OBJECTIVETo investigate the cathartic effect of total anthraquinone (AQ) from rhubarb on SD rats and its regulatory effect on aquaporin 4 (AQP4) expression in rat colon and in vitro cultured LoVo cell line.

METHODSTwenty-four SD rats were randomly divided into the normal control group treated with distilled water, and the two AQ groups administered with AQ suspension in cathartic and high dose (AQcd and AQhd) respectively via gastrogavage for 5 days. Water content in colonic stool was detected and the expression of AQP4 in rat's proximal colon was measured using Western blot and RT-PCR. LoVo cells cultured in vitro were used in the experimental study. The AQP4 protein and mRNA expressions in the cells were detected by Western blot and semiquantitative RT-PCR after they were cultured for 24 h with RPMI-1640 medium containing rhein/emodin in different concentrations, and those cultured with RPMI-1640 containing 20 mg/L rhein/emodin for different time points.

RESULTSAfter treatment, the stool water content in the AQcd and AQhd groups was higher than that in the control group and the AQP4 expression in rats treated with AQ decreased in a dose-dependent manner. The study showed that rhein/emodin could significantly down-regulate the protein and mRNA expressions of AQP4 in cultured LoVo cells, with the effectiveness related with dose and acting time.

CONCLUSIONAt the same time of playing cathartic action, total AQ of rhubarb can effectively down-regulate the expression of AQP4 in rat's proximal colon; rhein/emodin can suppress the AQP4 expression in LoVo cells in vitro. One mechanism of cathartic effect of rhubarb AQ is possibly its down-regulation on AQP4 expression.

Animals ; Anthraquinones ; administration & dosage ; Aquaporin 4 ; genetics ; metabolism ; Cell Line, Tumor ; Colon ; drug effects ; metabolism ; Down-Regulation ; drug effects ; Gene Expression ; drug effects ; Humans ; Male ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rheum ; chemistry

OBJECTIVETo observe the effect of Tribulus terrestris extract on melanocyte stimulating hormone (MSH) expression in C57BL/6J mouse hair follicles, and investigate the role of Tribulus terrestris extract in activation, proliferation, epidermal migration of dormant hair follicle melanocytes.

METHODSThe aqueous extract of Tribulus terrestris was administered orally in specific pathogen-free C57BL/6J mouse at the daily dose equivalent to 1 g/1 kg in adult human, and the expression and distribution of MSH in the mouse hair follicles was observed with immunohistochemistry.

RESULTSThe positivity rate of MSH expression in the hair follicle melanocytes was 75% in mice treated with the extract, significantly higher than the rate of only 18.75% in the control group (P<0.01).

CONCLUSIONThe aqueous extract of Tribulus terrestris can significantly increase MSH expression in the hair follicle melanocytes by activating tyrosinase activity and promoting melanocyte proliferation, melanine synthesis, and epidermal migration of dormant melanocytes.

Administration, Oral ; Animals ; Cell Proliferation ; drug effects ; Female ; Hair Follicle ; cytology ; drug effects ; metabolism ; Immunohistochemistry ; Melanocyte-Stimulating Hormones ; biosynthesis ; Melanocytes ; cytology ; drug effects ; metabolism ; Mice ; Mice, Inbred C57BL ; Plant Extracts ; administration & dosage ; pharmacology ; Random Allocation ; Tribulus ; chemistry

OBJECTIVETo observe the effects of bacterial lysates (OM-85BV) and all trans-retinoic acid (ATRA) on airway inflammation in asthmatic mice, and to investigate the immunoregulatory mechanism of OM-85BV and ATRA for airway inflammation in asthmatic mice.

METHODSForty female BALB/c mice were randomly divided into five groups: normal control, model, OM-85BV, ATRA, and OM-85BV+ATRA. A bronchial asthma model was established by intraperitoneal injection of ovalbumin (OVA) for sensitization and aerosol challenge in all mice except those in the normal control group. On days 25-34, before aerosol challenge, the model, OM-85BV, ATRA, and OM-85BV+ATRA groups were given normal saline, OM-85BV, ATRA, and OM-85BV+ATRA respectively by gavage. Normal saline was used instead for sensitization, challenge, and pretreatment before challenge in the normal control group. These mice were anesthetized and dissected at 24-48 hours after the final challenge. Bronchoalveolar lavage fluid (BALF) was collected from the right lung to measure the levels of interleukin-10 (IL-10) and interleukin-17 (IL-17) by ELISA. The left lung was collected to observe histopathological changes by hematoxylin-eosin staining. The relative expression of ROR-γT mRNA was measured by quantitative real-time PCR.

RESULTSCompared with the normal control group, the model group showed contraction of the bronchial cavity, increased bronchial secretions, and a large number of infiltrating inflammatory cells around the bronchi and alveolar walls, as well as a significantly reduced level of IL-10 (P<0.05) and significantly increased levels of IL-17 and ROR-γT mRNA (P<0.05). Compared with the model group, the OM-85BV, ATRA, and OM-85BV+ATRA groups showed a significant reduction in infiltrating inflammatory cells around the bronchi and alveolar walls; the OM-85BV group showed a significant increase in the level of IL-10 in BALF (P<0.05) and significant reductions in the levels of IL-17 and ROR-γT mRNA (P<0.05); the ATRA group showed significant reductions in the levels of IL-17 and ROR-γT mRNA (P<0.05). Compared with the OM-85BV group, the OM-85BV+ATRA group had significantly increased relative expression of ROR-γT mRNA (P<0.05). Compared with the ATRA group, the OM-85BV+ATRA group had significantly increased levels of IL-10 and IL-17 in BALF (P<0.05).

CONCLUSIONSBoth OM-85BV and ATRA can reduce respiratory inflammation in asthmatic mice. However, a combination of the two drugs does not have a better effect than them used alone.

Animals ; Asthma ; drug therapy ; genetics ; immunology ; Cell Extracts ; administration & dosage ; Female ; Humans ; Interleukin-10 ; genetics ; immunology ; Interleukin-17 ; genetics ; immunology ; Lung ; drug effects ; immunology ; Mice ; Mice, Inbred BALB C ; Tretinoin ; administration & dosage

The present investigation was carried out to evaluate anti-inflammatory and membrane stabilizing properties of methyl jasmonate (MJ) in experimental rat models of acute and chronic inflammation. The effects of MJ on acute inflammation were assessed using carrageenan-induced rat's paw edema model. The granuloma air pouch model was employed to evaluate the effects of MJ on chronic inflammation produced by carrageenan in rats. The number of white blood cells (WBC) in pouch exudates was estimated using light microscopy. The levels of biomarkers of oxidative stress, such as malondialdehyde (MDA), glutathione (GSH) and activity of antioxidant enzymes in the exudates, were determined using spectrophotometry. The membrane stabilizing property of MJ was assessed based on inhibition of hemolysis of rat red blood cells (RBC) exposed to hypotonic medium. Our results indicated that MJ (25-100 mg·kg, i.p.) produced significant anti-inflammatory activity in carrageenan-induced paw edema in rats (P < 0.05). MJ reduced the volume of pouch exudates and the number of WBC in carrageenan-induced granulomatous inflammation. It also exhibited potent antioxidant and membrane stabilizing activities. In conclusion, these findings suggest the therapeutic potentials of methyl jasmonate in disease conditions associated with inflammation and its anti-inflammatory activity may be related to its antioxidant and membrane stabilizing activities.
Acetates ; administration & dosage ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; Cell Membrane ; chemistry ; drug effects ; immunology ; Cyclopentanes ; administration & dosage ; Disease Models, Animal ; Edema ; drug therapy ; immunology ; Erythrocytes ; chemistry ; drug effects ; Glutathione ; immunology ; Humans ; Male ; Malondialdehyde ; immunology ; Oxylipins ; administration & dosage ; Plant Extracts ; administration & dosage ; Rats ; Rats, Wistar

Animals ; Antineoplastic Agents, Alkylating/pharmacology* ; Breast Neoplasms/drug therapy* ; Cell Line ; Cell Line, Tumor ; Cyclophosphamide/pharmacology* ; Ginkgo biloba/chemistry* ; Mammary Neoplasms, Animal/drug therapy* ; Mice ; Plant Extracts/administration & dosage*

Cisplatin and other platinum-based drugs are used frequently for treatment of lung cancer. However, their clinical performance are usually limited by drug resistance or toxic effects. Carnosic acid, a polyphenolic diterpene isolated from Rosemary (Rosemarinus officinalis), has been reported to have several pharmacological and biological activities. In the present study, the combination effect of cisplatin plus carnosic acid on mouse LLC (Lewis lung cancer) xenografts and possible underlying mechanism of action were examined. LLC-bearing mice were treated with intraperitoneal injection with cisplatin, oral gavage with carnosic acid, or combination with cisplatin and carnosic acid, respectively. Combination of carnosic acid and cisplatin yielded significantly better anti-growth and pro-apoptotic effects on LLC xenografts than drugs alone. Mechanistic study showed that carnosic acid treatment boosted the function of CD8 T cells as evidenced by higher IFN-γ secretion and higher expression of FasL, perforin as well as granzyme B. In the meantime, the proportion of MDSC (myeloid-derived suppressor cells) in tumor tissues were reduced by carnosic acid treatment and the mRNA levels of iNOS2, Arg-1, and MMP9, which are the functional markers for MDSC, were reduced. In conclusion, our study proved that the functional suppression of MDSC by carnosic acid promoted the lethality of CD8 T cells, which contributed to the enhancement of anti-lung cancer effect of cisplatin.
Abietanes ; administration & dosage ; Animals ; Antineoplastic Agents ; administration & dosage ; CD8-Positive T-Lymphocytes ; drug effects ; immunology ; Carcinoma, Lewis Lung ; drug therapy ; genetics ; immunology ; Cell Line, Tumor ; Cisplatin ; administration & dosage ; Drug Synergism ; Humans ; Interferon-gamma ; genetics ; immunology ; Lung Neoplasms ; drug therapy ; genetics ; immunology ; Matrix Metalloproteinase 9 ; genetics ; Mice ; Mice, Inbred C57BL ; Myeloid-Derived Suppressor Cells ; drug effects ; immunology ; Plant Extracts ; administration & dosage ; Rosmarinus ; chemistry

AIMTo study the mechanism of evodiamine-induced cell death of A375-S2.

METHODSThe changes in cell morphology were observed by invert microscopy and Hoechst 33258 staining. DNA fragmentation was assayed by agarose gel electrophoresis. The effects of evodiamine on apoptosis and cell cycle were studied by flow cytometric analysis.

RESULTSEvodiamine was shown to markedly inhibit the growth of A375-S2 cells in dose- and time-dependent manners. At the early stage, evodiamine activated caspase cascades, which unexpectedly did not induce typical DNA fragmentation. At later stage, caspase inhibitors failed to block A375-S2 cell death induced by evodiamine. Evodiamine-induced cell death was shown to be not directly associated with cell cycle arrest.

CONCLUSIONAt the early stage, evodiamine initiates caspase-dependent and a typical apoptosis pathway in A375-S2 cells, but later it induces cell death through caspase-independent pathway which might be necrosis.

Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; Caspase Inhibitors ; Caspases ; metabolism ; Cell Cycle ; Cell Division ; drug effects ; DNA Fragmentation ; physiology ; Dose-Response Relationship, Drug ; Evodia ; chemistry ; Humans ; Melanoma ; enzymology ; pathology ; Plant Extracts ; administration & dosage ; isolation & purification ; pharmacology ; Quinazolines ; administration & dosage ; isolation & purification ; pharmacology ; Time Factors ; Tumor Cells, Cultured

OBJECTIVETo evaluate the therapeutic effect of MHSP65-TCL on melanoma and its effect on the activity of the immunocytes.

METHODSMHSP65-TCL was prepared by mixing MHSP65 with TCL derived from B16 melanoma cell lysate by repeated freezing and thawing. The MHSP65-TCL vaccine was administered in mice bearing B16 melanoma, and the changes in melanoma growth was observed. To investigate the influence of TCL in MHSP65-TCL on the activity of the immunocytes, we co-cultured TCL and mouse spleen cells in vitro, and analyzed CD69 expression on the cells, cell apoptosis, and levels of IL-10 and IFN-γ in the cell culture supernatant.

RESULTSThe MHSP65-TCL vaccine showed an anti-melanoma effect in the tumor-bearing mice. In the in vitro experiment, TCL in MHSP65-TCL strongly stimulated the activation of mouse spleen cells while causing apoptosis in some of the immunocytes and promoting cellular IL-10 secretion, but not IFN-γ.

CONCLUSIONSMHSP65-TCL derived from B16 melanoma cells has an anti-melanoma effect mediated by the activation of immunocytes. TCL in MHSP65-TCL also has immunosuppressive effect on immunocytes possibly due to the presence of suppressive components in TCL, and identifying and eliminating these components may potentially improve the anti-tumor actovoty of MSHP65-TCL vaccine.

Animals ; Antigens, CD ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; metabolism ; Apoptosis ; Bacterial Proteins ; administration & dosage ; immunology ; Cancer Vaccines ; Cell Extracts ; administration & dosage ; immunology ; Cell Line, Tumor ; Chaperonin 60 ; administration & dosage ; immunology ; Female ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Lectins, C-Type ; metabolism ; Melanoma, Experimental ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Random Allocation ; Spleen ; cytology ; immunology ; metabolism ; Tumor Burden ; immunology

AIMTo evaluate the effect of an ethanolic extract of Semecarpus anacardium fruits on spermatogenesis in albino rats.

METHODSMale albino rats were fed with a 50 % ethanolic extract of Semecarpus anacardium fruit at 100 mg.kg(-1).day(-1), 200 mg.kg(-1).day(-1) and 300 mg.kg(-1).day(-1) for 60 days. Fertility test was performed after 60 days of treatment. Sperm motility and density were observed in the cauda epididymis. Biochemical and histological analyses of the blood and reproductive organs were done. Recovery of fertility was followed to evaluate the reversibility of drug action.

RESULTSS. anacardium fruit extract administration resulted in spermatogenic arrest in albino rats. The sperm motility and density was reduced significantly. The RBC and WBC counts, haemoglobin, haematocrit, blood sugar and urea were found to be within the normal range in the whole blood. The protein, cholesterol and glycogen in the testes and the fructose in the seminal vesicle were significantly decreased after the treatment. The fruit extract feeding caused marked reduction in the number of primary spermatocytes, secondary spermatocytes and spermatids. The number of mature Leydig cells was also decreased and degenerating cells increased proportionately.

CONCLUSIONS. anacardium fruit extract causes spermatogenic arrest in albino rats.

Animals ; Cell Count ; Dose-Response Relationship, Drug ; Leydig Cells ; cytology ; Male ; Plant Extracts ; administration & dosage ; pharmacology ; Rats ; Semecarpus ; chemistry ; Sperm Count ; Sperm Motility ; drug effects ; Spermatogenesis ; drug effects ; Testis ; cytology ; drug effects ; metabolism

OBJECTIVETo investigate the effects of Coptis chinensis and Evodia rutaecarpa water extract on precancerous lesion of colon induced by DMH and proliferation and apoptosis changes of colon mucosa crypts.

METHODPrecancerous lesion of colon was induced by DMH. The changes of proliferation and apoptosis of colon mucosa crypts were detected by morphological analysis. The numbers of aberrant crypt foci (ACF) were measured by feulgen staining.

RESULTC. chinensis and E. rutaecarpa water extract could significantly inhibit the formation of ACF in model animals. The proliferative crypts were increased obviously in middle and distal colon, and decreased by C. chinensis and E. rutaecarpa water extract. The apoptosis crypts were increased in distal colon but not middle colon. C. chinensis and E. rutaecarpa water extract could promote apoptosis of both middle and distal colon.

CONCLUSIONC. chinensis and E. rutaecarpa water extract could significantly inhibit the formation of ACF in model animals. These results indicated that C. chinensis and E. rutaecarpa water extract maybe have an inhibitory and clinically therapeutic effect on colon cancer, which were partly resulted from inhibiting proliferation and promoting apoptosis of crypts in middle and distal colon.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; chemically induced ; drug therapy ; pathology ; physiopathology ; Coptis ; chemistry ; Dimethylhydrazines ; adverse effects ; Disease Models, Animal ; Evodia ; chemistry ; Humans ; Male ; Plant Extracts ; administration & dosage ; Random Allocation ; Rats ; Rats, Wistar