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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.
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AIM: To explore the role and mechanism of silent mating type information regulator 2 homolog 3 (SIRT3) in attenuation of intestinal ischemia-reperfusion (I/R) injury by dexmedetomidine in mice. METHODS: Twenty-four healthy male C57BL mice were divided into 4 groups randomly (n=6): sham operation group (Sham group), intestinal ischemia-reperfusion group (I/R group), dexmedetomidine group (Dex group), SIRT3 inhibitor 3-TYP group (3-TYP group). Superior mesenteric artery was clamped for 45 min followed by reperfusion for 2 h to establish intestinal I/R model in I/R group, Dex group, and 3-TYP group. Sham group received sole sham operation. 1 h prior to onset of ischemia, 3-TYP was injected into mice in 3-TYP group intraperitoneally (5 mg/kg, diluted to 0.3 mL), and 0.3 mL normal saline into mice in Dex group intraperitoneally. 30 min prior to onset of ischemia, dexmedetomidine was injected into mice in 3-TYP group and Dex group intraperitoneally (25 μg/kg, diluted to 0.3 mL). 1 h and 30 min prior to onset of ischemia, 0.3 mL normal saline was injected into mice in Sham group and I/R group intraperitoneally, respectively. 2 h of after reperfusion, the mice were sacrificed under anesthesia. Intestinal tissues were took and observed for pathological changes under light microscope after HE staining, and the injury was assessed via the Chiu's score method, and activities of SIRT3 and superoxide dismutase 2 (SOD2) were detected via spectrophotometry, and malondialdehyde (MDA) via spectrophotometry. RESULTS: The pathological injury was exacerbated, and the Chiu's score, the MDA level elevated remarkably, while the activity level of SIRT3 and SOD2 declined remarkably in I/R group, Dex group and 3-TYP group compared to Sham group (P<0.05). The pathological injury was alleviated, and the Chiu's score declined remarkably in Dex group and 3-TYP group compared to I/R group (P<0.05); and the MDA level declined remarkably, while activity level of SIRT3 and SOD2 elevated remarkably in Dex group compared to I/R group (P<0.05); and there was no significant difference both in the activity level of SIRT3 and SOD2 and in the MDA level between 3-TYP group and I/R group. The pathological injury was exacerbated, and the Chiu's score, the MDA level elevated remarkably, while the activity level of SIRT3 and SOD2 declined remarkably in 3-TYP group compared to Dex group (P<0.05). CONCLUSION: SIRT3 and its downstream SOD2 are involved in mediating the effect of attenuation of intestinal ischemia-reperfusion injury through inhibiting oxidative stress response by dexmedetomidine.
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@#AIM:To investigate the protective effects of resveratrol(RSV)on inflammation and oxidative stress damage in human corneal epithelial cells(HCECs).<p>METHODS: The inflammation of HCECs was induced by Tumor necrosis factor-α(TNF-α), and the experiment was divided into: control group, TNF-α group and RSV+TNF-α group. The oxidative stress response of HCECs was induced by H<sub>2</sub>O<sub>2</sub>, and they were divided into normal group, H<sub>2</sub>O<sub>2</sub> group and RSV+ H<sub>2</sub>O<sub>2</sub> group. MTT assay was used to detect the viability of HCECs; RT-qPCR and enzyme-linked immunosorbent assay(ELISA)methods were used to detect the expression of IL-1, IL-6 and IL-8; Immunofluorescence staining and Western blot were used to observe the nuclear translocation of NF-κB p65. 2',7'-dichlorofluorescein diacetate(DCFH-DA)fluorescent probe was applied to detect the level of reactive oxygen species(ROS).<p>RESULTS:In the inflammatory response of HCECs, RT-qPCR and ELISA showed that the expression levels of IL-1, IL-6 and IL-8 were increased significantly in the TNF-α group compared with the control group, the above indicators were lower after pretreatment of RSV than those in TNF-α group; Immunofluorescence staining and Western blot showed that the nuclear translocation of NF-κB p65 was increased in TNF-α group, while it was inhibited after pretreatment of RSV. In the oxidative stress response of HCECs, the results of MTT and DCFH-DA fluorescent probe staining showed that H<sub>2</sub>O<sub>2 </sub>significantly decreased the viability of HCECs and increased the production of ROS in HCECs. After pretreatment of RSV, cell viability increased significantly, and RSV inhibited the generation of ROS in HCECs induced by H<sub>2</sub>O<sub>2</sub>. <p>CONCLUSION: RSV has an inhibitory effect on inflammation and oxidative stress damage in human corneal epithelial cells, and it has been confirmed that RSV inhibits inflammation by inhibiting the activation of the NF-κB pathway.
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Objective To investigate the role of histone deacetylase 3 ( HDAC3 ) inhibitor (HDAC3I) in hypoxia-reoxygenation( H/R) injury of PC12 cells. Methods H/R cell injury model was established by using PC 12 cells for 4 hours hypoxia and then reoxygenation for 24 hours. HDAC3I treatment group was pretreated with RGFP966 for 1 hour and then subjected to hypoxia-reoxygenation injury. The experiment was divided into three groups: normal control group, model group and HDAC3I treatment group, and 3 repetitions for each group. Cell viability was determined using MTT. Cellulose dehydrogenase (LDH) was detected by colorimetry. Flow cytometry was used to detect apoptosis and intracellular reactive oxygen species ( ROS ), respectively. The activity of superoxide dismutase ( SOD ) was determined by xanthine oxidase method. The malondialdehyde ( MDA ) content was determined by the thiobarbituric acid method. Western blotting was used to detect the expression of Bax, Bcl-2, cleaved-Caspase-3 and HDAC3 proteins. Results Compared with the control group, the cell viability of the model group and HDAC3I treatment group decreased significantly (P<0. 05), and the cell LDH (P<0. 05) and apoptosis (P<0. 05) increased significantly. The cell viability of HDAC3I treatment group was significantly higher than that of the model group (P<0. 05), while the LDH (P<0. 05) and apoptosis of HDAC3I treatment group were lower than the model group (P<0. 05). In addition, compared with the control group, the ROS and MDA (P< 0. 05 ) of the model group and the HDAC3I treatment group increased significantly, and the SOD decreased significantly (P<0. 05). ROS and MDA in the HDAC3I treatment group (P<0. 05) were significantly lower than the model group, while the SOD level was higher than the model group (P< 0. 05). Western blotting analysis showed that compared with the control group, Bax and cleaved-Caspase-3 in the model group and HDAC3I treatment group increased significantly, and Bcl-2 decreased significantly (P<0. 05). The Bax and cleaved-Caspase-3 in the HDAC3I treatment group were significantly lower than the model group, and Bcl-2 was significantly higher than the model group (P<0. 05). Compared with the control group, the expression of HDAC3 protein in the model group increased significantly (P<0. 05), while the HDAC3 protein in the HDAC3I treatment group decreased significantly (P<0. 05). Conclusion HDAC3I reduces PC 12 cell apoptosis induced by hypoxia/reoxygenation by reducing oxidative stress.
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AIM: To investigate the effect and mechanism of hypoxia inducible factor-1α on intestinal mucosal barrier in sepsis. METHODS: SD rats were randomly divided into 4 groups: sham group, sepsis group, sepsis+HIF-1α stimulant (sepsis+DMOG group), sepsis+HIF-1α inhibitor (sepsis+Bay87-2243 group), 6 rats in each group. Sepsis model was established by cecal ligation and perforation (CLP). The levels of inflammatory markers IL-1β, IL-6, TNF-α, oxidative stress markers MDA and antioxidant factors SOD and CAT were detected by ELISA and the expression of HIF-1α in intestinal mucosa was detected by Western blot. The pathological damage of intestinal mucosa was detected by HE staining. RESULTS: Inflammatory factors, oxidative stress factors and HIF-1α were significantly up-regulated in septic rats (P<0.05). The contents of IL-1β, IL-6, TNF-α and MDA in plasma were significantly decreased by intraperitoneal injection of DMOG (P<0.05); the levels of SOD and CAT in plasma were increased (P<0.05), HIF-1α was up-regulated (P<0.05), and the pathological damage of intestinal mucosa was alleviated, with decreased Chiu's score (P<0.05). Oral administration of Bay87-2243 gave the opposite result. CONCLUSION: HIF-1α has a protective effect on intestinal mucosal injury in sepsis. The mechanism may be related to the alleviation of inflammatory response and inhibition of oxidative stress.
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Objective: To investigate the effect of vitexin on oxidative stress in rats with acute cerebral ischemia-reperfusion by regulating the pathway of nuclear factor E2-related factor 2 (Nrf2)/antioxidant response element (ARE). Methods: A total of 54 SD rats were randomly divided into Sham group, model group, positive control (edaravone 0.56 mg/kg) group and vitexin low, medium, and high dose (10, 20, 40 mg/kg) groups. The rat models with acute cerebral ischemia were established except the Sham group. After reperfusion, rats in Sham group and model group were received ip saline. The edaravone group and vitexin groups were administered according to the corresponding dose, once every 8 h for a total of three times. The neurobehavioral scores of rats before and after intervention were compared. HE staining was used to detect the pathological changes of cerebral cortex. The levels of MDA, NO, SOD, and GSH in cerebral cortex of rats were detected by the kit. The mRNA and protein expression levels of Nrf2/ARE pathway related gene in rat cerebral cortex were detected by real-time fluorescent quantitative PCR (qRT-PCR) and Western blotting. Results: Before and after intervention, there was no significant change in the neurobehavioral score of rats in the Sham group, the model group was higher than before intervention (P < 0.01), the edaravone group and the vitexin groups were lower than before intervention (P < 0.01), and there was a significant difference in the neurobehavioral score between the groups after intervention (P < 0.01). In the Sham group, the distribution of nerve cells was uniform and dense, and the morphology of cell body and nucleus was normal. In the model group, edaravone group and vitexin group, the arrangement of brain tissue was disordered and loose, in which liquefying necrosis and disappearance of cell structure were observed in the model group; in the low-dose group, the arrangement of cells was seriously disordered and loose. In the vitexin medium dose group and edaravone group, the area of liquefying necrosis was small, and most of the cells were normal; In the vitexin high dose group, only a few liquefying necrosis were found, the cell structure was basically normal, and the cell arrangement was slightly disordered. Compared with the Sham group, the levels of MDA and NO in the cerebral cortex of the model group were significantly increased (P < 0.01), the levels of SOD and GSH were significantly reduced (P < 0.01), and the expression levels of Nrf2 and γ-GCS mRNA were significantly increased (P < 0.01), the expression of cytoplasmic Nrf2 and γ-GCS proteins were significantly increased (P < 0.01), and the expression levels of nuclear Nrf2 and HO-1 proteins were significantly decreased (P < 0.01). Compared with the model group, the levels of MDA and NO in the cerebral cortex of rats in the edaravone group and low, medium and high dose groups of vitexin were significantly reduced (P < 0.01), and the levels of SOD and GSH were significantly increased (P < 0.01), Nrf2 and γ-GCS mRNA expression levels were significantly reduced (P < 0.01), cytoplasmic Nrf2, γ-GCS protein expressions were significantly reduced (P < 0.01), and nuclear Nrf2 and HO-1 protein expression levels were significantly increased (P < 0.01). And the levels of MDA, NO, SOD, GSH and Nrf2, γ-GCS, HO-1 mRNA and protein expression in rat cerebral cortex in each dose group were dose-dependent, with significant differences between groups (P < 0.01). Conclusion: Vitexin can alleviate oxidative stress in rats with acute cerebral ischemia-reperfusion. It is speculated that it is related to the regulation of Nrf2/ARE signaling pathway, the up-regulation of Nrf2 gene and protein expression, the promotion of its movement from cytoplasm to nucleus, the up-regulation of HO-1 expression, the inhibition of γ-GCS expression, and the enhancement of the body’s ability to response to oxidative stress.
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Objective: To investigate the therapeutic effects of YiXuanNing Granule on the ischemic vertigo of the mice and rats, and to clarify its effects on the inflammation damage and oxidative stress in brain tissue of the mice and rats Methods: The vertigo models of mice and rats were established by mechanical vertigo and ligation of bilateral common carotid arteries (CCA), respectively. A total of 60 mice and 60 rats were selected and randomly divided into normal control group, model group, low dose of YiXuanNing Granule group (2.25 g middot; kg-1/1. 50 g middot; kg-1), middle dose of YiXuanNing Granule group (4.50 g middot; kg-1/3. 00 g middot; kg-1), high dose of YiXuanNing Granule group (9.00 g middot; kg-1/6. 00 g middot; kg-1) and positive control group (1.50 g middot; kg-1/1. 00 mg middot; kg -1 flunarizine hydrochloride capsule), and there were 10 animals in each group. The mice and the rats in control group and model group were intragastrically administered with 10 mL middot; kg-1 normal saline, and the mice and the rats in the other groups were given 10 mL middot; kg: corresponding drug once a day for 7 d. The escape time of jumping off the platform of the mice and the rats in various groups were determined; the food intake of the mice in various groups were detected; the activities of superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) in brain tissue of the rats in various groups were detected by colorimetric method. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in brain tissue of the rats in various groups were detected by ELISA method. Results: Compared with model group, the escape time of jumping off the platform of the mice and the rats in different doses of YiXuanNing Granule groups and positive control group was shortened (P<0. 05 or P<0. 01), and the food intake of the mice in various groups was increased. Compared with model group, the SOD activities in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly increased (P<0. 05 or P<0. 01); the MDA levels in brain tissue of the rats in different doses of YiXuanNing Granule groups were significantly decreased (P<0. 01); compared with model group, the levels of TNF-α and IL-1β in brain tissue of the rats and the mice in middle and high doses of YiXuanNing Granule groups were significantly decreased (P<0. 05 or P<0. 01). Conclusion: YiXuanNing Granule can inhibit the inflammatory damage in brain tissue of the rats with ischemic vertigo and reduce the oxidative stress, thereby, it can play a therapeutic role in ischemic vertigo.
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OBJECTIVE: To investigate the effect of transcutaneous electrical acupoint stimulation (TEAS) on pulmonary function, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content in patients using tourniquet after lower extremity surgery. METHODS: A total of 40 patients who underwent lower extremity surgery were equally randomized into control group and TEAS group by using a random number table. All patients underwent lumbar epidural anesthesia combined with block anesthesia. The patients in the TEAS group were given TEAS at Zusanli (ST36) and Sanyinjiao (SP6) beginning from 30 min before surgery to the end of surgery, and those in the control group received TEAS at the same acupoints with minimum current intensity. Mean arterial pressure (MAP) and heart rate (HR) were recorded at each time point (T0: pre-surgery /TEAS, T1: 5 min after anesthesia, T2: 1 min before tourniquet-loosening, T3: 1 min after tourniquet-loosening, T4: 5 min after tourniquet-loosening, and T5: 6 h after tourniquet-loosening). Blood samples (4 mL) was collected from the radial artery before TEAS and 6 h after loosening tourniquet for analyzing blood gas parameters as partial pressure of caron dioxide(PCO2), arterial partial pressure of oxygen (PaO2), alveolar partial pressure of oxygen (PAO2), alveolar-arterial oxygen pressure difference (PA-aDO2) and respiratory index (RI) by using a blood gas analyzer, and plasma SOD activity and MDA content were assayed by using xanthine oxidase method and thiobarbituric acid colorimetry method, respectively. RESULTS: Intra-group comparison showed that compared with T0, a significant increase was found in PA-aDO2 and RI at T5 and a significant reduction in PaO2 and PaO2/ PAO2 (a/A) ratio in the control group (P<0.05), and the same changes in the TEAS group (P<0.05) except a/A ratio. Comparison between two groups showed that at T5, both PaO2 and a/A levels were significantly higher in the TEAS group than in the control group (P<0.05), and both PA-aDO2 and RI levels were obviously lower in the TEAS group than in the control group (P<0.05), suggesting an improvement of the pulmonary function after TEAS. At T5, plasma SOD activity was significantly decreased and plasma MDA content was remarkably increased in the control group relevant to T0 (P<0.05), SOD activity was significantly higher in the TEAS group than in the control group (P<0.05), and MDA content was evidently lower in the TEAS group than in the control group (P<0.05), suggesting a reduction of oxidative stress response after TEAS. CONCLUSION: TEAS at ST36 and SP6 can improve pulmonary function and attenuate oxidative stress response in patients using tourniquet after lower extremity surgery.
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Objective: To study the therapeutic effect of point injection of Salvia Miltiorrhiza and Ligustrazine Hydrochloride Injection (SMLHI) on diabetic peripheral neuropathy (DPN) patients and its effect on oxidative stress response. Methods: A total of 100 DPN patients were selected from the Neurology Department of the Fifth Affiliated Hospital of Zhengzhou University from March 2016 to February 2018. According to the random number table method, the patients were divided into observation group and control group with 50 cases in each group. Each group was treated with routine hypoglycemic drugs, diet control and exercise control, while the control group was treated with Mecobalamin Tablets. The observation group was treated with Mecobalamin Tablets combined with SMLHI at acupoints for 4 weeks as a course of treatment for two consecutive courses of treatment. The clinical symptom score, serum superoxide dismutase (SOD), gamma-glutamyltransferase (GGT), ferritin (SF), total anti-oxidant capacity (T-AOC), and malondialdehyde (MDA) levels, median nerve and peroneal nerve conduction velocity (MNCV), median nerve, peroneal nerve conduction velocity (SNCV), and curative effect were observed and compared before and after treatment. Results: After treatment, the clinical symptom score of the observation group was significantly lower, and the total effective rate of the observation group was 84.00%, which was higher than that of the control group 64.00% (P < 0.05). The SOD level in the observation group was higher, while the GGT, SF, T-AOC, and MDA levels were lower (P < 0.05). The MNCV and SNCV in the observation group were higher (P < 0.05). Conclusion: Acupoint injection of SMLHI for DPN patients can significantly reduce oxidative stress reaction in vivo, promote the recovery of nerve function, improve clinical symptoms of patients with high safety, which is worthy of clinical promotion.
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Objective To study the effects of exchange transfusion on renal function, inflammatory factors and blood internal environment in children with hyperbilirubinemia. Methods Neonates with hyperbilirubinemia treated in neonatal intensive care unit of the hospital between February 2015 and March 2017 were selected as the research subjects and divided into two groups by random number table method. Combined group received exchange transfusion combined with blue ray irradiation, and control group received blue ray irradiation. The serum levels of bilirubin, renal function indexes, inflammatory response indexes and oxidative stress response indexes were measured 24 hours after the treatment. Results 24 hours after the treatment,serum TBIL,BUN,Cr,β2-MG, CysC, TRF, CRP, IL-6, TNF-α, MDA and AOPP levels of combined group were greatly lower than those of control group whereas SOD, CAT and GSH-Px contents were greatly higher than those of control group. Conclusion Exchange transfusion can improve the renal function and inhibit the inflammatory and oxidative stress response in children with hyperbilirubinemia.
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Objective To investigate the protective effect and possible mechanism of Gossypol on isolated myocardial ischemia/reperfusion injury in rats.Methods 40 male Sprague-Dawley rats were randomly divided into 4 groups:Blank group,heart ischemia reperfusion group (MI/R group),high and low-dose Gossypol group (40,20 mg/L).The model of the myocardial ischemia/reperfusion injury were established using the Langendorff method.The hemodynamicsindexes,cardiac enzymes AST and LDH,inflammatory cytokines (NF-κB,ICAM-1,TNF-α and IL-6) were measured.The effect and mechanism of Gossypol on early-stage MI/R of the oxidative stress response and the JNK/p38 MAPK signal pathway were investigated.Results Experimental results showed that Gossypol could significantly improve the functional capacity of the heart,reduce the contents of AST,LDH and inflammatory cytokines in reperfused heart tissue,and increase superoxide dismutase levels to protect the heart.The mechanism of this substance may involve anti-lipid peroxidation and inhibition of p38 kinase phosphorylation and JNK,and reduction of oxidative stress injury and apoptosis damage induced by MI/R.Conclusion This study confirm that Gossypol exerts extensive anti-MI/R effects.Its mechanism may be related to the interfering with the oxidative stress response and suppressing the JNK/p38 MAPK signal pathway.
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OBJECTIVE: To investigate the protective effect of mangiferin on acute spinal cord injury (SCI) in rats and its mechanism.