-isovaleroylglycine,hexanoyl glycine(Log2FC=-1.4958)markedly decreased in LJZD group.It was worth noting that,there were 23 differential metabolites filtrated both in control vs model and model vs LJZD groups,which were the key metabolites of LJZD in treating CIA.Among these 23 common metabolites,there were 16 metabolites excluding the control vs LJZD group,that was,LJZD had no effect in normal rats while being able to ameliorated cisplatin-induced metabolic disorders by regulating these 16 metabolites.Cisplatin-induced downregulation of 11 metabolites such as hydrocinnamic acid,(±)12(13)epoxy-9Z-octadecenoic acid,cinnamic acid were upregulated after LJZD treatment,and cisplatin-induced upregulation of imidazoleacetic acid,2′-deoxycytidine-5′-monophosphate and other 5 metabolites were downregulated by LJZD.The KEGG pathway analysis indicated that the linoleic acid metabolism,histidine metabolism,and pyrimidine metabolism were the most enriched metabolic pathway.Thus,cisplatin-induced metabolic disturbances mainly by disturbing linoleic acid metabolism,histidine metabolism,and pyrimidine metabolism,and LJZD interacted with these metabolic pathways to reduce metabolic disorders and thus ameliorated CIA.CONCLUSION Cisplatin-induced anorexia was closely related to the metabolic disorders of linoleic acid metabolism,biosynthesis of unsaturated fatty acids,and phenylalanine metabolism.The mechanism of LJZD in ameliorating CIA was in concerned with the metabolic adjustments,relating to the regulation of linoleic acid metabolism,histidine metabolism,and pyrimidine metabolism.

Mechanism underlying the effect of Liujunzi decoction(六君子汤)on advanced-stage non-small cell lung cancer in patients after first-line chemotherapy

OBJECTIVE:To further clarify the anticancer mechanisms of Liujunzi decoction(六君子汤)and provide possible targets for the treatment of advanced-stage nonsmall cell lung cancer(NSCLC)by re-analyzing differential gene expression profile of peripheral blood mononuclear cells(PBMCs)from Liujunzi decoctiontreated NSCLC patients receiving first-line chemotherapy.METHODS:The PBMC gene expression microarray data set GSE61926 was retrieved from a high throughput gene expression database.Differentially expressed genes(DEGs)were screened by paired sample t-test and the multiple ratio method.Gene ontology and Kyoto encyclopedia of genes and genomes(KEGG)pathway analyses were performed using the DAVID database.The protein–protein interaction(PPI)network was constructed using interaction gene library retrieval tools and Cytoscape software.RESULTS:A total of 162 DEGs were identified,with 67 upregulated genes and 95 downregulated genes.The functional distribution of Gene Oncology(GO)genes showed that DEGs were mostly concentrated in extracellular regions,calcium ion binding,and transcriptase activity.KEGG pathway analysis showed that cytokine–cytokine receptor interactions were significantly enriched.PPI network analysis screened out the top 10 central protein-coding genes with the highest nodal degree:IL2,PIWIL4,DICER1,PIWIL2,SAA1,XCL1,IL22RA1,ARHGAP11A,DCP1A,and GDNF.Among them,the central protein-coding gene with the highest node degree was IL2.In addition,the central protein-coding genes with high node degrees and high molecular complex detection(MCODE)scores were PIWIL4,DICER1,PIWIL2,and DCP1A,all of which are related to tumor development.CONCLUSIONS:One signaling pathway and 10 central protein-coding genes related to anticancer mechanisms were screened by re-analysis of GSE61926 data.IL2,PIWIL4,DICER1,PIWIL2,and DCP1 A may have important roles in the mechanism of Liujunzi decoction treatment against NSCLC.Our results suggest that the anticancer mechanism of Liujunzi decoction may be related to gene silencing by RNA and the biological processes of piwi-interacting RNA and other small RNAs.