Bile duct ligation differently regulates protein expressions of organic cation transporters in intestine, liver and kidney of rats through activation of farnesoid X receptor by cholate and bilirubin

Body is equipped with organic cation transporters (OCTs). These OCTs mediate drug transport and are also involved in some disease process. We aimed to investigate whether liver failure alters intestinal, hepatic and renal Oct expressions using bile duct ligation (BDL) rats. Pharmacokinetic analysis demonstrates that BDL decreases plasma metformin exposure, associated with decreased intestinal absorption and increased urinary excretion. Western blot shows that BDL significantly downregulates intestinal Oct2 and hepatic Oct1 but upregulates renal and hepatic Oct2. In vitro cell experiments show that chenodeoxycholic acid (CDCA), bilirubin and farnesoid X receptor (FXR) agonist GW4064 increase OCT2/Oct2 but decrease OCT1/Oct1, which are remarkably attenuated by glycine-β-muricholic acid and silencing FXR. Significantly lowered intestinal CDCA and increased plasma bilirubin levels contribute to different Octs regulation by BDL, which are confirmed using CDCA-treated and bilirubin-treated rats. A disease-based physiologically based pharmacokinetic model characterizing intestinal, hepatic and renal Octs was successfully developed to predict metformin pharmacokinetics in rats. In conclusion, BDL remarkably downregulates expressions of intestinal Oct2 and hepatic Oct1 protein while upregulates expressions of renal and hepatic Oct2 protein in rats, finally, decreasing plasma exposure and impairing hypoglycemic effects of metformin. BDL differently regulates Oct expressions via Fxr activation by CDCA and bilirubin.

Mechanism of Tanreqing Capsule on treatment of coronavirus disease 2019 based on network pharmacology / 中草药

Objective: To explore the network regulation mechanism of Tanreqing Capsule (TRQC) on the treatment of coronavirus disease 2019 (COVID-19). Methods: Potential targets of the 19 major constituents in TRQC were predicted by the Swiss Target Prediction server and TCMSP database. Gene ontology (GO) function enrichment and pathway analysis of the targets were analyzed by Omicsbean analytic system and String 10 database. Finally, Cytoscape 3.6.1 software was used to construct the network pharmacology map. Results: A total of 19 compounds affected 68 pathways such as IL-17 signaling pathway, T cell receptor signaling pathway, arachidonic acid metabolism, cAMP signaling pathway, PI3K-Akt signaling pathway, influenza A, etc, by acting on 163 related targets, which associated with anti-inflammation, immune regulation, antipyresis, eliminating phlegm, relieving cough and asthma, analgesia, antibacterial and antiviral, and sedation. The network of "compound-target-pathway-pharmacological action-efficacy" was also constructed. Conclusion: The major constituents in TRQC, including scutellarin, baicalin, oroxylin-7-O-glucuronide, chrysin-7-O-glucuronide, forsythin, forsythiaside E, forsythiaside D, chlorogenic acid, caffeic acid, isochlorogenic acid A, ursodeoxycholic acid and chenodeoxycholic acid, may interfere with efficacy-related biological processes associated with antipyresis, eliminating phlegm and removing toxin by acting on key protein like TNF, EGFR, NOS3, PTGS2, IL2, GABBR1, MAPK14, ADRB2, REN, VCAM1, ACHE, PTPRC, etc.

Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS / 上海交通大学学报（医学版）

Objective : To develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma. Methods ¡¤ After extraction of C4, CA and CDCA with acetonitrile from plasma, they were quantified with stand-ard curve corrected by the internal standards based on Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Results ¡¤ The limits of detection (LOD) of C4, CA and CDCA were 0.16, 0.02 and 0.04 nmol/L respectively; All three metabolites had good linear relationships (correlation coefficients R2 were over than 0.998). The relative standard deviations (RSDs) of repeatabilities were below 3.0%. The RSDs of inter-day and intra-day precision were less than 6%, and the RSDs of stabilities at 4 °C were below 10% within 7 days. The average added recoveries of C4, CA and CDCA were 97.7%, 113.3% and 105.0%, respectively. Conclusion ¡¤ This method is of high detective sensi-tivity, good precision and stability, which meets the quantitative requirements of plasma biological samples.

Anti-inflammatory effect and mechanism of chenodeoxycholic acid on microglia cell BV2 induced by lipopolysaccharides / 中国药理学与毒理学杂志

OBJECTIVE: To explore the anti-inflammatory effect and mechanism of chenodeoxycholic acid (CDCA) on lipopolysaccharides (LPS)-induced microglia cell BV2. METHODS: BV2 cells were pre-treated with CDCA 25-100 μmol·L-1 for 2 h, and then incubated with LPS 200 mg·L-1 for 22 h. Nitric oxide (NO) content in the medium was detected by Griess reagent. Protein expressions of cyclooxygenase-2 (COX-2) and induced nitric oxide synthase (iNOS) were examined by Western blotting. Tumor necrosis factor-or (TNF-α), interleukin-6 (IL-6), IL-1β and G-protein coupled receptor 5 (TGR5) mRNA expressions were evaluated by quantitative PCR. BV2 cells were pre-treated with CDCA for 2 h, and then stimulated with LPS 200 mg·L-1 for 1 h. Phosphorylation of NF-ΚB, inhibitor of NF-ΚBα (IΚBα) and serine/threonine protein kinase (Akt) was analyzed by Western blotting, while nuclear translocation of NF-κB was examined by immunocytochemistry. RESULTS: Compared with the normal control group, LPS stimulation significantly increased the production of NO and the expressions of COX-2 and iNOS in cells of the model group (P<.05, P<0.01). Meanwhile, LPS stimulation elevated the mRNA expression levels of TNF-α, IL-6, and IL-1β (P<0.01) but decreased that of TGR5 (P<0.01). LPS stimulation increased the phosphorylation of NF-ΚB, IΚBα, and Akt (P<.05, P<0.01), which was accompanied by increased nuclear translocation of NF-KB. Compared with the model group, CDCA significantly reduced the NO content in the medium, and decreased the expression levels of COX-2 and iNOS (P< 0.01). It also significantly decreased the mRNA expression levels of TNF-α, IL-6, and IL-1β (P<0.01), but increased that of TGR5 (P<0.01). Further study disclosed that CDCA remarkably reduced the phosophorylation of NF-ΚB, IΚBα, and Akt (P<0.05, P<0.01) and nuclear translocation of NF-ΚB was observed. CONCLUSION: CDCA can significantly inhibit LPS-induced inflammation in BV2 cells, which might be related to the inhibition of Akt/NF-κB signaling pathway by activating TGR5.

Precise delivery of obeticholic acid

Primary bile acids were reported to augment secretion of chemokine (C‒X‒C motif) ligand 16 (CXCL16) from liver sinusoidal endothelial cells (LSECs) and trigger natural killer T (NKT) cell-based immunotherapy for liver cancer. However, abundant expression of receptors for primary bile acids across the gastrointestinal tract overwhelms the possibility of using agonists against these receptors for liver cancer control. Taking advantage of the intrinsic property of LSECs in capturing circulating nanoparticles in the circulation, we proposed a strategy using nanoemulsion-loaded obeticholic acid (OCA), a clinically approved selective farnesoid X receptor (FXR) agonist, for precisely manipulating LSECs for triggering NKT cell-mediated liver cancer immunotherapy. The OCA-nanoemulsion (OCA-NE) was prepared

Bile acids regulate anorexigenic neuropeptide through p-STAT3-SOCS3 signaling in mouse hypothalamic cells / 南方医科大学学报

OBJECTIVE@#To explore the effects of taurolithocholic acid (tLCA) and chenodeoxycholic acid (CDCA) on the expression of aorexigenic neuropeptide in mouse hypothalamus GT1-7 cells.@*METHODS@#Mouse hypothalamic GT1-7 cells were treated with culture medium containing 10% FBS (control group, =3) or with 10 nmol/L, 100 nmol/L, 1 μmol/L and 10 μmol/L tLCA (tLCA group, =3) or CDCA (CDCA group, =3) for 12, 24 or 48 h. Real-time PCR was performed to determine the expression levels of proopiomelanocortin (POMC) mRNA in the cells, and the production levels of α-melanocyte-stimulating hormone (α-MSH) were assessed using an ELISA kit. Signal transduction and activator of transcription 3 phosphorylation (p-STAT3), threonine kinase phosphorylation (p-AKT), suppressor of cytokine signaling 3 (SOCS3), G protein-coupled bile acid receptor-1 (TGR5) and farnesoid X receptor (FXR) protein were detected by Western blotting.@*RESULTS@#Western blotting results showed that mouse hypothalamic GT1-7 cells expressed two bile acid receptors, TGR5 and FXR, whose expressions were regulated by bile acids. Real-time PCR showed that the expression of POMC mRNA was significantly increased in the cells after treatment with 10 μmol/L tLCA or CDCA for 24 h. POMC-derived anorexigenic peptide α-MSH increased significantly in GT1-7 cells after treatment with 10 μmol/L tLCA or CDCA for 24 h. Treatment of the cells with tLCA or CDCA significantly increased the expressions of intracellular signaling proteins including p-STAT3, p-AKT and SOCS3.@*CONCLUSIONS@#Mouse hypothalamic GT1-7 cells express bile acid receptors TGR5 and FXR. Bile acids tLCA or CDCA can promote the expression of POMC mRNA and increase the production of the anorexigenic peptide α-MSH. The intracellular signaling proteins p-AKT, p-STAT3 and SOCS3 are likely involved in bile acid-induced anorexigenic peptide production.

Xantomatosis cerebrotendinosa sin xantomas tendíneos: caso clínico / Cerebrotendinous xanthomatosis: report of one case

Cerebrotendinous xanthomatosis (CTX) is an uncommon autosomal recessive disease caused by deficiency of 27-sterol-hydroxylase that results in an accumulation of cholestanol in the central nervous system, eyes, tendons, and blood vessels. We report a 22-year-old woman with a history of cataract surgery at the age of 14, cholecystectomy due to cholelithiasis at the age of 17 and chronic diarrhea, who presented with a six months period of gait instability and frequent falls. Physical examination revealed a bilateral pyramidal and cerebellar syndrome, with no visible tendon xanthomas. Cerebral magnetic resonance imaging showed an increase of the signal intensity on the T2-weighted images in periventricular cerebral white matter, dentate nuclei and spinal cord. With a high suspicion of CXT, a genetic study was conducted identifying a pathogenic variant in the CYP27A1 gene. There is considerable variation in clinical characteristics and age of onset of this disease, including absence of tendon xanthomas, delaying the diagnosis. Early recognition and chronic chenodeoxycholic acid therapy can improve outcome and quality of life.

The mechanism of chenodeoxycholic acid and its derivatives in hepatobiliary tumor / 临床肝胆病杂志

Chenodeoxycholic acid (CDCA) is a primary bile acid and is involved in the digestion,transportation,and absorption of nutriments in the hepatobiliary system.Farnesoid X receptor (FXR) is a member of the nuclear receptor superfamily.In vivo and in vitro studies have demonstrated that CDCA is the natural ligand for FXR and involved in many cell signaling pathways,and it can inhibit the proliferation and induce the apoptosis of hepatobiliary tumor cells.This article reviews the association between CDCA and hepatobiliary tumor.

Therapeutic strategies for patients with primary biliary cholangitis and suboptimal response to ursodeoxycholic acid / 临床肝胆病杂志

In recent years,ursodeoxycholic acid is commonly used for the treatment of primary biliary cholangitis (PBC);however,the growing number of PBC patients and the occurrence of suboptimal response and treatment intolerance pose a great challenge to treatment regimens.The approval of the new drug obeticholic acid brings hope to PBC patients,and a combination of fibrates also has a promising future.More studies are in progress.Although new drugs,such as monoclonal antibody,fibroblast growth factor 19,and sodium-dependent bile acid transporter inhibitor,have limited efficacy data,they provide new directions for the treatment of PBC.With the help of individualized follow-up and stratified therapy,the management of PBC patients will enter a new stage.

A case report on cerebrotendinous xanthomatosis.

Cerebrotendinous xanthomatosis (CTX) is a rare hereditary neuro-metabolic disease in which deposition of cholesterol and cholestanol occurs in various tissues including CNS. It is characterized by juvenile cataract, tendon xanthomas and progressive neurological defects. It is one of a group of neurologic disorder collectively referred to as leukodystrophy, which predominantly affects the CNS white matter. We are presenting a patient with cerebrotendinous xanthomatosis, who is now 36 years old, and shows the natural course of disease in an untreated patient. He presented with xanthomas on Achilles tendon, elbow and knees and showed cerebellar and pyramidal signs. He had recurrent seizures and was mentally subnormal.

Current developments in pharmacological therapeutics for chronic constipation

Chronic constipation is a common gastrointestinal disease severely affecting the patient׳s quality of life. The traditional treatment of constipation is the use of laxatives. Recently, several new drugs including lubiprostone, linaclotide and prucalopride have been approved for treatment of chronic constipation. However, a significant unmet medical need still remains, particularly among those patients achieving poor results by current therapies. The 5-HT4 receptor modulators velusetrag and naronapride, the guanylate cyclase C agonist plecanatide and the ileal bile acid transporter inhibitor elobixibat are recognized as the most promising drugs under investigation. Herein, we give a comprehensive review on the pharmacological therapeutics for the treatment of chronic constipation, with the purpose of reflecting the drug development trends in this field.

Inhibitory Effect of Chenodeoxycholic Acid-verticinone Ester on Tumor Growth of H22-bearing Mice / 医药导报

Objective To evaluate the antitumor effects of chenodeoxycholic acid-verticinone ester ( CDCA-Ver ) on tumor growth and immune system of H22-bearing mice. Methods Antitumor activity against a solid tumor mass was evaluated in Kunming mice. H22 cells were transferred into the abdomen cavity of Kunming mice. H22 cells were inoculated through subcutaneous injection at the right armpit of the mouse to establish a solid tumor model. At 24 h after H22 tumor cells inoculation, 40 tumor-bearing Kunming mice were randomly divided into 4 groups according to random number table ( n=10 each group):model control group, cyclophosphamide ( CTX) group, intraperitoneal CDCA-Ver injection group and intravenous CDCA-Ver injection group. In model control group, sterile 0. 9% sodium chloride solution (10 mL·kg-1 ) was intraperitoneally injected once daily. In CTX group and intraperitoneal CDCA-Ver injection group, CTX (20 mg·kg-1 ) and CDCA-Ver (20 mg·kg-1 ) was intraperitoneally injected once daily, respectively. In intravenous CDCA-Ver injection group, CDCA-Ver ( 20 mg · kg-1 ) was injected through tail vein once daily. CDCA-Ver, CTX and NS were injected into the mice of the experimental groups once daily for 10 days, respectively. The dose volume was 0. 1 mL · ( 10 g )-1 body weight. The positive control drug was cyclophosphamide. Ten mice were treated with 20 mg · kg-1 CDCA-Ver through intravenous injection ( i. v. ) . Ten mice were treated with 20 mg·kg-1 CDCA-Ver through intraperitoneal injection. The thymus and spleen indices and the tumor inhibition rate were assessed, and histopathological examination with haematoxylin and eosin ( H&E) staining was carried out to evaluate the antitumor effects of CDCA-Ver. Results CDCA-Ver ( ivor ip) suppressed the growth of solid tumor in H22-bearing mice. The inhibition rate was 48. 3% at the dose of 20 mg·kg-1 CDCA-Ver (ip). There was no significant difference between CDCA-Ver (ip) and CTX treated group (P<0. 05). Compared with the control, the weight of thymus and spleen of CDCA-Ver (ip) treated group was not obviously changed. But a significant weight loss of thymus and spleen in CTX group was observed, which was attributed to the immune suppression from CTX. The thymus and spleen indices in the CTX-treated mice were significantly lower than those of the control group (P<0. 01). We further conducted histopathological examination to confirm the results. The immune system was not suppressed by CDCA-Ver ( ip ) in tumor-bearing animals. The low toxicity of CDCA-Ver was an outstanding advantage for the development of newly anticancer drug. Conclusion CDCA-Ver treatment can significantly inhibit tumor growth in mice.

Bile acid nuclear receptor FXR and digestive system diseases

Bile acids (BAs) are not only digestive surfactants but also important cell signaling molecules, which stimulate several signaling pathways to regulate some important biological processes. The bile-acid-activated nuclear receptor, farnesoid X receptor (FXR), plays a pivotal role in regulating bile acid, lipid and glucose homeostasis as well as in regulating the inflammatory responses, barrier function and prevention of bacterial translocation in the intestinal tract. As expected, FXR is involved in the pathophysiology of a wide range of diseases of gastrointestinal tract, including inflammatory bowel disease, colorectal cancer and type 2 diabetes. In this review, we discuss current knowledge of the roles of FXR in physiology of the digestive system and the related diseases. Better understanding of the roles of FXR in digestive system will accelerate the development of FXR ligands/modulators for the treatment of digestive system diseases.

Role of farnesoid X receptor and bile acids in alcoholic liver disease

Alcoholic liver disease (ALD) is one of the major causes of liver morbidity and mortality worldwide. Chronic alcohol consumption leads to development of liver pathogenesis encompassing steatosis, inflammation, fibrosis, cirrhosis, and in extreme cases, hepatocellular carcinoma. Moreover, ALD may also associate with cholestasis. Emerging evidence now suggests that farnesoid X receptor (FXR) and bile acids also play important roles in ALD. In this review, we discuss the effects of alcohol consumption on FXR, bile acids and gut microbiome as well as their impacts on ALD. Moreover, we summarize the findings on FXR, FoxO3a (forkhead box-containing protein class O3a) and PPARα (peroxisome proliferator-activated receptor alpha) in regulation of autophagy-related gene transcription program and liver injury in response to alcohol exposure.

Efficacy of Magnesium Trihydrate of Ursodeoxycholic Acid and Chenodeoxycholic Acid for Gallstone Dissolution: A Prospective Multicenter Trial

BACKGROUND/AIMS: Cholecystectomy is necessary for the treatment of symptomatic or complicated gallbladder (GB) stones, but oral litholysis with bile acids is an attractive alternative therapeutic option for asymptomatic or mildly symptomatic patients. This study was conducted to evaluate the efficacy of magnesium trihydrate of ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) on gallstone dissolution and to investigate improvements in gallstone-related symptoms. METHODS: A prospective, multicenter, phase 4 clinical study to determine the efficacy of orally administered magnesium trihydrate of UDCA and CDCA was performed from January 2011 to June 2013. The inclusion criteria were GB stone diameter or =50%, radiolucency on plain X-ray, and asymptomatic/mildly symptomatic patients. The patients were prescribed one capsule of magnesium trihydrate of UDCA and CDCA at breakfast and two capsules at bedtime for 6 months. The dissolution rate, response rate, and change in symptom score were evaluated. RESULTS: A total of 237 subjects were enrolled, and 195 subjects completed the treatment. The dissolution rate was 45.1% and the response rate was 47.2% (92/195) after 6 months of administration of magnesium trihydrate of UDCA and CDCA. Only the stone diameter was significantly associated with the response rate. Both the symptom score and the number of patients with symptoms significantly decreased regardless of stone dissolution. Adverse events necessitating discontinuation of the drug, surgery, or endoscopic management occurred in 2.5% (6/237) of patients. CONCLUSIONS: Magnesium trihydrate of UDCA and CDCA is a well-tolerated bile acid that showed similar efficacy for gallstone dissolution and improvement of gallstone-related symptoms as that shown in previous studies.

Xantomatosis cerebrotendinosa: aspectos fisiopatológicos, clínicos y genéticos / Cerebrotendinous xanthomatosis: physiopathology, clinical manifestations and genetics

Cerebrotendinous xanthomatosis (CTX) is a rare autosomal recessive disease, caused by genetic deficiency of the 27-hydroxylase enzyme (encoded by CYP27A1). It plays a key role in cholesterol metabolism, especially in bile acid synthesis and in the 25-hydroxylation of vitamin D3 in the liver. Its deficiency causes reduced bile acid synthesis and tissue accumulation of cholestanol. Clinical manifestations are related to the presence of cholestanol deposits and include tendon xanthomas, premature cataracts, chronic diarrhea, progressive neurologic impairment and less frequently coronary heart disease, early onset osteoporosis and abnormalities in the optic disk and retina. An early diagnosis and treatment with quenodeoxycholic acid may prevent further complications, mainly neurological manifestations. This review summarizes cholesterol metabolism related to bile acid synthesis, physiopathology, biochemistry and treatment of cerebrotendinous xanthomatosis.

In vivo pharmacokinetics of major bioactive components from Tanreqing Injection in rats / 中草药

Objective: To investigate the in vivo pharmacokinetic characteristics of Tanreqing Injection in rats. Methods: Using baicalin (BC), ursodeoxycholic acid (UDCA), and chenodeoxycholic acid (CDCA) in Tanreqing Injection as biomarkers to characterize the in vivo pharmacokinetics. Rats were received an iv administration of Tanreqing Injection in single dose (1.25, 2.5, and 5.0 mL/kg) or multiple doses (5.0 mL/kg), and the blood concentration of BC was determined using LC-UV method, and the concentration of UDCA and CDCA was determined using LC-MS method. The pharmacokinetic parameters were analyzed by WinNonlin 6.3 software. Results: After a single iv administration of Tanreqing Injection (2.5 mL/kg) to rats, the values of t1/2 for BC, UDCA, and CDCA were (48.19 ± 12.74), (56.19 ± 33.33), and (109.96 ± 58.39) min, and AUC0-6h were (1718.02 ± 656.49), (1150.83 ± 371.53), and (541.52 ± 403.69) μg·min/mL. Within the tested dose range from 1.25 to 5.0 mL/kg, the values of t1/2 for the three major bioactive components were not affected by the dose, and AUC0-6h for BC and UDCA showed a good linearity to the dosage (r > 0.99), but AUC0-6h of CDCA was positively correlated with the dosage. After multiple iv administration of Tanreqing Injection (5.0 mL/kg), the values of AUC0-6h for BC, UDCA, and CDCA were (2872.37 ± 476.45), (3339.63 ± 939.94), and (1241.29 ± 408.38) μg·min/mL, and the accumulation index values were 83.3%, 118.5%, and 168.0%, respectively. Conclusion The elimination of BC and UDCA after single iv administration (1.25-5.0 mL/kg) is linear, and CDCA is probably linear after the administration. There is no significant accumulation in blood concentration for BC and UDCA, but CDCA has a tendency of accumulation after the multiple injection.

Effect of Tanreqing Injection in various formula compatibilities on pharmacokinetics of main components / 中草药

Objective: To study the pharmacokinetic effect of three active markers, baicalin (BC), ursodeoxycholic acid (UDCA), and chenodeoxycholic acid (CDCA) in plasma of rats after iv administration of Tanreqing Injection (TI) in various compatibilities, which contained scutellaria extraction (SE), bear gall powder extraction (BE), Cornu caprae hircus extraction (CE) and Lonicerae Flos extraction (FE), including SE, BE, SE-BE (SE and BE), SE-CE (SE and CE), SE-BE-CE (SE, BE and CE), and SE-BE-CE-FE (SE, BE, CE, and FE):. To discuss the rationality of TI compatibility, the effects of TI in various compatibilities on the pharmacokinetics of BC, UDCA, and CDCA in plasma of rats were investigated. Methods: Thirty-six experimental rats were randomly divided into six groups, and treated with SE, BE, SE-BE, SE-CE, SE-BE-CE, and SE-BE-CE-FE. After the simultaneous extraction of the three major bioactive components in plasma of rats, the concentration of BC was determined using LC-UV method, and UDCA as well as CDCA was determined using LC-MS method. The experimental data were analyzed by WinNonlin 6.3 software and the pharmacokinetic parameters of BC, UDCA, and CDCA in these recipes were evaluated. Results: The pharmacokinetic parameters of UDCA and CDCA did not change after iv administration of TI in various compatibilities. However, the TI in various compatibilities increased the AUC of BC after iv administration. The change degrees were SE < SE-CE < SE-BE-CE < SE-BE-CE-FE but with no statistical significance. The pharmacokinetic parameters of UDCA and CDCA did not obviously change, which indicated that the different compatibilities did not change the pharmacokinetic behavior of UDCA and CDCA. Conclusion: TI in various compatibelities could increase the exposure of BC following the iv administration, but could not affect the pharmacokinetic behaviors of UDCA and CDCA.

Apoptotic Effect of Co-Treatment with a Natural Product, Chios Gum Mastic, and a Synthetic Chenodeoxycholic Acid Derivative, HS-1200, on Human Osteosarcoma Cells / 체질인류학회지

Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. Recently it reported that CGM induced apoptosis in a few cancer cells in vitro. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM and a CDCA derivative, HS-1200 on human osteosarcoma (HOS) cells. To investigate whether the co-treatment of CGM and HS-1200 compared with each single treatment efficiently reduced the viability of HOS cells, MTT assay was conducted. Induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and DNA hypoploidy, Westen blot analysis and immunofluorescent staining were performed to study the alterations of the expression level and translocation of apoptosis-related proteins in co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, HOS cells co-treated with CGM and HS-1200 showed several lines of apoptotic manifestation whereas each single treated HOS cells did not. Although the single treatment of 40 microgram/mL CGM or 25 micrometer HS-1200 for 24 h did not induce apoptosis, the cotreatment of them induced prominently apoptosis. Therefore our data provide the possibility that combination therapy of CGM and HS-1200 could be considered as a novel therapeutic strategy for human osteosarcoma.