OBJECTIVETo study the effects and mechanisms of Zuoguiyin (ZGY) on the ovarian nitric oxide (NO) production in peri-menopausal rats.

METHODSThe peri-menopausal model rats were respectively administered with low (13.78 g/kg), middle (20.67 g/kg), and high (31.00 g/kg) dose ZGY, and nilestriol for 8 weeks. Normal saline was given by gastrogavage to rats in the model group and the young control group (as the control group). The ovarian NO content and the activity of total nitric oxide synthase (NOS) were detected using nitrate reductase method and chemical colorimetry respectively. The mRNA and protein expressions of inducible NOS (iNOS), endothelial NOS (eNOS), and neuronal NOS (nNOS) were detected using RT-PCR and immunohistochemical assay.

RESULTS(1) Compared with that in the control group, the ovarian NO content and the activity of total NOS in peri-menopausal rats were significantly lower (P < 0.01). Middle and high dose ZGY could obviously up-regulate them (P < 0.01, P < 0.05). (2) The three kinds of NOS expression levels in perimenopausal rats were obviously lower when compared with those of the control group (P < 0.01). Middle dose ZGY could significantly promote all the three kinds of NOS expression levels of pre-senile rats (P < 0.01). High dose ZGY could up-regulate the expressions of iNOS and eNOS, while low dose ZGY could only enhance the iNOS expression (P < 0.01).

CONCLUSIONSThe down-regulated expressions of eNOS, iNOS, and nNOS in local ovaries resulted in decreased NOS activity and NO production, which were closely correlated with damaged microcirculatory vascular functions of ovaries in peri-menopausal rats. ZGY could protect rats' ovarian microcirculation by up-regulating the expressions of eNOS, iNOS, and nNOS, and enhancing the ovarian NOS activity and NO production.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Ovary ; drug effects ; metabolism ; Perimenopause ; Rats

OBJECTIVETo observe the effects of Xinfeng Capsule (XC) on the cardiac function and the myocardial ultrastructure in rats with adjuvant arthritis (AA).

METHODSSixty rats were randomly divided into the normal control group, the model group, the methotrexate (MTX) treated group, the Tripterygium Glycosides Tablet (TGT) treated group, and the XC treated group, 12 in each group. Except those in the normal control group, rats in the rest groups were induced to establish the AA model by intradermally injecting Freund's complete adjuvant into their right paws. The medication was started from the 19th day. Normal saline was administered to rats in the normal control group and the model group. MTX, TGT, and XC was respectively administered to rats in the MTX, TGT, and XC groups. The medication lasted for 30 days. The swelling degree of voix pedis, arthritis index (AI), the heart function, serum levels of cytokines, and the myocardial ultrastructure were observed.

RESULTS(1) Compared with the normal control group, the swelling degree of voix pedis and AI significantly increased (P < 0.01), the body weight significantly decreased (P < 0.05) in the model group and the other 3 treated groups. (2) Compared with the model group, the heart index (HI), left ventricular systolic pressure (LVSP), and left ventricular end diastolic pressure (LVEDP) significantly decreased (P < 0.05, P < 0.01), the maximum rate of left ventricular pressure of development or decline (+/- dp/dt(max)) significantly increased (P < 0.05, P < 0.01). Compared with the MTX treated group, the LVSP and LVEDP significantly decreased (P < 0.05), and +/- dp/dtmax significantly increased (P < 0.05). (3) Compared with the model group, TNF-alpha, brain natriuretic peptide (BNP), and IL-17 significantly decreased (P < 0.05, P < 0.01); IL-10, CD4+, CD4+ CDA25+ significantly increased (P < 0.05, P < 0.01). Compared with the MTX treated group, IL-17 significantly decreased (P < 0.05), CD4+ CD25+ expression significantly increased in the XC group (P < 0.05). (4) Transmission electron microscopy showed that myocardial ultrastructure was basically contact in the XC treated group, approaching to that of the normal control group.

CONCLUSIONSDecreased cardiac function and damaged myocardial ultrastructure existed in AA rats. XFC could ameliorate the swelling degree of voix pedis and AI, as well as improve the heart function. Its mechanisms might be correlated with down-regulating serum levels of inflammatory factors, up-regulating the expressions of anti-inflammation factors, thus improving the myocardial ultrastructure and protecting injured myocardial cells.

Animals ; Arthritis, Experimental ; drug therapy ; metabolism ; physiopathology ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Myocardium ; metabolism ; ultrastructure ; Phytotherapy ; Rats ; Rats, Wistar ; Tripterygium

OBJECTIVETo investigate the effects of intrathecal injection of ginsenoside Rg1 at different doses on the changes of the behavior and the expressions of excitatory amino-acid transporter 1 (EAAT1), i. e., glutamate-aspartate transporter (GLAST) in the spinal dorsal horn of the arthritis rats with chronic morphine tolerance, and further to explore its mechanisms for morphine tolerance.

METHODSAfter successful intrathecal injection, an adjuvant arthritis model was established in 36 healthy male SD rats. They were randomly divided into 6 groups, 6 in each group. They were intrathecally injected with 10 microL normal saline (Group NS), 10 microg morphine (Group M), 10 microg morphine + 50 microg ginsenoside Rg1 (Group MG50), 10 microg morphine +100 microg ginsenoside Rg1 (Group MG100), 10 microg morphine + 200 microg ginsenoside Rg1 (Group MG200), and 100 microg ginsenoside Rg1 (Group G100), respectively. The normal saline and morphine were intrathecally injected twice daily, while ginsenoside Rg1 at different doses was intrathecally injected once daily, for 7 successive days. Fifty percent mechanical paw withdrawal threshold (PWT) was dynamically detected to evaluate their behaviors. The rats were sacrificed on day 7 after medication. The L3-L5 segment of the spinal cord was isolated for determining the expression of GLAST in the spinal dorsal horn using immunofluorescence staining.

RESULTSThe PWT of Group M was significantly higher than that of Group NS on the 1st and 3rd day after medication (P < 0.05). But it was gradually shortened along with the increasing days of medication. There was no statistical difference between Group M and Group NS on the 7th day (P > 0.05), indicating the formation of morphine tolerance. The PWT of Group MG100 also showed a decreasing tendency, but obviously slower than that of Group M (P < 0.05). The PWT of Group G100 was higher than that of Group NS (P < 0.05). Compared with Group NS, the expression of GLAST in the spinal dorsal horn of rats in Group M was down-regulated (P < 0.01). Compared with Group M, the expression of GLAST in the spinal dorsal horn of rats in Group MG100 and Group G100 was up-regulated (P < 0.05).

CONCLUSIONSSingle application of ginsenoside Rg1 showed mild antinociceptive effect in adjuvant-induced arthritis rats. Intrathecal injection of 100 microg ginsenoside Rg1 could attenuate the formation of morphine tolerance. Its mechanisms might be correlated with up-regulating of the expression of GLAST.

Amino Acid Transport System X-AG ; metabolism ; Animals ; Arthritis, Experimental ; metabolism ; Drug Tolerance ; Ginsenosides ; administration & dosage ; pharmacology ; Injections, Spinal ; Male ; Morphine ; pharmacology ; Pain Measurement ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of ginsenoside Rb1 (GRb1) on the oxidative stress in the skeletal muscles of rats with postoperative fatigue syndrome (POFS) and to study its anti-fatigue mechanisms.

METHODSThe POFS model was established using resection of 70% of mid-small intestine. Ninety-six Sprague-Dawley (SD) rats were screened using grasping test. The rats were randomly divided into the control group, the model group, and the GRb1 treated group (at 10 mg/kg) by the body weight. The maximum grip strength of rats was detected on the 1st, 3rd, 7th, and 10th day after operation, respectively. The contents of malondialdehyde (MDA), the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-PX) were detected.

RESULTSCompared with the model group, the maximum grip strength was obviously enhanced on the postoperative day 7 and 10 (P < 0. 05), the MDA content obviously decreased on the postoperative day 3 and 7 (P < 0.05), the SOD activity obviously increased in the GRb1 treated group (P < 0.05). There was no obvious change in the activities of CAT and GSH-PX among the three groups at each time point (P > 0.05).

CONCLUSIONGRb1 could reduce the oxidative stress injury in the skeletal muscles, improve the activities of antioxidant enzymes, and enhance the functions of the skeletal muscles in POFS rats, which may be important reasons for fighting against POFS.

Animals ; Catalase ; metabolism ; Fatigue ; etiology ; metabolism ; Ginsenosides ; pharmacology ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; analysis ; Muscle, Skeletal ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Postoperative Period ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVETo explore the effects of Danzhi Jiangtang Capsule (DJC) and exercise on islet beta-cell function index (HOMA-% beta), blood glucose, and oxidative stress of diabetic rats.

METHODSA diabetic rat model was established using low dose streptozotocin and high fat forage in 60 male Wistar rats. The effects of exercise, DJC, and DJC combined exercise on the serum levels of fasting blood glucose (FBG), fasting insulin (FINS), triglyceride (TG), total cholesterol (TC), as well as the activities of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) in the pancreatic tissue were observed. The HOMA-%beta was also calculated. The main factors that affecting HOMA-%beta were explored using multi-factor regression analysis.

RESULTSCompared with model group, the levels of FBG, TG, TC, and pancreatic MDA were significantly reduced after intervention of exercise or DJC (P < 0.05, P < 0.01), while HOMA-% beta obviously increased (P < 0.01). The pancreatic GSH-Px activity significantly increased in the exercise group (P < 0.01). Exercise and DJC had synergistic effects on FBG, TG, HOMA-% beta, pancreatic SOD, and GSH-Px activities (P < 0.05). There was a negative and linear regression correlation between FBG and pancreatic SOD and GSHPx activities. HOMA-%beta was negatively correlated with FBG, TG, TC, and pancreatic MDA content, and positively correlated with SOD and GSH-Px activities. Besides, there was a linear regression correlation between HOMA-%beta and FBG.

CONCLUSIONExercise and DJC played synergistic effects, could improve the glucose and lipid metabolisms and enhance antioxidant activities, thus relieving the injury of pancreatic beta cells.

Animals ; Antioxidants ; metabolism ; Blood Glucose ; analysis ; Diabetes Mellitus, Experimental ; metabolism ; therapy ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Insulin-Secreting Cells ; drug effects ; metabolism ; Lipid Metabolism ; Male ; Oxidative Stress ; drug effects ; Pancreas ; drug effects ; metabolism ; Physical Conditioning, Animal ; Phytotherapy ; Rats ; Rats, Wistar

OBJECTIVETo observe the anti-tumor recurrent and metastatic efficacy of Ru'ai Shuhou Recipe (RSR) on HER2 positive breast cancer, to evaluate the effects of RSR on the expressions of matrix metalloproteinases (MMPs) and the tissue inhibitor of metalloproteinases (TIMPs) in the recurrence and metastasis of HER2 positive breast cancer, thus revealing its anti-tumor recurrent and metastatic mechanisms.

METHODSSelected were 30-week-old HER2/neu transgenic spontaneous breast cancer mice FVB/neu. The primary tumor resection was carried out. After surgery they were randomly divided into the blank control group, the RSR group, the Herceptin group, and the combination group (RSR + Herceptin group). The treatment lasted for 4 months. The inhibition rate of the recurrent tumor volume and the inhibition rate of the lung metastasis were evaluated. The expressions of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase (TIMP-1), and TIMP-2 in the recurrent tumor tissue were detected using Western blot.

RESULTSBy the end of the treatment the average recurrent tumor volume was 11.11 +/- 8.71 cm3 in the blank control group and 5.56 +/- 5.55 cm3 of the RSR group, showing statistical difference between the two groups (P = 0.037). The average lung metastatic nodule was 16 in the blank control group and 10 in the RSR group. The inhibition rate of lung metastasis was 37. 85% in the RSR group, but with no statistical significance. The expression level of activated MMP-2 in the RSR group was down-regulated when compared with the blank control group, the Herceptin group, and the combination group (P < 0.05). The expression of MMP-9 of the RSR group, the Herceptin group, and the combination group was significantly down-regulated when compared with the blank control group (P < 0.05). The expression of MMP-9 of the RSR group and the combination group was further down-regulated when compared with the Herceptin group (P < 0.05). The expressions of both TIMP-1 and TIMP-2 of the RSR group, the Herceptin group, and the combination group were all up-regulated when compared with the blank control group (P < 0.05). The increased expression of TIMP-1 was more significantly in the RSR group and the combination group when compared with the Herceptin group (P < 0.05). It was higher in the combination group than in the RSR group (P < 0.05).

CONCLUSIONSRSR could inhibit the tumor recurrence of FVB/neu mice. It could reduce the degradation of extracellular matrix and increase the protective effects of extracellular matrix. It might achieve its anti-tumor effect through effecting the invasive and metastatic capabilities of breast tumor cells.

Animals ; Breast Neoplasms ; drug therapy ; metabolism ; pathology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Phytotherapy ; Postoperative Period ; Receptor, ErbB-2 ; metabolism ; Tissue Inhibitor of Metalloproteinase-1 ; metabolism ; Tissue Inhibitor of Metalloproteinase-2 ; metabolism

OBJECTIVETo observe the cytotoxicity of indirubin derivative PHII-7 against human breast cancer MCF-7 cells and to study its primary mechanisms.

METHODSThe proliferation of MCF-7 cells was detected using MTT colorimetry. Annexin V/PI double staining was applied to detect the apoptosis rate of MCF-7 cells. The distribution of cell cycles was detected using PI staining and flow cytometry (FCM). The levels of reactive oxygen species (ROS) in MCF-7 cells were detected by DCFH-DA staining. The mRNA and protein levels of c-fos were detected using RT-PCR and Westem blot analysis.

RESULTSPHII-7 at different concentrations inhibited the proliferation of MCF-7 cells in a concentration-dependent manner, with the inhibitory rate ranging from 43.13% to 90.90% (P < 0.05). The inhibition was strengthened along with increased concentrations. PHII-7 at different concentrations could induce the apoptosis of MCF-7 cells. The early apoptosis rate was 1.43% +/- 0.02%, 9.14% +/- 0.36%, and 45.79% +/- 8.46%, respectively with the action of 1.25, 2.50, and 5.00 micromol/L PHII-7, respectively, showing dose-dependent manner. FCM analysis found that the proportion of MCF-7 cells in the G0/G1 phase and the S phase decreased after treatment with PHII-7, and the ratio of MCF-7 cells in the G2/M phase obviously increased (P < 0.01). The intra-cellular ROS level was significantly elevated 2 h after pretreatment with PHII-7. The levels of the protooncogene c-fos mRNA and protein were down-regulated in a dose-dependent manner after action of PHII-7.

CONCLUSIONSPHII-7 exerted obvious in vitro cytotoxic effects on MCF-7 cells. Its mechanisms might be associated with arresting the cell cycle, regulating the redox equilibrium, and down-regulating the expression of the protooncogene.

Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Female ; Humans ; Indoles ; pharmacology ; MCF-7 Cells

OBJECTIVETo investigate the mechanism of Shugan Jianpi Recipe (SJR) on the ion transportation of diarrhea predominant irritable bowel syndrome (IBS-D) colon mucosa induced by 5-HT.

METHODSTotally 36 male SD rats were randomly divided into three groups, i. e., the normal group, the model group, and the SJR group, 12 in each group. IBS-D Rat model was induced by intracolonic instillation of acetic acid and restraint stress. After successful modeling, normal saline was given to rats in the normal group and the model group, while SJR was given to those in the SJR group by gastrogavage for 14 days. The short circuit current (lsc) technology was used to measure 5-HT induced lsc changes of the colon mucosa under the actions of drugs and specific blocking agents.

RESULTSThere was no difference in basal current (BC), the potential difference (PD), and transmembrane resistance (TR) of the distal colon among the 3 groups (all P > 0.05). The 5-HT induced short circuit current change (delta lsc) was lower in the model group than in the normal group (P < 0.05), and it was higher in the SJR group than in the model group (P < 0.05). When 4,4'-diisothiocyanato-stilbene-2,2'-disulfonic acid (DIDS, 500 micromol/L), an Ca(+)-dependent Cl- channel blocker, was added from the epiphragm of the colonic mucosa, the 5-HT induced delta lsc was lower in the model group than in the normal group (P < 0.05), and it was higher in the SJR group than in the model group (P < 0.05). When Na+ was substituted in the epiphragm solution (Na+ free) or amiloride (100 micromol/L) was added from the epiphragm of the colonic mucosa, an epithelial Na+ channel blocker, the 5-HT induced delta lsc was lower in the model group than in the normal group (P < 0.05), and it was higher in the SJR group than in the model group (P < 0.05).

CONCLUSIONSSJR could affect the transmembrane electrolyte transportation of IBS-D rat induced by 5-HT through regulating the secretion of Cl- and HCO3-. The effects might be achieved by the coordination of apical Cl- channel CFTR, basolateral Na(+)-K+ ATPase, sodium-potassium-chloride cotransporter, sodium-bicarbonate cotransporter, Cl-/HCO3- exchanger, as well as K+ channel.

Animals ; Colon ; drug effects ; metabolism ; Diarrhea ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Intestinal Mucosa ; drug effects ; metabolism ; Ion Transport ; drug effects ; Irritable Bowel Syndrome ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the intervention of transcutaneous electrical acupoint stimulation (TEAS) on the renal blood flow at different levels of mean arterial pressure (MAP) in controlled hypotension.

METHODSForty-two male beagle dogs were randomly divided into seven groups, i. e., the general anesthesia group, the 50% controlled group, the 40% controlled group, the 30% controlled group, the 50% experimental group, the 40% experimental group, and the 30% experimental group, 6 in each group. Beagles in the general anesthesia group were not treated with controlled hypotension, and the target MAP was achieved in those of the rest groups and maintained for 60 min. In the experimental groups, TEAS was applied to bilateral Hegu (LI4), Zusanli (ST36), Sanyinjiao (SP6), and Quchi (LI11) at 2/100 Hz with the stimulation strength of (4 +/- 1) mA starting from the stability of their physiological conditions to 60 min of maintaining the target MAP level. The changes of the renal blood flow were monitored at different time points using laser Doppler.

RESULTSFrom starting pressure control to the target MAP level, the renal blood flow was significantly lower in the 30% controlled group than in the general anesthesia group and the basic level of the same group (P < 0.05), while there was no obvious change in the 30% experimental group. In maintaining the blood pressure, the renal blood flow was significantly lower in the 50% controlled group, the 40% controlled group, the 30% controlled group, and the 30% experimental group than in the general anesthesia group (P < 0.05), while there was no obvious change in the 50% experimental group or the 40% experimental group. By the end of blood pressure recovery, the renal blood flow restored to the basic level in the 50% controlled group, the 50% experimental group, and the 40% experimental group (P > 0.05), while it was not restored to the basic level in the 40% controlled group, the 30% controlled group, and the 30% experimental group (P < 0.05).

CONCLUSIONTEAS combined general anesthesia in controlled hypotension could effectively improve the renal blood flow, thus protecting the kidney.

Acupuncture Points ; Anesthesia, General ; methods ; Animals ; Dogs ; Hypotension, Controlled ; methods ; Male ; Renal Circulation ; Transcutaneous Electric Nerve Stimulation

OBJECTIVETo explore the effects of low-and high-frequency electroacupuncture (EA) on the gene expression profiles in rat spinal dorsal horn (DH) under the physiological state, thus providing the information to find out the differences of different EA frequencies induced effects.

METHODSUsing cDNA microarray, the changes of the gene expressions in the DH were detected and compared between 2 Hz EA and 100 Hz EA at bilateral Zusanli (ST36) and Sanyinjiao (SP6). The differentially expressed genes were identified. The EASE scores were used to comprehensively analyze the gene functions (by Gene Ontology) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways.

RESULTS(1) After EA stimulation 1 150 genes/expressed sequence tags (ESTs) were differentially expressed by 2 Hz EA, while 1 270 genes/ESTs were differentially expressed by 100 Hz EA. (2) Both 2 Hz and 100 Hz EA could induce the modulation of the same 516 genes/ESTs in the same direction, which was correlated with neural signal transmission. (3) The differentially expressed genes regulated specifically by 2 Hz were correlated with neural plasticity. (4) The differentially expressed genes regulated specifically by 100 Hz were correlated with stress and immunoregulation.

CONCLUSIONSEither low-or high-frequency EA could extensively regulate the spinal cord information processing. The low-frequency EA participated more in the regulation of neural plasticity, while high-frequency EA had more significant effects on stress and immunoregulation.

Animals ; Electroacupuncture ; Gene Expression ; Neuronal Plasticity ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Spinal Cord ; metabolism ; Transcription, Genetic ; Transcriptome