In utero exposure to hexavalent chromium disrupts rat fetal testis development.

Hexavalent chromium (Cr6+) acts as an endocrine disruptor. Herein, we investigated effects of Cr6+ on the development of rat fetal Leydig and Sertoli cells, which support differentiation of the male reproductive tract in late gestation. Female pregnant Sprague Dawley rats were gavaged with potassium dichromate (0, 3, 6, and 12 mg/kg) from gestational days (GD) 12 to GD 21. Leydig and Sertoli cell function was evaluated by investigating serum testosterone levels, cell number and distribution, and the expression levels of Leydig and Sertoli cell genes and proteins. Cr6+ increased serum testosterone level at dose of 3 mg/kg (1.170 ± 0.121 ng/ml vs. 0.720 ± 0.082 ng/ml in the control), while lowered it at dose of 12 mg/kg (0.400 ± 0.098 ng/ml). In addition, it showed that Cr6+ dose-dependently reduced Leydig cell size and cytoplasmic size and decreased the percentage of medium fetal Leydig cell cluster at dose of 12 mg/kg. Further study demonstrated that the expression of Leydig cell (Lhcgr, Scarb1, and Hsd3b1) and Sertoli cell (Fshr, Pdgfa, and Lif) genes in the testis was upregulated at dose of 3 mg/kg while the expression of Lhcgr, Hsd17b3 and Igf1 was downregulated by Cr6+ at dose of 12 mg/kg. In conclusion, Cr6+ had biphasic effects on fetal Leydig cell development with low dose to stimulate testosterone production and high dose to inhibit it, possibly via biphasically regulating growth factor gene expression in fetal Sertoli cells.

Gestational exposure to ziram disrupts rat fetal Leydig cell development.

Ziram is an endocrine disruptor and may cause birth abnormality of the male reproductive system. However, the effects of ziram on fetal Leydig cell (FLC) development are still unknown. The objective of the present study was to determine the endocrine-disrupting effect of ziram on rat FLC development after gestational exposure. Pregnant Sprague Dawley dams were randomly divided into 5 groups and were gavaged with 0 (corn oil, the control), 1, 2, 4, or 8 mg/kg ziram from gestational day 12 (GD12) to GD21. FLC development was evaluated by measuring serum testosterone, FLC number and distribution, and the expression levels of Leydig and Sertoli cell genes. Ziram significantly increased serum testosterone level at 1 mg/kg (1.350 ±â€¯0.099 ng/ml vs. 0.989 ±â€¯0.106 ng/ml in the control), while it remarkably lowered it at 8 mg/kg (0.598 ±â€¯0.086 ng/ml). Quantitative immunohistochemical staining showed that ziram increased FLC number via stimulating cell proliferation at 1 mg/kg and lowered it via inhibiting its proliferation at 8 mg/kg without affecting Sertoli cell number. Further study demonstrated that the expression of Nr5a1, Lhcgr, Scarb1, Star, Cyp11a1, and Cyp17a1 genes and proteins in the testis was upregulated at 1 mg/kg and the expression of Leydig (Nr5a1, Lhcgr, Scarb1, Star, Cyp11a1, Cyp17a1, and Insl3) and Sertoli cell (Fshr, Hsd17b3, Dhh, Amh, and Sox9) genes and proteins was downregulated by ziram at 8 mg/kg. In conclusion, ziram had biphasic effects on FLC development with low dose to increase FLC number and function and high dose to decrease them.

Gossypol enantiomers potently inhibit human placental 3ß-hydroxysteroid dehydrogenase 1 and aromatase activities.

Gossypol is a chemical isolated from cotton seeds. It exists as (+) or (-) enantiomer and has been tested for anticancer, abortion-inducing, and male contraception. Progesterone formed from pregnenolone by 3ß-hydroxysteroid dehydrogenase 1 (HSD3B1) and estradiol from androgen by aromatase (CYP19A1) are critical for the maintenance of pregnancy or associated with some cancers. In this study we compared the potencies of (+)- and (-)-gossypol enantiomers in the inhibition of HSD3B1 and aromatase activities as well as progesterone and estradiol production in human placental JEG-3 cells. (+) Gossypol showed potent inhibition on human placental HSD3B1 with IC50 value of 2.3 µM, while (-) gossypol weakly inhibited it with IC50 over 100 µM. In contrast, (-) gossypol moderately inhibited CYP19A1 activity with IC50 of 23 µM, while (+) gossypol had no inhibition when the highest concentration (100 µM) was tested. (+) Gossypol enantiomer competitively inhibited HSD3B1 against substrate pregnenolone and showed mixed mode against NAD(+). (-) Gossypol competitively inhibited CYP19A1 against substrate testosterone. Gossypol enantiomers showed different potency related to their inhibition on human HSD3B1 and CYP19A1. Whether gossypol enantiomer is used alone or in combination relies on its application and beneficial effects.

[Curcumin improves the impaired working memory in cerebral ischemia-reperfusion rats by inhibiting proinflammatory cytokines].

OBJECTIVE: To investigate the ameliorative effect of curcumin pretreatment against impaired spatial working memory on global cerebral ischemia-reperfusion rats and to explore its mechanism. METHODS: After trained on a modified T-maze, 120 adult SD rats were randomly divided into 5 groups: sham group (S group), cerebral ischemia-reperfusion group (IR group), curcumin group (C group), LPS group (L group) and curcumin+LPS group (C+L group). Rats were treated with drugs or vehicles 1 h before 10 min global cerebral ischemia. Six rats in each group 7 days after reperfusion were tested in T-maze. Six rats in each group were sacrificed at 2 h, 1, 3 and 7 d after reperfusion and their serum or brains were harvested. Brain sections were stained with HE or toluidine blue and neuronal damage was quantified by the average neuronal density of CA1 area. Immunohistochemical staining for hippocampal IL-1ß and TNF-α was carried out, levels of serum IL-1ß and TNF-α was detected using ELISA procedure. RESULTS: Compared with S group, percentage of T-maze correct responses was decreased (88% ± 12% vs 69% ± 8%, P < 0.05), an extensive pyramidal neurons loss in CA1 area was observed, level of IL-1ß (0.26 ± 0.04 vs 0.53 ± 0.06, P < 0.05;48 ± 13 vs 161 ± 31, P < 0.05) and TNF-α (40.244 ± 0.025 vs 0.418 ± 0.036, P < 0.05; 33 ± 4 vs 85 ± 15, P < 0.05) in hippocampi or serum was increased in IR group. Compared with IR group, percentage of T-maze correct responses was increased (78% ± 13%) and average pyramidal neuronal density in CA1 area was increased with an decrease in hippocampi or serum IL-1ß (0.44 ± 0.09, 72 ± 19) and TNF-α (0.307 ± 0.047, 57 ± 14) in C group(P < 0.05). Compared with IR group, percentage of T-maze correct responses (61% ± 6%) was decreased with IL-1ß (0.86 ± 0.13, 331 ± 51), TNF-α (0.735 ± 0.059, 185 ± 20) in hippocampi and serum was increased in L group (P < 0.05). Compared with L group, percentage of T-maze correct responses (69% ± 12%) and average pyramidal neuronal density in CA1 area was increased with IL-1ß (0.69 ± 0.09, 246 ± 24), TNF-α(0.586 ± 0.047, 105 ± 25) in hippocampi and serum was decreased in C+L group (P < 0.05). CONCLUSION: Curcumin pretreatment improves the impaired spatial working memory in global cerebral ischemia-reperfusion rats by inhibiting proinflammatory cytokines.

Curcumin as a potent and selective inhibitor of 11ß-hydroxysteroid dehydrogenase 1: improving lipid profiles in high-fat-diet-treated rats.

BACKGROUND: 11ß-Hydroxysteroid dehydrogenase 1 (11ß-HSD1) activates glucocorticoid locally in liver and fat tissues to aggravate metabolic syndrome. 11ß-HSD1 selective inhibitor can be used to treat metabolic syndrome. Curcumin and its derivatives as selective inhibitors of 11ß-HSD1 have not been reported. METHODOLOGY: Curcumin and its 12 derivatives were tested for their potencies of inhibitory effects on human and rat 11ß-HSD1 with selectivity against 11ß-HSD2. 200 mg/kg curcumin was gavaged to adult male Sprague-Dawley rats with high-fat-diet-induced metabolic syndrome for 2 months. RESULTS AND CONCLUSIONS: Curcumin exhibited inhibitory potency against human and rat 11ß-HSD1 in intact cells with IC50 values of 2.29 and 5.79 µM, respectively, with selectivity against 11ß-HSD2 (IC50, 14.56 and 11.92 µM). Curcumin was a competitive inhibitor of human and rat 11ß-HSD1. Curcumin reduced serum glucose, cholesterol, triglyceride, low density lipoprotein levels in high-fat-diet-induced obese rats. Four curcumin derivatives had much higher potencies for Inhibition of 11ß-HSD1. One of them is (1E,4E)-1,5-bis(thiophen-2-yl) penta-1,4-dien-3-one (compound 6), which had IC50 values of 93 and 184 nM for human and rat 11ß-HSD1, respectively. Compound 6 did not inhibit human and rat kidney 11ß-HSD2 at 100 µM. In conclusion, curcumin is effective for the treatment of metabolic syndrome and four novel curcumin derivatives had high potencies for inhibition of human 11ß-HSD1 with selectivity against 11ß-HSD2.

[Curcumin down-regulates CX3CR1 expression in spinal cord dorsal horn and DRG in neuropathic pain rats].

OBJECTIVE: To investigate the effects of curcumin on the behavior of chronic constrictive injury (CCI) rats and the CX3CR1 expression in spinal cord dorsal horn and dorsal root ganglia (DRG). METHOD: Seventy-two male SD rats were randomly divided into 4 groups: 1) Sham operation group (Sham); 2) Chronic constrictive injury group (CCI); 3) Curcumin treated group (Cur), administrated with curcumin 100 mg x kg(-1) x d(-1) ip for 14 days after CCI; 4) Solvent contrast group (SC), administrated with an equal volume of solvent for 14 days after CCI. Paw thermal withdrawal (PTWL) and paw mechanical withdrawal threshold (PMWT) were measured on 2 pre-operative and 1, 3, 5, 7, 10, 14 post-operative days respectively. The lumbar segments L4-5 of the spinal cord and the L4, L5 DRG were removed at 3, 7, 14 days after surgery. The expression of CX3CR1 was determined by immunohistochemical staining. RESULT: Compared with Sham group, PTWL and PMWT in CCI group were significantly lower on each post-operative day (P<0.01), which reached a nadir on the 3rd day after CCI (PTWL was 6.5 +/- 1.1, PMWT was 22.6 +/- 5.1), and the expression of CX3CR1 were markedly increased in spinal cord dorsal horn and DRG. In Cur group, PTWL were higher than in CCI group on 7, 10, 14 post-operative day (P<0.05), and PMWT were higher than those in CCI group on 10 and 14 post-operative day (P<0.05). The administration of curcumin could significantly attenuate the activation of CX3CR1 induced by CCI. CONCLUSION: The study suggests that curcumin ameliorates the CCI-induced neuropathic pain, probably by attenuating the expression of CX3CR1 in spinal cord dorsal horn and dorsal root ganglia.

[Effect of curcumine on the nuclear pathway of JNK during hippocampal ischemia/reperfusion injury in SHR].

OBJECTIVE: To investigate the diversify of the nuclear pathway of c-Jun NH2-terminal kinases (JNK) during transient brain ischemia/reperfusion injury in hippocampal neuron apoptosis in spontaneously hypertensive rats (SHR) and to test whether the neuroprotection of curcumine on transient brain ischemia/reperfusion injury in SHR is related to the nuclear pathway of JNK. METHODS: Male Wistar-Kyoto (WKY) rats and SHR were randomly divided into five groups (n = 6): WKY sham group (W-Sham), WKY ischemia/reperfusion group (W-I/ R), SHR sham group (S-Sham), SHR ischemia/reperfusion group (S-I/R) and SHR curcumine (a chinese traditional medicine)100 mg/kg treatment group (S-Cur), which were sacrificed at 2 h, 6 h, 24 h, 3 d and 7 d after reperfusion. Global brain ischemic model was established by 4-VO method. The TdT-mediated dUTP nick end labeling (TUNEL) method was used to detect the neuron apoptosis in hippocampal CA1 region. The immunohistochemical method was applied to investigate the expressions of c-jun and c-fos in hippocampal CA1 region. RESULTS: The expressions of apoptosis and c-jun and c-fos in CA1 region in S-Sham group, W-I/R group and S-I/R group were more than those in W-Sham group (P < 0.05), were significantly increased in S-I/R group than those in W-I/R group (P < 0.05), and were significantly decreased in S-Cur group than those in S-I/R group (P < 0.05). CONCLUSION: Neuronal apoptosis and the expressions of c-jun and c-fos are more in SHR hippocampal. Global brain ischemia/reperfusion injury induces more expressions of apoptosis in hippocampal neuron in SHR, and the more expressions of c-jun and c-fos may participate in that process. The neuroprotection of curcumine in SHR is related to c-jun and c-fos.

[Effects of electroacupuncture on bispectral index and plasma beta-endorphin in patients undergoing colonoscopy].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on bispectral index (BIS) and plasma beta-endorphin (beta-EP) level in patients undergoing colonoscopy. METHODS: Sixty patients were equally randomized into EA group and control group with 30 cases in each. EA (2 Hz/100 Hz, 4-6 V) was applied to the right Zusanli (ST 36) and Shangjuxu (ST 37), and the left Yinlingquan (SP 9), Sanyinjiao (SP 6) and bilateral Hegu (LI 4) respectively 30 min before colonoscopy. The mean arterial pressure (MAP), heart rate (HR) and BIS in two groups were continuously monitored during the study. Plasma beta-EP concentration was detected by radioimmunoassay. The patient's adverse reactions (including pain, satisfaction degree, etc.) were evaluated by visual analog scale (VAS) and verbal stress scale (VSS). RESULTS: Self-comparison showed that MAP and HR in control group increased significantly during colonoscope's splenic flexure passing (P<0.05). Whereas the 2 indexes in EA group had no significant changes during colonoscope insertion, and its splenic flexure passing, hepatic flexure passing and post-enteroscopy (P>0.05). Comparison between two groups showed that MAP at the time-point of colonoscope insertion, and HR at the time-point of colonoscope's splenic flexure passing in EA group were significantly lower than those in control group (P<0.05). BIS values of EA group were significantly lower than those of control group at different time-points after colonoscope insertion (P<0.01). Plasma beta-EP concentrations at the time-points of colonoscope's hepatic flexure passing and post-enteroscopy were evidently increased in both groups in comparison with pre-enteroscopy (P<0.01), and beta-EP was significantly lower in EA group than that in control group at the time-point of colonoscope's hepatic flexure passing (P<0.05). The dosage of Midazolam used for conscious-sedation and the scores of VAS and VSS were also considerably lower in EA group than those in control group (P<0.05, P<0.01). No significant differences were found between two groups in the adverse reactions as dizziness, nausea, vomiting and abdominal pain, but the patients' satisfaction degree in EA group was evidently higher than that in control group (P<0.05). CONCLUSION: Acupuncture analgesia can effectively lower the colonoscopy patients' BIS value and plasma beta-EP level, meaning attenuation of the patients' stress responses during colonoscopy after EA.

Clinical pharmacology of cisatracurium during nitrous oxide-propofol anesthesia in children.

STUDY OBJECTIVE: To describe, in pediatric patients, the effects of three doses of cisatracurium during nitrous oxide-propofol anesthesia and to determine if larger doses result in faster onset time. SETTING: College hospital. SUBJECTS: 75 ASA physical status I and II children, aged 15 to 60 months, undergoing surgery for hypospadias or undescendent testis. INTERVENTIONS: Patients were randomly assigned to one of three groups according to the dose of cisatracurium: Group A = 0.1 mg/kg (two x effective dose), Group B = 0.15 mg/kg (three x effective dose), and Group C = 0.2 mg/kg (4 x effective dose). MEASUREMENTS: Neuromuscular block was assessed with TOF-Guard (Biometer International, Odense, Denmark) accelerometry. Onset time (from cisatracurium injection to maximal depression of time to first twitch), duration of peak effect (time from cisatracurium injection to 5% recovery of time to first twitch), duration of clinical action (time from cisatracurium injection to 25% recovery of time to first twitch), and recovery index (recovery of time to first twitch from 25% to 75%) were recorded. MAIN RESULTS: Cisatracurium had no effect on heart rate or blood pressure at any dose. Compared with Group A, onset times in Groups B and C were shorter; and durations of peak effect and clinical action in Groups B and C were longer (P < 0.01) than those in Group A. There was no difference in recovery index among the three groups. There was no difference in onset times between Groups B and C. Compared with Group B, durations of peak effect and clinical action in Group C were longer (P < 0.05 or P < 0.01). CONCLUSION: Four times the effective dose of cisatracurium did not significantly shorten onset time beyond that produced with three times the effective dose in young children.

[Application of acupuncture analgesia in colonoscopy].

OBJECTIVE: To evaluate the efficacy of acupuncture analgesia in reducing patient's discomfort during colonoscopy. METHODS: Eighty outpatients scheduled to undergo colonoscopy were randomly divided into an electroacupuncture (EA) group and a control group, 40 cases in each group. The patients in the EA group received electroacupuncture analgesia at the right Zusanli (ST 36) and Shangjuxu (ST 37), and left Yinlingquan (SP 9) and Sanyinjiao (SP 6), and bilateral Hegu (LI 4) from 30 minutes before colonoscopy to the end of colonoscopy. And the control group did not receive any treatment. Blood pressure (BP), heart rate (HR) in the both groups were continuously monitored during colonoscopy; the pain degrees during colonoscope inserting and passing the sigmoid, splenic flexure and hepatic flexure were observed. The time to reach cecum, adverse reaction and patient's satisfactoriness were recorded. RESULTS: Colonoscopy was well tolerated in all the 80 patients. Pain degrees during colonoscope inserting and passing the sigmoid and splenic flexure in the EA group were significantly lower than those in the control group (P < 0.01); the time to reach cecum in the EA group (9.58 +/- 3.386) min was significantly shorter than that in the control group (12.96 +/- 6.4) min (P < 0.05); patient's satisfactoriness in the EA group was significantly higher than that in the control group (P < 0.05). There was no significant difference in HR and BP values between the two groups. CONCLUSION: Acupuncture analgesia can effectively alleviate the discomfort of patients during colonoscopic examination, shorten the duration of colonoscopy, with a higher satisfactoriness of the patient.

Protective effect of curcumin on endotoxin-induced acute lung injury in rats.

To investigate the protective effect of curcumin on endotoxin-induced acute lung injury in rats, and explore the underlying mechanisms, 24 male Wistar rats were randomly divided into 4 experimental groups: sham-vehicle (S), sham-curcumin (C), lipopolysaccharide (LPS)-vehicle (L), and curcumin-lipopolysaccharide (C-L) groups. The wet/dry (W/D) weight ratio of the lung and bronchoalveolar lavage (BAL) fluid protein content were used as measures of lung injury. Neutrophil recruitment and activation were evaluated by BAL fluid cellularity and myeloperoxidase (MPO) activity in cell-free BAL and lung tissue. The levels of cytokine-induced neutrophil chemoattractant-I (CINC-1) in lung tissues were measured by ELISA. The histopathological changes of lung tissues were observed by using the HE staining. Our results showed that lung injury parameters, including the wet/dry weight ratio and protein content in BALF, were significantly higher in the L group than in the S group (P<0.01). In the L group, higher numbers of neutrophils and greater MPO activity in cell-free BAL and lung homogenates were observed when compared with the S group (P<0.01). There was a marked increase in CINC-1 levels in lung tissues in response to LPS challenge (P<0.01, L group vs S group). Curcumin pretreatment significantly attenuated LPS-induced changes in these indices. LPS caused extensive morphological lung damage, which was also lessened after curcumin pretreatment. All the above-mentioned parameters in the C group were not significantly different from those of the S group. It is concluded that curcumin pretreatment attenuates LPS-induced lung injury in rats. This beneficial effect of curcumin may involves, in part, inhibition of neutrophilic recruitment and activity, possibly through inhibition of lung CINC-1 expression.