Protective effect of quercetin on L-02 cells by inhibiting DNA damage of INH-induced mitochondrial oxidative stress / 中国病理生理杂志

AIM: To investigate the role of reactive oxygen species ( ROS)-mediated mitochondrial oxidative injury in isonicotinyl hydrazide ( INH)-induced DNA damage and the protective effect of quercetin on L-02 cells.ME-THODS:The injury model of hepatocyte L-02cells in vitro induced by INH was established .The cells were divided into control group, INH group, low-dose quercetin group and high-dose quercetin group.The DNA damage of L-02 cells was evaluated by the comet test .The mitochondrion was prepared , and the level of mitochondrial ROS and the value of mitochondrial membrane potential (ΔΨm ) were detected by fluorescent probes DCFH-DA and rhodamine 123.The content of MDA was measured by TBA method .The activity of SOD was assessed with the xanthine oxidase method .The protein expression of Bcl-2 and Bax was determined by Western blotting , and the value of Bax/Bcl-2 was calculated .RESULTS:INH induced obvious DNA damage , increased the level of mitochondrial ROS , the content of MDA and the value of Bax/Bcl-2, and markedly reduced the value of ΔΨm and the activity of SOD in the L-02 cells.Quercetin attenuated DNA dam-age, reduced the level of mitochondrial ROS , elevated the value of ΔΨm , declined the content of MDA , increased the ac-tivity of SOD and decreased the value of Bax/Bcl-2 in the L-02 cells.CONCLUSION:INH induces DNA damage in L-02 cells by generation of mitochondrial oxidative stress .Quercetin has a protective effect on L-02 cells to attenuate the INH-in-duced DNA damage by inhibiting ROS-mediated mitochondrial oxidative damage .

Oxygen free radical injury of myocardial mitochondria in the experimental type 2 diabetic rats / 中国病理生理杂志

AIM: To study the mechanism of diabetic cardiomyopathy and abnormality of oxygen free radicals. METHODS: The contents of myocardial cytosolic cytochrome C, mitochondria cytochrome C, mitochondrial calcium, NO, MDA and the activity of SOD and NOS were determined in diabetic rats induced by STZ. The pathological changes were observed under transmission electron microscope. RESULTS: Compared to the normal and ganoderma group, the levels of mitochondrial NO, iNOS, MDA, calcium and plasma Cyt-C in rat myocardium were higher (P<0.05), while mitochondrial Cyt-C and SOD were lowered in model group (P<0.05). The bouncary indistinct, disorganization, a focal loss of muscular fibril, myocardium mitochondria swelling, pulmonary vascular endothelial cellular swelling and obstructed lumen of the capillary were also observed under transmission electronic microscope. CONCLUSION: The findings indicate that oxyradical and lipid peroxidation might be associated with the damage of myocardial mitochondria in NIDDM rats. Cyt-C and mitochondrial calcium is also involved in the process.

Changes of mitochondria calcium and cytochrome C in epididymis associated with apoptosis in varicocele rats / 中国组织工程研究

BACKGROUND: Varicocele (VC) can induce the infertility in males, so the investigation on its mechanism is important for the treatment of male infertility. OBJECTIVE: To analyze the effects of VC induced by surgical operation on the contents of mitochondria calcium, cytochrome C and cell apoptosis as well as the changes of microstructure and ultrastructure in epididymis. DESIGN: Randomized control experiment. MATERIALS: The experiment was conducted in Jiamusi University between June 2003 and May 2004. Forty male adolescent Wistar rats with the average body mass of (220±20) g were selected, which were provided by the Animal Laboratory of Jiamusi University. Rats were randomly divided into sham-operation group and deligation group with 20 rats in each group at one week after feeding at room temperature. METHODS: Rats in the sham-operation group were made into sham-op eration models by exposing the left renal vein. Rats in the deligation group were deligated of partial left renal veins so as to establish VC models. Bilateral epididymides were removed at ten weeks after operation. The levels of mitochondria calcium in head and body of epididymis as well as the contents of cytochrome C and cytoplasm cytochrome C were detected. The cell apoptosis was detected by in situ terminal deoxynucleotityl transferase mediated dTUP nick end labeling (TUNEL) technique. The specimens of corpus epididymis were routinely made for observation under optimal microscope and electron microscope. The changes of microstructure and ultrastructure of epididymis were studied.MAIN OUTCOME MEASURES: The contents of mitochondria calcium, cytochrome C and cytoplasm cytochrome C. Cell apoptosis. Changes of cellular morphous in epididymis. RESULTS: A total of 40 rats were involved in the analysis of results.① The contents of mitochondria calcium in bilateral epididymis were obviously decreased in the deligation group than the sham-operation group [(4.72±1.45), (5.90±1.97), (10.13±2.34) mg/g, (P ＜ 0.01)].②The content of mitochondria cytochrome C in right epididymis obviously increased more in the deligation group than the sham-operation group [(0.36±0.20), (0.19±0.14), (0.15±0.07) μmol/L (P ＜ 0.05)]. ③The contents of cytoplasm cytochrome C in bilateral epididymis greatly increased more in the deligation group than the sham-operation group [(8.17±1.49), (7.48 ± 1.60), (5.93±1.60) mol/L, (P ＜ 0.05)].④The apoptotic rate of bilateral cells in the deligation group was significantly increased than the sham-op eration group [( 13.3±1.9)%, ( 12.6±1.5)%, (6.2±0.3)%,(P ＜ 0.01 )]. However, there were no significant differences in mitochondria calcium, cytochrome C, cytoplasm cytochrome C and apoptotic rate between the left and right mitochondria of the deligation group (P ＞ 0.05).⑤Main changes under light microscope: cuctus epididymis shrinked, the blebbing appeared in epithelial cells, and the light cells as well as halo cells in epithelia were significantly increased. ⑥Main representation under electron microscope: the cytolysosome inside the chief cells were increased and enlarged with increased residual bodies, and the endoplasmic reticulum expanded, the mitochondria cristae was dim, the Golgi complex was vacuolated. Besides, nuclear chromatin were dense and in lump at different size, which located mainly in the nuclear membrane. The microvilli of columnar epithelial cells were sparse and local defects could be seen. CONCLUSION: The cytochrome C is released to kytoplasm via mitochondrial outer membrane, which activates the caspase 3 and leads to the apoptosis, and accordingly causes excessive apoptosis of epididymal tissues and as well as the changes of microstructure and ultrastructure. All these changes may be one of the important reasons of infertility resulting from VC.

Study of varicocele and oxidative stress / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the mischief mechanism of oxidative stress in the varicocele (VC).</p><p><b>METHODS</b>Serum was taken from the spermatic and peripheral veins on ligation of the internal spermatic veins in 28 infertile males with VC. Experimental VC was established in male rats by partial ligation of the left renal vein. And testis tissue was taken three months after operation. The nitric oxide(NO), nitric oxide synthetase (NOS), xanthine oxidase (XO), lactic acid(Lac) and lactic dehydrogenase (LDH) in the serum of 28 infertile males with VC and the testis tissue of the VC rats were detected by spectrophotometry.</p><p><b>RESULTS</b>NO, NOS, XO and Lac in the serum of internal spermatic veins in the infertile males with VC were significantly higher than in the serum of peripheral veins in the VC patients (P < 0.01, P < 0.05). LDH was lower than that in peripheral serum. NO and XO of the left testis tissue in the VC rats were higher compared with the control group (P < 0.01). Lac in the left testis of the VC rats was lower than that in the control group rats (P < 0.01).</p><p><b>CONCLUSION</b>NO, NOS and XO in the serum of the VC patients and in the testis tissues of the VC rats were increased, and Lac and LDH were changed obviously, which might not only disturb spermatogenesis, but also inhibit sperm motility. Therefore they might be one of the causes of infertility in VC patients.</p>

Effects of experimental varicocele on mitochondria calcium and cytochrome C of the epididymis in adolescent rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of experimental varicocele on mitochondria calcium and cytochrome C of the epididymal cells in adolescent rats.</p><p><b>METHODS</b>Forty male adult Wistar rats were divided into two groups randomly: varicocele group (VG) and sham operation group (SOG) by partial ligation or exposure of the left renal vein. Bilateral epididymides were removed after ten</p><p><b>RESULTS</b>The content of mitochondria weeks. Mitochondria calcium and cytochrome C levels of the epididymal cells were detected. calcium decreased (P < 0.001 ) while that of cytochrome C increased (P < 0.05) markedly in the experimental group compared with SOG.</p><p><b>CONCLUSION</b>Calcium dyshomeostasis and mitochondrial damage of the epididymal cells caused by varicocele may play an important role in leading to subfertility.</p>

Study of the effect of super painkiller on the sexual function of male rats with type I diabetes / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the effects of super painkiller on the sexual function of male Wistar rats with diabetes.</p><p><b>METHODS</b>Thirty-two Wistar rats at 3-5 months of age were divided into two groups at random: 22 in the diabetes group, and 10 in the control group. After 72 hours, the former was further divided into 2 subgroups: non-treatment and super painkiller treatment. In 5 weeks, blood glucose was determined. After the apomorphines experiment, the rats were killed, blood taken from the vein, homogenate prepared from the isolated testis tissue and the level of NO and NOS in the serum and tissue homogenate surveyed, separately.</p><p><b>RESULTS</b>Compared with the control group, serum NO and NOS of the diabetic rats dropped sharply. Compared with the non-treatment group, the serum NO and NOS of the super painkiller treatment group were very high and the difference was significant (P < 0.01). Apomorphine injection showed that the times of penis erection of the treated rats were more than those of the non-treatment group (P < 0.01) , but the difference was not significant compared with the control (P > 0.05).</p><p><b>CONCLUSION</b>Super painkiller has sure effect of reducing blood glucose, with a certain curative value for sexual and reproductive malfunction of diabetic rats.</p>

Effects of oleanolic acid on apoptosis and cell cycle of HL-60 cells in vitro / 中国病理生理杂志

AIM:To study the mechanism of oleanolic acid induced apoptosis and its influence on cell cycle in HL-60 cells in vitro.METHODS:The HL-60 cells were treated with different concentrations of oleanolic acid and then cultured for 12 h,24 h,48 h and 72 h,respectively.MTT assay was used to evaluate the inhibitory effect of oleanolic acid on HL-60 cells in vitro.The argarose gel electrophoresis was used to detect the chromatin DNA fragmentation.FACS was used to analyze the cell cycle of HL-60 cells.Western blotting was used to detect the activation of caspase-3 which has been confirmed the last execution of apoptosis pathway.RESULTS:MTT assay showed that oleanolic acid dramatically inhibited the growth of HL-60 cells in vitro,more than 50% HL-60 cells were inhibited when the cells were treated with 80 ?mol/L oleanolic acid for 48 h;the apparent DNA ladder was detected after exposure of HL-60 cells to oleanolic acid for 48 h.FACS analysis showed that cell cycle of HL-60 cells was arrested in G1 phase,the inhibition ratio of HL-60 cells achieved 63.24% and 67.90% after treated with oleanolic acid for 48 h and 72 h correspondingly.Western blotting detected the activation of caspase-3 after exposure of HL-60 cells to 80 ?mol/L oleanolic acid for 48 h.CONCLUSION:These results suggest that oleanolic acid induces apoptosis and the cell cycle of HL-60 cells is arrested in G1 phase.

Effects of hypothermia on the contents of neuropetides and other active substances in rat plasma and different brain regions / 中国病理生理杂志

AIM: To study the effects of hypothermia on the contents of AVP,Ang Ⅱ,L-EK,cAMP,Mg 2+ and Ca 2+ in rat plasma and different brain regions.METHODS: The model of experimental hypothermia rat was established and treated in groups randomly.At the end of the experiment,samples of plasma and braintissues were taken and the contents of L-EK,cAMP,AVP,Ang Ⅱ were determined by RIA methods,Mg 2+ and Ca 2+ by atom absorption methods.RESULTS: The contents of Ang Ⅱ,AVP increased significantly in the plasma of Ⅰ,Ⅲ group( P

Effect of Ganoderma lucidum spores on IL-1? and c-Fos in the brain tissue of epilepsy rats / 中国病理生理杂志

AIM: To observe the influence of Ganoderma lucidum spores on the cytokine IL-1? and c-Fos in the brain tissue of epilepsy rats. METHODS: The IL-1? level was examined by radioimmunoassay and the c-Fos expression was measured by immunohistochemical assay. RESULTS: Comparing the Ganoderma lucidum spores group with the epilepsy model group: c-Fos positive cell count in brain cortex and hippocampus in treatment group was obviously reduced. IL-1? level in brain tissue was also reduced obviously. CONCLUSION: Ganoderma lucidum spores effectively reduces cytokine IL-1? in brain tissue of epilepsy rats, improves the immunity dysfunction and plays a role in anti-epilepsy through suppressing c-Fos expression in epilepsy rats brain tissue and blocking of LRG.

Effect of Ganoderma lucidum spores powder on the expression of IGF-1, NF-?B and apoptosis of nerve cells in the brain from epileptic rat / 中国病理生理杂志

AIM: To investigate the effect of Ganoderma lucidum spores powder on the expression of insulin-like growth factor-1 (IGF-1), nuclear factor-?B (NF-?B) and apoptosis of nerve cells in rats with epilepsy established by pentetrazole. METHODS: The sub-eclampsia dosage of pentetrazole (PTZ) was used to make epilepsy model. Ganoderma lucidum spores powder group was given from stomach. The enduring time and latent period were recorded. The immune reactivity of IGF-1, NF-?B/P65 and apoptosis of nerve cells were measured with immunohistochemical staining and TUNEL method. RESULTS: In high power sight (?400), there were much more apoptosis cells in hippocampus and brain cortex of model group (18.80?2.13, 16.87?2.00) than those in control group (0.97?0.52, 0.58?0.25). The expressions of IGF-1, NF-?B in model group were higher than those in control group. Compared with model group, the latent period of Ganoderma lucidum spores powder group at the 17th, 21th, 25th days were longer (P

Study of experimental varicocele on cell apoptosis of epididymis in adolescent rats / 中国病理生理杂志

AIM: To study the cell apoptosis and the change of microstructure and ultrastructure in epididymis with experimental varicocele (EVC) in rats. METHODS: Experimental varicocele model was induced by partial ligation of the left renal vein in adolescent Sprague-Dawley Wistar rats. Apoptotic cells were detected by in situ terminal deoxynucleotityl transferase-mediated dTUP nick end labeling (TUNEL) technique. The corpus epididymis of the rats was prepared for light and electron microscopic observation. The microstructure and ultrastructure of the epididymis were studied. RESULTS: There was certain proportion of apoptosis cells in epididymis cells in control rats. The incidence of apoptosis increased remarkably in experimental group than that in control group (P

Effects of thymosin ?_1 on the development and maturation of thymocytes / 中国病理生理杂志

AIM: To investigate the effect of Thymosin ? 1 on the development and matutation of thymocytes. METHODS: The proportion of CD4+CD8+ thymocytes and the expression of smoothened (Smo) of the hedgehog (Hh)-signaling in CD4-CD8-thymocytes were examined to observe the effect of thymosin ? 1 on thymocyte development and matutation. RESULTS: Flowcytometric analysis showed that thymosin ? 1 showed activity at a low dose of 30 ?g/kg, and 30 ?g/kg thymosin ? 1 accelerated the replenishment and maturation of thymocytes according to the expression of Smo of the Hh-signaling in CD4-CD8-thymocytes, the potent negative regulator of proliferative responses. CONCLUSION: Thymosin ? 1 can accelerate thymocyte development from CD4-CD8- to CD4+CD8+.

Study on the mitochondria Ca 2+, expression of Bcl-2 and Bax and germ cell apoptosis in testes of rats with varicocele / 中国病理生理杂志

AIM: The cytoplasm Ca 2+, expression of Bcl-2 and Bax and germ cell apoptosis of testes in adult male rats with varicocele were observed to investigate the relationship between apoptosis and infertility. METHODS: Thirty-five healthy male Wistar rats were divided into two groups randomly: varicocele group (VG, n=20) and sham operation group (SOG, n=15). Bilateral testes were removed after ten weeks. One part of it was homogenized, the other part was performed for tissue sectioning. The atomic absorption method was used to detect the cytoplasm Ca 2+. The germ cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling (TUNEL) technique. The expression of Bcl-2/Bax was detected by immunohistochemical method. RESULTS: The cytoplasm Ca 2+ in testes of rats with varicocele was decreased obviously. The apoptosis indexes of germ cells and Bax expression in VG were significantly increased as compared with those in the SOG, while Bcl-2 expression was obviously decreased. CONCLUSION: The results showed that apoptosis was increased in testes of VG rats, suggesting that male infertility with varicocele may be caused by some apoptosis agents such as high temperature, testes toxin and oxygen free radical. The cytoplasm Ca 2+ and change of expression of Bcl-2/Bax may contribute to germ cell apoptosis and cause male infertility.

Effect of ischemic preconditioning on contents of cytochrome C and mitochondrial calcium in rats after focal cerebral ischemic-reperfusion / 中国病理生理杂志

AIM: To observe the effect of ischemic preconditioning on contents of cytochrome C and mitochondrial calcium in rats after focal cerebral ischemic-reperfusion. METHODS: Focal cerebral ischemic model was made by occlusion of right middle artery in Wistar rats (ischemia for 2 h and reperfusion for 4 h). Rats were randomly divided into three groups: ischemia pretreatment, model and sham operation. Rats in ischemic pretreatment group were undergone transient ischemic preconditioning (30 min) and reperfusion (72 h). The contents of cytochrome C were measured according to Zhangjuntian's improved methods. The contents of mitochondrial calcium were detected by flame atom absorption. RESULTS: The contents of mitochondrial cytochrome C and calcium in model group were significantly lower than those in sham operation (P