Danggui Buxue Tang (DBT) is a famous Chinese medicinal decoction. Mechanism of DBT action is wide ranging and unclear. Exploring new ways of treatment with DBT is useful. Sprague-Dawley(SD) rats were randomly divided into 3 groups including control (NC, Saline), the DBT (at a dose of 8.10 g), and blood deficiency(BD) (Cyclophosphamide (APH)-andCyclophosphamide(CTX)-induced anaemia). A metabolomics approach using Liquid Chromatography-Quadrupole-Time-of-Flight/Mass Spectrometry (LC/Q-TOFMS) was developed to perform the plasma metabolic profiling analysis and differential metaboliteswerescreened according to the multivariate statistical analysiscomparing the NC and BD groups, andthe hub metabolites were outliers with high scores of the centrality indices. Anaemia disease-related protein target and compound of DBT databases were constructed. The TCMSP, ChemMapper and STITCH databases were used to predict the protein targets of DBT. Using the Cytoscape 3.2.1 to establish a phytochemical component-target protein interaction network and establish a component, protein and hub metabolite protein-protein interaction (PPI) network and merging the three PPI networks basing on BisoGenet. The gene enrichment analysis was used to analyse the relationship between proteins based on the relevant genetic similarity by ClueGO. The results shown DBT effectively treated anaemia in vivo. 11 metabolic pathways are involved in the therapeutic effect of DBT in vivo; S-adenosyl-l-methionine, glycine, l-cysteine, arachidonic acid (AA) and phosphatidylcholine(PC) were screened as hub metabolites in APH-and CTX-induced anaemia. A total of 288 targets were identified as major candidates for anaemia progression. The gene-set enrichment analysis revealed that the targets are involved in iron ion binding, haemopoiesis, reactive oxygen species production, inflammation and apoptosis. The results also showed that these targets were associated with iron ion binding, haemopoiesis, ROS production, apoptosis, inflammation and related signalling pathways. DBT can promote iron ion binding and haemopoiesis activities, restrain inflammation, production of reactive oxygen, block apoptosis, and contribute significantly to the DBT treat anaemia.
Anemia ; blood ; chemically induced ; drug therapy ; metabolism ; Animals ; Chromatography, Liquid ; Cyclophosphamide ; toxicity ; Disease Models, Animal ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; therapeutic use ; Metabolic Networks and Pathways ; drug effects ; genetics ; Metabolome ; drug effects ; Metabolomics ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Tandem Mass Spectrometry

The aim of the present study was to explore the effect and mechanism of Bushen Huoxue recipe (BHR) on ovarian reserve in mice with premature ovarian failure (POF). Mice were divided into 3 groups: normal group, model group and BHR group. Intraperitoneal injection of cyclophosphamide was performed to create the POF model. Primordial follicular (PDF) number, ovarian wet weight, ovarian index, and estrous cycle were analyzed to evaluate the effect of BHR on POF. Meanwhile, the mRNA and protein level of Mouse Vasa Homologue (MVH) in the bone marrow, peripheral blood and ovary were detected, to explore the underlying mechanism of the treatment efficacy of BHR on ovarian reserve. By the time of BHR treatment for 28 days, BHR increased the PDF number and shortened the estrous cycle of POF mice. BHR also decreased the mRNA level of MVH in the bone marrow, and increased mRNA and protein level of MVH in the ovary of POF mice. Our results demonstrated a treatment efficacy of BHR on POF mice, and revealed that BHR might repair the dysfunction of germline stem cells in the bone marrow, and thus to improve the ovarian reserve and enhance the ovarian function of POF mice through neo-oogenesis.
Animals ; Bone Marrow ; drug effects ; metabolism ; Cyclophosphamide ; toxicity ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Estrous Cycle ; drug effects ; Female ; Gene Expression Regulation ; drug effects ; Humans ; Mice ; Ovarian Follicle ; drug effects ; growth & development ; Ovarian Reserve ; drug effects ; Primary Ovarian Insufficiency ; chemically induced ; drug therapy ; pathology

BACKGROUNDPhloroglucinol plays an important role in oxidative stress and inflammatory responses. The effects of phloroglucinol have been proven in various disease models. The aim of the present study was to investigate the efficacy and possible mechanisms of phloroglucinol in the treatment of interstitial cystitis (IC).

METHODSThirty-two female Sprague-Dawley (SD) rats were used in this study. IC was induced by intraperitoneal injection of cyclophosphamide (CYP). Rats were randomly allocated to one of four groups (n = 8 per group): A control group, which was injected with saline (75 mg/kg; i.p.) instead of CYP on days 1, 4, and 7; a chronic IC group, which was injected with CYP (75 mg/kg; i.p.) on days 1, 4, and 7; a high-dose (30 mg/kg) phloroglucinol-treated group; and a low-dose (15 mg/kg) phloroglucinol-treated group. On day 8, the rats in each group underwent cystometrography (CMG), and the bladders were examined for evidence of oxidative stress and inflammation. Statistical analysis was performed by analysis of variance (ANOVA) followed by least square difference multiple comparison post-hoc test.

RESULTSHistological evaluation showed that bladder inflammation in CYP-treated rats was suppressed by phloroglucinol. CMG revealed that the CYP treatment induced overactive bladder in rats that was reversed by phloroglucinol. Up-regulated tumor necrosis factor-α and interleukin-6 expression in the CYP-treated rats were also suppressed in the phloroglucinol treated rats. CYP treatment significantly increased myeloperoxidase activity as well as the decreased activities of catalase of the bladder, which was reversed by treatment with phloroglucinol.

CONCLUSIONSThe application of phloroglucinol suppressed oxidative stress, inflammation, and overactivity in the bladder. This may provide a new treatment strategy for IC.

Animals ; Cyclophosphamide ; toxicity ; Cystitis, Interstitial ; chemically induced ; drug therapy ; Female ; Inflammation ; drug therapy ; Oxidative Stress ; drug effects ; Phloroglucinol ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder ; drug effects ; pathology

OBJECTIVETo compare the effects and mechanism of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin and paeoniflorin.

METHODAlbiflorin and paeoniflorin were determined by using animal models of blood deficiency syndrome induced by cyclophosphamide. The amount of WBC, RBC, HGB, index of thymus gland and spleen, and the changes of GM-CSF, IL-3 and TNF-α in serum were detected after the treatment.

RESULTCompared with the model group, the amount of WBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01). The amount of RBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01, P < 0.001), which did not had a significant difference compared with the same dose. The index of thymus gland in the group of 30 mg x kg(-1) albiflorin was superior to the model group (P < 0.01), the difference was significant compared with the same dose of paeoniflorin (P < 0.05). The GM-CSF in serum in all groups of 30 mg x kg(-1) albiflorin, 15 mg x kg(-1) albiflorin, 30 mg x kg(-1) paeoniflorin and 15 mg x kg(-1) paeoniflorin increased obviously (P < 0.01, P < 0.05, P < 0.01, P < 0.05); The IL-3 in serum in both group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin also increased (P < 0.001). The content of TNF-α in group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were reduced (P < 0.01), which showed the obvious difference compared with the same dose group (P < 0.01).

CONCLUSIONAlbiflorin had the effect of blood enriching by regulating the immune function, same with the paeoniflorin. The probable mechanism of nourishing blood and liver of Paeoniae Radix Alba was not only the better effect of adjusting the content of TNF-α, but also might act synergistically with paeoniflorin.

Animals ; Blood Cells ; drug effects ; Bridged-Ring Compounds ; pharmacology ; Cyclophosphamide ; toxicity ; Glucosides ; pharmacology ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; Hematopoiesis ; drug effects ; Interleukin-3 ; blood ; Male ; Mice ; Monoterpenes ; pharmacology ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the effect of Morinda officinalis (MO) extract on cytoxan (CTX) -impaired spermatogenesis of adult male SD rats.

METHODSWe randomly divided 56 adult male SD rats into seven groups of equal number: blank control, CTX model, CTX + NS, CTX + 10 g/kg MO, CTX + 20 g/kg MO, CTX + 30 g/kg MO, and CTX + 40 g/kg MO. We made the models of impaired spermatogenesis in the SD rats by intraperitoneal injection of CTX and treated the animal models by intragastric administration of MO at the concentrations of 10, 20, 30, and 40 g per kg per d, respectively. After two weeks of medication, we observed the changes in the body weight, testicular and epididymal indexes, and microstructure of the testis tissue, measured the mean seminiferous tubule diameter (MSTD) , and obtained testicular biopsy scores (TBS) in different groups, followed by comparative analyses.

RESULTSAfter treatment, the CTX + NS group showed no remarkable differences in the body weight ([234.83 ± 28.77] g) and epididymal index (2.71 ± 0.34) from those of the four CTX + MO groups, but exhibited a significantly lower testicular index ([12.15 ± 1.04] g) than those in the CTX + 20 g/kg MO ([13.71 ± 0.97] g), CTX + 30 g/kg MO, ([13.30 ± 0.29] g), and CTX + 40 g/kg MO group ([13.48 ± 0.51] g) (P < 0.05). Light microscopy revealed obvious pathological changes of the testis tissue in the CTX + NS group and significantly ameliorated structures of the seminiferous tubules in the CTX + MO 10, 20, 30, and 40 g/kg groups, with the MSTD of (204.78 ± 11.03), (216.55 ± 10.93), (218.03 ± 11.23), and (218.59 ± 14.06) μm, respectively, and the TBS of 9.03 ± 0.39, 9.69 ± 0.26, 9.83 ± 0.18, and 9.89 ± 0.11, respectively, all significantly higher than (189.74 ± 8.55) μm and 5.95 ± 1.21 in the CTX + NS group (P < 0.05). The efficacy of MO extract was increased in a concentration-dependent manner.

CONCLUSIONMorinda officinalis extract can repair cytoxan-induced damage to rat spermatogenesis, with may achieve the best effect at the concentrations of 30 and 40 g per kg per d.

Animals ; Body Weight ; drug effects ; Cyclophosphamide ; toxicity ; Epididymis ; drug effects ; Male ; Morinda ; chemistry ; Mutagens ; toxicity ; Plant Extracts ; administration & dosage ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Seminiferous Tubules ; drug effects ; pathology ; Spermatogenesis ; drug effects ; Testis ; drug effects ; ultrastructure

OBJECTIVETo investigate the protective effect of epigallocatechin gallate (EGCG) on mouse sperm in vivo.

METHODSA total of 64 six-week-old male Kuming mice were randomly divided into eight groups of equal number to be treated with normal saline (negative control), Cyclophosphamide (CP) at 30 mg/kg (positive control), and CP followed by EGCG (experimental) at 20, 40, and 80 mg/kg, respectively, given every other day for 10 days. At 4 and 5 weeks after treatment, the bilateral testes of the mice were harvested for examination of sperm abnormality.

RESULTSEGCG did not increase the rate of CP-induced sperm abnormality in the mice, but reduced it instead with the prolonged time of treatment.

CONCLUSIONEGCG protects mouse sperm in vivo.

Animals ; Catechin ; analogs & derivatives ; pharmacology ; Cyclophosphamide ; toxicity ; Male ; Mice ; Mutagens ; toxicity ; Random Allocation ; Spermatozoa ; drug effects ; Time Factors

BACKGROUNDBone marrow hematopoietic function suppression is one of the most common side effects of chemotherapy. After chemotherapy, the bone marrow structure gets destroyed and the cells died, which might cause the hematopoietic function suppression. Heme oxygenase-1 (HO-1) is a key enzyme of antioxidative metabolism that associates with cell proliferation and resistance to apoptosis. The aim of this study was to restore or resist the bone marrow from the damage of chemotherapy by the HO-1 expression of mouse mesenchymal stem cells (mMSCs) homing to the mice which had the chemotherapy-induced bone marrow suppression.

METHODSOne hundred and sixty female Balb/c mice (6-8-weeks old) were randomly divided into four groups. Each group was performed in 40 mice. The control group was intraperitoneally injected for 5 days and tail intravenously injected on the 6th day with normal saline. The chemotherapy-induced bone marrow suppression was established by intraperitoneally injecting cyclophosphamide (CTX) into the mice which performed as the chemotherapy group. The mMSCs were tail intravenously injected into 40 chemotherapically damaged mice which served as the mMSCs group. The difference between the HO-1 group and the mMSCs group was the injected cells. The HO-1 group was tail intravenously injected into the mMSCs that highly expressed HO-1 which was stimulated by hemin. The expression of HO-1 was analyzed by Western blotting and RT-PCR. Cell proliferation was measured using the 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Histopathologic examinations were performed 1 week after injection.

RESULTSCompared with the control group, the expression levels of HO-1 mRNA and protein were significantly higher in the HO-1 group (all P < 0.05), even obviously than the mMSCs group. CTX treatment induced apoptosis and inhibited proliferation. After injected, the white blood cell (WBC), red blood cell (RBC) and platelet (PLT) declined fast and down to the bottom at the 7th day. The bone marrow structure was destroyed incomplete. In vitro, the survival rate of cells in chemotherapy group was less than 50% after 24 hours. In contrast, mMSCs could do a favor to the cellular cleavage and proliferation. They slowed down the cell mortality and more than 50% cells survived after 24 hours. The effects of blocking apoptosis and bone marrow recovery could be more effective in the HO-1 group. In the HO-1 group, it had observed that the bone marrow structure became complete and the hemogram closed to normal at 7th day.

CONCLUSIONSHO-1 played an important role in promoting the recovery of CTX-induced hematopoietic damage. We suggest that HO-1 is able to restore the functions of chemotherapy-induced hematopoietic damage.

Animals ; Apoptosis ; drug effects ; Blood Platelets ; drug effects ; Blotting, Western ; Bone Marrow ; drug effects ; enzymology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclophosphamide ; toxicity ; Erythrocytes ; drug effects ; Female ; Heme Oxygenase-1 ; genetics ; metabolism ; Leukocytes ; drug effects ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; enzymology ; physiology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo observe the effects of CMTM2 on cyclophosphamide (CP)-induced reproductive toxicity and the expression of steroidogenic acute regulatory (StAR) protein in the transgenic mouse model.

METHODSTwenty CMTM2 transgenic mice were equally divided into a CMTM2 + CP and a CMTM2 + NS group, the former intraperitoneally injected with CP at 50 mg per kg per d, while the latter with the equivalent dose of normal saline, both for 7 days. Another 20 wild C57BL/6J mice were randomly assigned to a WT + CP and a WT + NS group, treated the same way above. After 30 days, all the mice were sacrificed and their epididymides and testes removed for measurement of the serum testosterone level by radioimmunoassay, determination of sperm concentration and motility by light microscopy and detection of the expression of StAR by Western blot.

RESULTSThe levels of serum testosterone, sperm concentration and sperm motility were significantly decreased in the CMTM2 + CP group as compared with the CMTM2 + NS group ([42.98 +/- 3.25] nmol/L vs [46.74 +/- 3.38] nmol/L, [16.89 +/- 1.17 ] x 10(6)/ml vs [24.68 +/- 0.95 ] x 10(6)/ml, [72.75 +/- 1.25]% vs [85.14 +/- 1.12]%, P < 0.05), but remarkably less than in the WT + CP group ([37.97 +/- 4.17] nmol/L, [12.75 +/- 1.02] x 10(6)/ml, [50.52 +/- 1.37] %) (P < 0.05). However, the expression of StAR was significantly higher in the CMTM2 + CP than in the WT + CP group (1.16 +/- 0.07 vs 0.69 +/- 0.08, P < 0.05).

CONCLUSIONCMTM2 antagonizes cyclophosphamide-induced reproductive toxicity via regulating the expression of StAR, and hence plays a protective role in the reproductive system.

Animals ; Cyclophosphamide ; toxicity ; MARVEL Domain-Containing Proteins ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Repressor Proteins ; genetics ; Sperm Count ; Sperm Motility ; Testis ; drug effects ; metabolism

Interstitial lung disease in children (chILD) is a group of disorders characterized by lung inflammation and interstitial fibrosis. In the past recent years, we noted an outbreak of child in Korea, which is possibly associated with inhalation toxicity. Here, we report a series of cases involving toxic inhalational injury-associated chILD with bronchiolitis obliterans pattern in Korean children. This study included 16 pediatric patients confirmed by lung biopsy and chest computed tomography, between February 2006 and May 2011 at Asan Medical Center Children's Hospital. The most common presenting symptoms were cough and dyspnea. The median age at presentation was 26 months (range: 12-47 months), with high mortality (44%). Histopathological analysis showed bronchiolar destruction and centrilobular distribution of alveolar destruction by inflammatory and fibroproliferative process with subpleural sparing. Chest computed tomography showed ground-glass opacities and consolidation in the early phase and diffuse centrilobular nodular opacity in the late phase. Air leak with severe respiratory difficulty was associated with poor prognosis. Although respiratory chemicals such as humidifier disinfectants were strongly considered as a cause of this disease, further studies are needed to understand the etiology and pathophysiology of the disease to improve the prognosis and allow early diagnosis and treatment.
APACHE ; Bronchi/pathology ; Child, Preschool ; Cough/etiology ; Cyclophosphamide/therapeutic use ; Disinfectants/*toxicity ; Dyspnea/etiology ; Enzyme Inhibitors/therapeutic use ; Humans ; Hydroxychloroquine/therapeutic use ; Immunoglobulins/therapeutic use ; Infant ; Inhalation ; Lung Diseases, Interstitial/chemically induced/drug therapy/*pathology ; Prognosis ; Retrospective Studies ; Steroids/therapeutic use ; Tomography, X-Ray Computed

BACKGROUNDOveractive bladder (OAB) can be caused by many factors such as inflammation, bladder outlet obstruction, neurogenic factors. We performed an intraperitoneal (ip) injection of cyclophosphamide to induce cystitis in rats, which causes their detrusors to overact, to provide a valuable disease model for discussing OAB pathogenesis and to study effective curing methods.

METHODSFemale Sprague-Dawley rats were induced to form cystitis by cyclophosphamide (200 mg/kg, ip). The day after the injection, two catheters were inserted into each rat's bladder to study its urodynamics. The BL-410 model bio-function experimental system was used to monitor bladder pressure while the rats were conscious. Unstable detrusor contractions appear in the urine storage period as a standard to determine OAB, and the positive rate was calculated. Urodynamic parameters such as bladder basal pressure (BP), maximum voiding pressure (MVP), intercontraction interval (ICI), spontaneous activity (SA), maximum cystometric capacity (MCC), and bladder compliance (BC) were recorded in each group, and a light microscope was used to observe the pathological changes in the rat bladder tissue.

RESULTSThe detrusor instability rate of the model group was 83.33%. The MVP, MCC and BC of rats in the model group were lower than the control group (P < 0.01), and the BP, ICI and SA of the model group rats were higher than the control group (P < 0.01). The difference between the control group and the model group is statistically significant. The model group rats' bladder walls swelled and bled, the submucosa thickened and leukocyte infiltration became serious.

CONCLUSIONSAcute cystitis and OAB symptoms can be induced by ip injections of cyclophosphamide in rats. This can provide a valuable animal model to study OAB in human beings.

Animals ; Consciousness ; Cyclophosphamide ; toxicity ; Female ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder, Overactive ; chemically induced ; physiopathology ; Urodynamics ; physiology