MAGED4B Promotes Glioma Progression via Inactivation of the TNF-α-induced Apoptotic Pathway by Down-regulating TRIM27 Expression / 神经科学通报·英文版

MAGED4B belongs to the melanoma-associated antigen family; originally found in melanoma, it is expressed in various types of cancer, and is especially enriched in glioblastoma. However, the functional role and molecular mechanisms of MAGED4B in glioma are still unclear. In this study, we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue, and the level was positively correlated with glioma grade, tumor diameter, Ki-67 level, and patient age. The patients with higher levels had a worse prognosis than those with lower MAGED4B levels. In glioma cells, MAGED4B overexpression promoted proliferation, invasion, and migration, as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide. On the contrary, MAGED4B knockdown in glioma cells inhibited proliferation, invasion, and migration, as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide. MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain. The interaction between MAGED4B and tripartite motif-containing 27 (TRIM27) in glioma cells was detected by co-immunoprecipitation assay, which showed that MAGED4B was co-localized with TRIM27. In addition, MAGED4B overexpression down-regulated the TRIM27 protein level, and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine (MG132), an inhibitor of the proteasome. On the contrary, MAGED4B knockdown up-regulated the TRIM27 level. Furthermore, MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132. Accordingly, MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7 (USP7) involved in the tumor necrosis factor-alpha (TNF-α)-induced apoptotic pathway. These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1 (RIP1)-dependent TNF-α-induced apoptotic pathway, which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.

Allosteric inhibition reveals SHP2-mediated tumor immunosuppression in colon cancer by single-cell transcriptomics

Colorectal cancer (CRC), a malignant tumor worldwide consists of microsatellite instability (MSI) and stable (MSS) phenotypes. Although SHP2 is a hopeful target for cancer therapy, its relationship with innate immunosuppression remains elusive. To address that, single-cell RNA sequencing was performed to explore the role of SHP2 in all cell types of tumor microenvironment (TME) from murine MC38 xenografts. Intratumoral cells were found to be functionally heterogeneous and responded significantly to SHP099, a SHP2 allosteric inhibitor. The malignant evolution of tumor cells was remarkably arrested by SHP099. Mechanistically, STING-TBK1-IRF3-mediated type I interferon signaling was highly activated by SHP099 in infiltrated myeloid cells. Notably, CRC patients with MSS phenotype exhibited greater macrophage infiltration and more potent SHP2 phosphorylation in CD68+ macrophages than MSI-high phenotypes, suggesting the potential role of macrophagic SHP2 in TME. Collectively, our data reveals a mechanism of innate immunosuppression mediated by SHP2, suggesting that SHP2 is a promising target for colon cancer immunotherapy.

Effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion-induced neuronal apoptosis / 临床麻醉学杂志

Objective To investigate protective effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion(OGD/R)-induced neuronal apoptosis.Methods SH-SY5Y cells were differentiated to neurons with ATRA and followed by TPA.According to the results of preliminary experiment, OGD/R modle was constructed by oxygen-glucose deprivation(OGD) for 12 h and reperfusion(R) for another 12 h.During the start of the OGD, neurons were immediately divided into six groups: group D0(0 μmol/L dexmedetomidine), group D1(0.1 μmol/L dexmedetomidine), group D2 (1 μmol/L dexmedetomidine), group D3 (10 μmol/L dexmedetomidine), group D4(100 μmol/L dexmedetomidine), group D5 (1 000 μmol/L dexmedetomidine).After reperfusion 12 h, the cell viability was evaluated by the method of MTT.The cellular apoptosis was observed by flow cytometry method.The protective effects of different concentration dexmedetomidine on OGD/R-induced neuronal apoptosis were investigated.Then in chosen the exact group having protective effects, endoplasmic reticulum stress specific protein mesencephalic astrocyte-derived neurotrophic factor (MANF) and pro-apoptotic protein Caspase-3 and CHOP were detected by Westernblot method.Results Compared with group D0, there was no difference on the cell viability and cellular apoptosis induced by OGD/R in groups D1 and D2, but a significant decrease and increase in groups D4 and D5 (P<0.01 or P<0.05).And only group D3 had a neuroprotective effect, significantly increased the cell viability and inhibited the apoptosis (P<0.01).Further studys found that group D3 significantly up-regulated ER stress specific protein MANF (P<0.01) and inhibited up-regulation of Caspase-3 and CHOP (P<0.01).Conclusion These data suggest that 10 μmol/L dexmedetomidine had neuroprotective effect on OGD/R-induced neuronal apoptosis and significantly increased cell viability.Our results also indicate that up-regulation of ER stress specific protein MANF and inhibition of CHOP and Caspase-3 by MANF are involved in the neuroprotective effects of Dexmedetomidine.

Autophagy alleviates neuronal toxicityinduced by abnormally phosphorylated tau protein / 中国药理学通报

Aim To observe the effects of rapamycin (Rapa) and starvation-induced autophagy on the morphology of neuronal cells, tau protein aggregation and expression of phosphorylated tau protein, to explore the possible mechanism of cytoprotective effect of these two classical autophagy inducers on phosphorylated tau expressing cells.Methods N2a cells were transfected with GFP-tau plasmid, and equal amount of empty vector was used as control.Then cells were incubated with or without okadaic acid(OA) for 12 h, followed by treatment with autophagy inducers rapamycin(Rapa) and EBSS, autophagy inhibitor Bafilomycin A1(Baf A1) for 6 h.DAB was used to observe tau expression and cell morphology.Confocal microscopy was used to observe the intracellular tau aggregation.TUNEL assay and cleaved caspase-3 level were used to detect cell apoptosis.Immunoblot was used to detect the expression of phosphorylated tau and autophagy-related proteins.Results Our study showed that the N2a cells treated with OA exhibited small cell body, retracted processes and increased tau aggregation, compared with only tau-expressing cells.Rapa and EBSS treatment significantly improved cell morphology, decreased tau aggregation and reduced cell apoptosis.On the contrary, Baf A1 treatment induced aberrant cell shape and increased tau aggregation and cell apoptosis.In addition, Rapa significantly decreased the high molecular weight, phosphorylated tau whereas EBSS especially decreased the low molecular weight phosphorylated tau.Conclusions Rapa and EBSS is alleviate hyperphosphorylated tau-induced cytotoxicity through different mechanism.Rapamycin mainly decreases phosphorylated tau oligomers, while EBSS liable to decrease the soluble phosphorylated tau.

Expression of RNF2 in breast carcinoma and its significance / 临床与实验病理学杂志

Purpose To investigate the expression of RNF2 in breast disease tissues and cell lines,and to analyze the association between expression of RNF2 and clinicopathological characteristics in breast carcinoma.Methods Expression of RNF2 protein and mRNA levels was detected using immunohistochemistry of EnVision two-step and qRT-PCR in breast carcinoma and benign breast disease as well as in cell lines.Results RNF2 expression was sigmificantly higher in breast carcinoma tissue specimens compared with benign breast disease specimens (P ＜0.05).Besides,the expression of RNF2 protein was significantly associated to tumor size,lymph node status and TNM stage (P ＜ 0.05 for both),but was not related to age,histological grade,the expression of ER,PR and HER-2 (P ＞ 0.05 for both).Higher expression of RNF2 mRNA was detected in breast carcinoma cell lines compared with breast epithelial cell lines (P ＜ 0.05).Conclusion RNF2 is overexpressed in breast carcinoma and can be a potential therapeutic target for breast carcinoma.

Effect of propofol on autophagy during oxygen-glucose deprivation/restoration in human liver cells / 中华麻醉学杂志

Objective To evaluate the effect of propofol on autophagy during oxygen-glucose deprivation and restoration (OGD/R) in human liver cells.Methods Human hepatic HL-7702 cells at the logarithmic growth phase were seeded into culture plates and randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),OGD/R group,and propofol + OGD/R group (group P+OGD/R).The cells were cultured in normal culture medium in group C.In OGD/R and P+OGD/R groups,the cells were subjected to O2-glucose deprivation for 6 h followed by restoration of O2-glucose supply for 12 h.Propofol with a final concentration of 50 mmol/L was added at 10 min before oxygen-glucose deprivation.The cell viability was detected by MTT assay.The expression of autophagy-related proteins such as microtubule-associated protein light chain 3 (LC3) and Beclin-1 was evaluated by Western blot.Immunofluorescence was used to determine the number and distribution of autophagosomes.Results Compared with group C,the cell viability was significantly decreased,the expression of LC3 and Beclin-1 was significantly up-regulated (P＜0.05),and the number of autophagosomes was significantly increased in OGD/R and P+OGD/R groups.Compared with group OGD/R,the cell viability was significantly increased,the expression of LC3 and Beclin-1 was significantly down-regulated (P＜0.05),and the number of autophagosomes was significantly decreased in group P+OGD/R.Conclusion The mechanism by which propofol reduces OGD/R injury is probably related to inhibition of autophagy in human liver cells.

Effects of phosphocreatine postconditioning on cerebral ischemia-reperfusion injury in rats / 临床麻醉学杂志

Objective To investigate the effects of phosphocreatine postconditioning on cerebral ischemia-reperfusion(IR)injury in rats.Methods Thirty-six SD rats were randomly divided into three groups:groups Sham,IR (treated with normal saline)and PCr.IR was induced by intraluminal middle cerebral artery occlusion (MCAO).All treatments were given intravenously at the begining of reperfusion.Twenty-four hours after the reperfusion, neurological deficit score and magnetic resonance scan were performed.serum concentrations of malonaldehyde and 4-hydroxynonenal,cere-bral infarct volume and destruction of cerebral cortex were estimated.Neuronal apoptosis was further assessed by immunohistochemistry and immunofluorescent staining of caspase-3 and NeuN. Results Compared with group IR,phosphocreatine significantly decreased neurological deficit score, infarct volume,malonaldehyde and 4-hydroxynonenal levels(P < 0.05 ).Cortex structure was more complete,as well as neuronal apoptotic index was smaller in group PCr (P <0.05).Conclusion PCr can reduce cerebral infarct volume,thereby promote neurofunctional recovery.The mechanism of Pcr is related to reduced oxidative stress and inhibitted apopotosis during IR.

Protective effects of MANF on oxygen-glucose deprivation/reperfusion-induced apoptotic ER stress in SH-SY5 Y neural cells / 中国药理学通报

Aim To investigate the protective effects of MANF on human neuroblastoma SH-SY5 Y cells suf-fering from oxygen-glucose deprivation/reperfusion ( OGD/R) and the underlying mechanism. Methods SH-SY5Y cells were treated with OGD for 6 h, fol-lowed by reperfusion for 12 h. Meanwhile, the cells were incubated with 2 μmol · L-1 recombinant human protein MANF for 12 h during reperfusion. The cell morphology was observed under an optical microscope. The cell viability was determined by MTT assay. PI staining was performed to detect the number of dead cells. Western blot was performed to determine the protein levels of endogenous MANF, glucose-related protein 78 ( GRP78/BiP) , phosphorylated inositol re-quiring enzyme 1 ( p-IRE1 ) , phosphorylated eukaryot-ic translation initiator factor 2α ( p-eIF2α) , cleaved caspase-3, and C/EBP-homologous protein (CHOP). Results The cells exposed to OGD/R became smaller and round, and the neurites of the cells were shortened or disappeared . Recombinant human protein MANF improved the survival rate ( P and cleaved caspase-3 in SH-SY5 Y cells induced by OGD/R were significantly suppressed by MANF. Con-clusion OGD/R up-regulates the ER stress-associated proteins and causes apoptosis. MANF inhibits OGD/R-induced cell death, which may be related to attenua-ting ER stress-induced apoptosis.

Effect ofα1-antitrypsin Z variant overexpression on autophagy regulation / 中国药理学通报

Aim To investigate the effect of α1-anti-trypsin Z variant (ATZ)overexpression on cell autoph-agy.Methods HEK 293T cells were transfected with pcDNA3.1 zeo+/ATM or pcDNA3.1 zeo+/ATZ,e-qual amount of empty vector was used as control.Cells were treated with NH4Cl for 4 hours and processed for detecting ATZ,LC3 and p62 by immunoblot.Mean-while ,expression and intracellular localization of ATZ, LC3 in 293 T cells were observed with double labeled immunofluorescence.The mRNA levels of autophagy-related genes were measured by real-time PCR.Immu-nohistochemistry was used to observe the morphology of ATZ-positive cells.Results Compared with the control,higher LC3Ⅱ levels and LC3 puncta were observed in ATZ transfected cells.Meanwhile,the levelsof p62 were decreased in ATZ transfected cells,andreversed by NH4 Cl (25 mmol·L -1 )treatment.Overexpression of ATZ increased the mRNA levels of Atg5and Atg12,but had no obvious influence on Beclin1.ATZoverexpressing cells presented abnormal morphologies.The nuclei became reduced,condensed,and even disappeared in ATZpositive cells.Conclusion ATZ overexpression increases autophagy activity whichmay be related to increasing Atg5 and Atg12 levels.

Effects of macrophage metalloelastase on angiostatin generation and growth of tumor vessels in murine colon carcinoma / 中华消化杂志

Objective To determine the pathway of macrophage metalloelastase (MME)generate active angiostatin by decomposing plasminogen and its effect on inhibiting growth of tumor and microvessel density (MVD) in vivo in mouse models. Methods The recombined plasmid pEGFPC1-MME was constructed. Thirty mice were subcutaneously inoculated with CT-26 cells that were stably transfected with pEGFP-C1-MME (MME-transfected group), 30 with CT-26 cells transfected with empty vector pEGFP-C1 (vector-transfected group) and 30 with CT-26 cells (non-transfected group). Radioiodination and radioisotope tracer were used to explore the pathway of angiostatin generation in vivo. Results SDS-PAGE electrophoresis analysis revealed that, in the PAGE gel contained the protein with molecular weights of 35 000 and 38 000, radioactivity in MME-transfected group was significantly higher than vector-transfected and non-transfected groups (P = 0. 00).Western blotting analysis demonstrated two bands containing 35 000 and 38 000 fragments in three groups. Quantification of the protein signals by image analysis revealed that the levels of 35 000 and 38 000 fragments were obviously increased in MME-transfected group (9.32±1.52 and 5.61±2.24,respectively) than those in vector-transfected (2.47 ± 0.23 and 0. 67 ± 0. 12, respectively) and nontransfected (1.21±0. 69 and 0. 86 ± 0.44, respectively) groups (P= 0.00). The average value of MVD and fluorescent express of vascular endothelial growth factor (VEGF) were lower in MMEtransfected group when compared with those in vector-transfected and non-transfected groups (P =0.00). The average tumor size in MME-transfected group was small in comparison with vectortransfected and non-transfected groups (P= 0.00). Conclusions MME is demonstrated to be one of matrix metalloproteinase that closely related with angiostatin production and has inhibitory effect on tumor growth in tumor-bearing mice.

Mechanisms of lumiracoxib on antitumor in human lung cancer cells / 中国药理学通报

Aim To investigate the possible mechanism of antitumor in human lung cancer cell lines A549 and NCI-H460 induced by lumiracoxib.Methods The expression of COX-2 was detected by Western blot and the levels of PGE2 and cAMP was determined by radioimmunoassay (RIA).Results COX-2 protein was highly expressed in A549 and NCI-H460 cells.After treatment with 15~240 ?mol?L-1 LUM for 24 hrs,LUM significantly decreased the level of COX-2 in A549 cells,but not in NCI-H460 cells.Compared with the control,the PGE2 production was reduced and the level of cAMP was increased after the treatment with 15,30,60,120,240 ?mol?L-1 of LUM,respectively.Conclusion The effect of Lumiracoxib on antitumor is in COX-2-dependent or-independent manner. The antitumor effect of LUM may be related to inhibiting the COX-2 activities by decreasing its secretion,up-regulating the level of cAMP,and down-regulating the level of PGE2.

The inflammatory and immune mechanisms following cerebral ischemia / 中国药理学通报

Hypoxia-ischemia induces an inflammatory and immune response in the nervous system that may be important for development of brain injury. Recent data implicate that cytokines, chemokines, adhesion molecules，are involved in the recruitment of inflammatory-immune cells. A cytokines which promote emigration of leukocytes from the vascular lumen into the injured brain tissue are produced at the site of incipient cerebral infarction. The expression of mRNA for alpha- and beta-chemokines preceded the appearance of immune cells suggesting that these molecules may have a role in the inflammatory response to insults. The humoral and cell-medated immunity under the conditions of focal brain ischemia was discussed. These results together indicate that cell-cell interaction by adhesion molecules and cytokines is an important component in the pathogenesis of ischemic cerebral diseases, especially at the acute phase.Recently accumulated data show that depleting the amount of circulating leukocytes or administering anti-inflammatory chemicals such as cytokine blocking agents, anti-adhesion molecule antibodies, and immunosuppressants effectively minimize the size of ischemia induced cerebral infarction.

Improvement of the cholinergic function by melatonin in amnesic rats induced by amyloid ?-peptide 25～35 / 中国药理学通报

AIM To investigate whether melatonin improve the learning and memory dysfunction in the amnesic rats induced by amyloid ? peptide 25～35 (A? 25～35 ) via cholinergic nervous system or not.Methods The amnesic model in adult rats was induced by injection of A? 25～35 into hippocampus; Morris water maze was used to determine the effects of A? 25～35 and melatonin on the learning and memory. The activity of the choline acetyltransferase and acetylcholinesterase were determined by immunohistochemistry and spectrophotometry respectively. RESULTS Injection of A? 25～35 20 ?ginto the adult rats hippocampus induced learning and memory dysfunction, and a decrease in the number of ChAT immunoreactive neurons in hippocampus. Melatonin (0 1, 1, and 10 mg?kg -1 , ig?10 d) improved the A? 25～35 treated rats cognitive function and increased the number of ChAT immunoreactive neurons in hippocampus. CONCLUSION Improvement of the cholinergic dysfunction by melatonin in adult rats induced by amyloid ? peptide 25～35 may be via cholinergic nervous system.

Induction of learning and memory dysfunction by ?-amyloid peptide fragment 25～35 in rats / 中国药理学通报

AIM To determine whether microinjection of ? amyloid peptide fragment 25～35 (A?25～35 ) into rat hippocampus induces the learning and memory dysfunction.METHODS Microinjection of A? 25～35 into hippocampus induced the rat learning and memory dysfunction. Step down test, Shuttle box test, and Morris water maze test were used to evaluate rat learning and memory function;The sections were stained with haematoxylin eosin (HE),the amyloid deposits were detected using Congo Red and silver.RESULTS The latency of A? 25～35 treated rats shortened,the stimulating time prolonged,and the numbers of errors increased in the step down test;Shuttle box test showed that A? 25～35 caused the latency and numbers of active escape reduction and the numbers of passive avoidance increase; Also in morris water maze, A?25 35 induced an impairment in rat spatial resolution; In A? 25～35 treated group, a significant decrease in the numbers of neurons in cortex and hippocampus,a massive glial reaction and neurophilic phenomenon were detected by HE staining; the positive vascular amyloidosis by Congo red and fibrils by Ag staining were also observed.CONCLUSION Microinjection of A? 25～35 into rat hippocampus may induce the learning and memory dysfunction and pathological changes which were similar to that found in Alzheimers disease (AD) brain.

Study on immunomodulation and antitumor activity of paeonol in HepA tumor mice / 中国药理学通报

AIM To study immunomodulation and antitumor activity of paeonol. METHODS Paeonol was administered singlely to HepA bearing mice. 14 d later, the inhibition of tumor weight were observed. IL-2 and TNF-? content in serum, IL-2 secretion by splencytes, TNF-? production by PM? were measured by RIA. RESULTS Paeonol possess markedly inhibitory activity on the growth of HepA tumor in mice, and IL-2 and TNF-? were induced signficantly. CONCLUSION Paeonol has the effects of antitumor. This effect may be derived from inducing IL-2 and TNF-? production.

Effects of selenium-enriched yeast on DTH in immunosuppressive mice / 中国药理学通报

The effects of selenium-enriched yeast (Sey) on DTH in immunosuppressive mice have been investigated. It was found that Sey (30, 60, 90 mg?kg-l?d-1?6 d, ig) was able to antagonize lowered DTH by cyclophospha-mide (Cy); Meanwhile, the thymus and spleen weights were increased by Sey (30, or 60 mg?kg-1?d-1) in immunosuppressive mice. Sey also potentiated the lowered ConA and LPS-indu-ced proliferation of splenocytes in DTH mice at the doses of 30, 60, or 90 mg ?kg-1 and 60 or 90 mg?kg-1 respectively. These results suggested that Sey could enhance the functions of lymphocytes.

The effects of recombinant human endostatin on adjuvant arthritis in rats / 中国药理学通报

AIMTo investigate the effects of the recombina nt human endostatin on adjuvant arthritis (AA) in rats and its mechanisms. METHODSThe model of rat AA was induced by injection of intradermal CFA. ConA and LPS induce d splencytes proliferation was examined by MTT asssy and the activities of inter leukin-1 (IL-1) and IL-2 were measured by the method of thymocytes prolifera tion. Synoviocytes were dispersed with incubation of collagenase and trypsin, an d IL-1, TNF production of synoviocytes was estimated with radio-immunity assay . RESULTSThe secondary inflammation of AA rats appeared on the 1 0th day after injection of CFA. The therapeutic administration of endostatin (0 1, 0 5, 2 5 mg?kg -1 ?d -1 , sc, ?7 d) was given at that time(d 10). It was found that endostatin significantly inhibited the secondary paw swel ling. The increased ConA-induced splenic lymphocyte proliferation reaction and the activated IL-1 and IL-2 activity of AA rats was reversed by the treatment with endostatin. Meanwhile,IL-1 produced by PM? was increased. Endostatin inh ibited IL-1 production from PM? and IL-1 and reduced TNF level of Synoviocyte s in AA rats. CONCLUSIONThe recombinant human endostatin has the rapeutical effect on AA rats, its mechanisms is related to its immunoregulative function.

Relation between animal models of RA and clinic / 中国药理学通报

Animal models of rheumatoid arthritis(RA)especially adjuvant arthritis(AA)in rats and collagen induced arthritis(CIA)in mice have much character similar to human RA in pathological mechanism,clinical representation and so on. The models are more ideal in investigating RA and selecting drugs of anti inflammation and immunity. To discuss the pathological mechanism and the relation between animal models of RA and clinic will be important not only to research RA in pathology,physiology,nervous endocrine immunity and so on,but also to find out safety and effective drugs of treatment RA. The articl summarizes several animal models of RA in establishment,pathological and clinical changes,inflammatory immune regulation and so on.

Studies on the analgesia and mechanisms of melatonin / 中国药理学通报

AIM To study the analgesic effects of melatonin (MT) and related mechanisms. METHODS The threshold was measured by hot plate test.? endorphine (? EP) was determined by radioimmunoassay (RIA). RESULTS MT (40～160 mg?kg -1 , ip) had significant dose and time dependent analgesic effects in hot plate test. The analgesia was also induced by injection (icv) of MT (0 25 mg?kg -1 ). The day night rhythm of pain threshold was not observed in 7 d after pinealectomy, but it appeared in sham operated rats.? EP content of rat brain measured by RIA indicated that the level of ? EP in hypothalamus and pituitary decreased significantly 1 h after ip MT(100 mg?kg -1 ). The analgesic effect of MT was attenuated by pretreatment of reserpine (3 mg?kg -1 ip) or ? receptor blocker phentolamine (100 mg?kg -1 , sc). CaCl 2 (230 mg?kg -1 ) antagonized the analgesic effect of MT, and EGTA and verapamil significantly enhanced the analgesia of MT in the same situation. CONCLUSION MT has significant analgesic action which may be related to opiate system, adrenergic system, and Ca 2+ content.

Determination of paeonol in the bulk drug and injection by RP-HPLC / 中国药理学通报

AIM To determine the concentration of paeonol in the bulk drug and injection, a method of reversed-phase high performance liquid chromatography (RP-HPLC) was developed. METHODS Lichrospher C_ 18 column(4.6 mm?250 mm, 5 ?m)was used with methanol-acetonitrile-water(30∶40∶30)as mobile phase at a flow rate of 0.8 ml?min -1 . 20 ?l of sample was injected into the C_ 18 column where the temperature was 25℃. The detection wavelength was 274 nm. RESULTS The linear regression equation for paeonol was = 3 772.525 8 X- 0.747 4 (r = 1.000 0, n =5) under the linearity range from 0.02 mg?L -1 to 25.60 mg?L -1 . The average recovery was 99.87%. The relative standard deviations of the intra-day and inter-day were less than 4%. CONCLUSION The method is simple and rapid, which may be used for the determination of paeonol and its preparation for different purpose.