ABSTRACT
The purpose of this study was to investigate how Zuogui pills from the Kidney-tonifying and Nourishing Yin formula, in combination with the gonadotrophin-releasing hormone antagonist cetrorelix, affected the ovarian local oxidative stress response in decreasing ovarian reserve (DOR) mice. All animal experiments were carried out in accordance with the guidelines and standards established by Jinan University's Experimental Animal Management Committee. Cyclophosphamide (CTX)-treated DOR mice were given Zuogui pills, cetrorelix, or a combination of the two drugs intragastrically. After treatment, there were changes in the estrous cycle, serum sex hormone levels, oxidative stress-related indexes, growth biochemical factor levels, and SIRT1/P53/P21 expression. In comparison to the model group, the Zuogui pills and the cetrorelix+Zuogui pills group had significantly prolonged estrous periods and shortened interestrous periods, and the cetrorelix+Zuogui pills group had a significantly shortened cycle length. Follicle-stimulating hormone (FSH) decreased and estradiol (E2) increased in all treatment groups compared to the model group, oxidative stress indexes nitric oxide synthase (NOS), nitric oxide (NO), and reactive oxygen species (ROS) decreased, growth biochemical factors brain derived neurotrophic factor (BDNF) and growth differentiation factor 9 (GDF-9) concentrations increased significantly, and leukemia inhibitory factor (LIF) showed no significant change. SIRT1/P53/P21 immunohistochemical results revealed that, when compared to the model group, the expression of SIRT1 increased while the expression of P53 and P21 proteins decreased in all treatment groups, with the cetrorelix+Zuogui pills group having the largest decrease, with significant differences in all indicators. We conclude that cetrorelix combined with Zuogui pills for kidney nourishing and Yin recipe improved the oxidative stress response in the follicle by regulating the SIRT1/P53/P21 pathway, reducing peroxide product production, protecting ovarian function, and regulating ovarian hormone secretion, and its efficacy is superior to that of cetrorelix or Zuogui pills alone.
ABSTRACT
High-intensity sound often leads to the dysfunction and impairment of central nervous system (CNS), but the underlying mechanism is unclear. The present study was aimed to investigate the related mechanisms of CNS lesions in Bama miniature pig model treated with high-intensity sound. The pigs with normal hearing were divided into control and high-intensity sound (900 Hz-142 dB SPL, 15 min) groups. After the treatment, hippocampi were collected immediately. Fluo-4 was used to indicate intracellular Caconcentration ([Ca]) change. Real-time PCR and Western blot were used to detect mRNA and protein expressions of calcium-sensing receptor, L-Cachannel α2/δ1 subunit, PKC and PI3K, respectively. DAPI staining was used to identify nuclear features. The result showed that high-intensity sound exposure resulted in significantly swollen cell nucleus and increased [Ca]in hippocampal cells. Compared with control group, high-intensity sound group showed increased levels of PI3K, PKC and L-Cachannel α2/δ1 subunit mRNA expressions, as well as up-regulated PKC and calcium-sensing receptor protein expressions. These results suggest that the high-intensity sound activates PKC signaling pathway and induces calcium overload, eventually leads to hippocampal injury, which would supply a novel strategy to prevent nervous system from high-intensity sound-induced injury.
ABSTRACT
Here we describe and illustrate our methods for multi-channel in vivo recording in mice, including the fabrication of the microdrive array and the surgical procedure for implanting electrodes. The multi-channel microdrive is fabricated from printed circuit board base, screws, nuts and clamping screws. Rotation of the screw drives both the nut and the attached electrodes to move forward simultaneously. Each full turn of the screw corresponds to 280 µm in depth penetration. The recording electrodes are self-made tetrodes consisting 4 wires (13 µm in diameter). The major steps of headstage fabrication include: tetrode making, microdrive construction, headstage assembling and tetrode plating. The finished headstage is suitable for multi-channel recording in freely moving rodents with the modest weight and the adjustable number of recording electrodes. Additionally, the recording site is allowed to be manipulated after implantation at any time. In the latter part of this paper, we introduce the procedure of the implant surgery to record in bilateral hippocampus in mice. Using these headstages, we simultaneously recorded population activity in bilateral CA1 in freely behaving mice.