Effect of hydrogen molecule on apoptosis-related proteins and Nrf2 signa-ling pathway in glomerular mesangial cells cultured with high glucose / 中国病理生理杂志

AIM:To investigate the role of hydrogen molecule on apoptosis-related proteins in glomerular me-sangial cells cultured with high glucose and to explore its possible mechanism.METHODS:Mouse glomerular mesangial cells cultured in vitro were divided into 4 groups:normal control group(C group,5.5 mmol/L glucose),mannitol group(G group,5.5 mmol/L glucose+19.5 mmol/L mannitol),high glucose group(H group,25 mmol/L glucose),high glu-cose+hydrogen-rich water group(HH group,25 mmol/L glucose+hydrogen-rich water),and cultured for 48 h.The pro-tein levels of Bax,Bcl-2,cleaved caspase-3,nuclear factor E2-related factor-2(Nrf2),heme oxygenase-1(HO-1)and NAD(P)H:quinone oxidoreductase-1(NQO-1)were determined by Western blot ,and the mRNA expression of HO-1 and NQO-1 was determined by RT-PCR.The level of intracellular reactive oxygen species(ROS)was detected by dihydro-ethidium method,and the activity of superoxide dismutase(SOD)was measured by WST-8 assay.RESULTS:Compared with C group,the protein levels of Bax and cleaved caspase-3 were up-regulated,and Bcl-2 was down-regulated in H group(P <0.05).No significantly difference of the protein levels mentioned above between C and HH group was observed. Compared with H group,the protein levels of Bax and cleaved caspase-3 were down-regulated,and Bcl-2 was up-regulated in HH group(P <0.05).The level of intracellular ROS was higher and the activity of SOD was lower in H group than those in C group(P<0.05).However,there was no difference of the SOD activity between C group and HH group.The level of intracellular ROS decreased and the activity of SOD increased in HH group as compared with H group(P<0.05). Compared with C group,clearly reduced protein expression of Nrf2,HO-1 and NQO-1,and decreased mRNA expression of HO-1 and NQO-1 in H group were observed(P<0.05).Compared with H group,the protein levels of Nrf2,HO-1 and NQO-1 as well as the mRNA levels of HO-1 and NQO-1 were obviously increased in HH group(P<0.05 ).CONCLU-SION:Hydrogen molecule inhibits the expression of pro-apoptotic proteins and induces the expression of anti-apoptotic pro-teins in glomerular mesangial cells cultured with high glucose.The mechanism may be related to activation of Nrf 2 signaling pathway.

Progress on surgical treatment for femoral head-preservering in the precollapse stage of femoral head necrosis / 中国骨伤

Osteonecrosis of the femoral head(ONFH), a refractory disease characterized by death of the osteocytes and the bone marrow due to inadequate blood supply caused by various mechanisms, usually leads to the collapse of the femoral head and malfunction of the hip joint. The crux is to diagnose ONFH early in the precollapse stage and prevent subsequent progression of collapsing through early interventions, thus delaying or avoiding the replacement of the hip joint. A number of joint salvaging operation treatments for early stage ONFH are available. However, there has been no consensus with regard to the ideal treatment. The main trend now is to unite core decompression with bone-grafting, tantalum rod, bone marrow mesenchymal stem cell (BMSC) and other treatments. Also there are ways of osteotomy altering the angle of the femoral neck to relocate necrotic tissue from the weight-bearing segment. The implanting of tantalum rod remains controversial and the advent of bone marrow mesenchymal stem cell (BMSC) holds huge potential.

Expression of steroidogenic enzymes in the rat model of polycystic ovary syndrome / 生理学报

The aim of the present study was to investigate the expression changes of three steroidogenic enzymes in the polycystic ovary syndrome (PCOS). Thirty Sprague-Dawley (SD) rats were randomly divided into normal control (NC) group and PCOS group. PCOS rat model was established by DHEA injection. The serum levels of progesterone, estrogen and testosterone were measured by immunoradioassay or enzyme immunoassay. The cellular distributions of 3β-hydroxy steroid dehydrogenase (3β-HSD), 17β-hydroxy steroid dehydrogenase (17β-HSD) and cytochrome P450 aromatase (P450arom) in ovaries were detected by immunohistochemistry. The expression levels of 3β-HSD, 17β-HSD and P450arom were detected by RT-PCR and Western blot. The results showed that the serum levels of estrogen and testosterone of PCOS group were significantly higher than those of the NC group. There was no significant difference of serum progesterone level between the PCOS and NC groups. Compared with the NC group, the PCOS group showed increased mRNA and protein expressions of both 3β-HSD and 17β-HSD, as well as reduced P450arom mRNA and protein expressions. These results suggest that 3β-HSD and 17β-HSD, but not P450arom, may participate in the ovarian hormonal regulation in the present rat model of PCOS.

The effects of ACEI on calpain-mediated cardiomyocytes apoptosis and cardiac function in diabetic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of angiotensin converting enzyme inhibitor (ACEI) captopril on Calpain-mediated cardiomyocytes apoptosis and cardiac function in diabetic rats.</p><p><b>METHODS</b>Thirty adult male SD rats were randomly divided into 3 groups (n = 10), normal control group (NC group), diabetes mellitus group (DM group)and captopril treated group (Cap group). Streptozocin (STZ) were used to make the model of diabetes mellitus, captopril was administrated by gavage at the dose of 50 mg/kg every day, while in NC group and DM group the same volume of normal saline was administrated. Twelve weeks later, left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVDEP), maximal rise rate of left ventricular pressure (+ dp/dtmax) and maximal fall rate of left ventricular pressure (- dp/dtmax) were detected; Western blot was used to detect the expression of Calpain-1 Calpain-2, Bcl-2, Bax and total Caspase3 protein; apoptosis index (AI) were assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL).</p><p><b>RESULTS</b>Compared with NC group, LVDEP was significantly higher; LVSP, + dp/dtmax and - dp/dtmax were significantly decreased (P < 0.05); Bcl-2 protein expression was decreased; the expression of Calpain-1, Calpain-2, Bax and total Caspase3 protein were increased; the value of AI was significantly increased. Compared with DM group, LVDEP was significantly lower; LVSP, + dp/dtmax and - dp/dtmax were significantly increased (P < 0.05); Bcl-2 protein expression was increased, the expression of Calpain-1, Calpain-2, Bax and total Caspase3 protein were decreased; the value of AI was significantly decreased (P < 0.05).</p><p><b>CONCLUSION</b>Captopril can protect diabetic myocardial structure through inhibiting activation of Calpain-1 and Calpain-2, up-regulating the expression of Bcl-2, down-regulating the expression of Bax to inhibit Caspase3 dependent apoptosis, thereby improving the ventricular function and myocardial structure.</p>

Effects of extract of ginkgo biloba on learning and memory ability and NGF and NT-3 expression in diabetic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of extract of Ginkgo Biloba(EGB) on nerve growth factor(NGF) and Neurotrophin-3(NT-3) expression of hippocampus neurons in streptozotocin-induced type I diabetic rats.</p><p><b>METHODS</b>Thirty male SD rats were divided into three groups (n = 10): the control group, diabetic group and EGB-treated group. Strepozotocin were injected intraperitoneally in the later two groups to induce diabetes. EGB-treated group was injected intraperitoneally with EGB, and the same volume of normal saline was injected to the other groups. Concentration of blood glucose and body weight and behaviour were dynamicly monitored. At the end of the 12th week, morphological changes of the hippocampus neurons were observed under microscopy by HE stain. The expression of NGF and NT-3 were assayed by Western blot and RT-PCR respectively.</p><p><b>RESULTS</b>Compared with diabetic group, the behaviour and body weight (P < 0.05) and the concentration of blood glucose (P < 0.05) were significantly improved and the escape latency of Morris water maze test (P < 0.05) was significantly shortened, while the platform searching score was significantly increased (P < 0.01) in EGB treated group; The pathological changes of hippocampus neurons were significantly attenuate by EGB treated; The expression of NGF and NT-3 in hippocampus neurons were significantly increased which assayed by Western blotting and RT-PCR respectively (P < 0.05) in EGB treated group.</p><p><b>CONCLUSION</b>EGB may improve the learning and memory ability of diabetic rats the mechanism may be attributed to its improvement of the expression of NGF and NT-3 and reducing apoptosis in hippocampus neurons.</p>

Role of P38-MAPK signal transduction pathway and effect of panax notoginoside in rats with hypoxic hypercapnia pulmonary hypertension / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the role and significance of P38-MAPK in the pathological process of hypoxic hypercapnia pulmonary hypertension in rats, and the protection of panax notoginoside (PNS).</p><p><b>METHODS</b>(1) To set up rat pathological model of hypoxic hypercapnia pulmonary hypertension: seventy two male SD rats (200 280 g) were randomly divided into six groups (n = 12), which were normal group (N group), hypoxic hypercapnia for 3-day group (H3d), hypoxic hypercapnia for 1-week group(H1w), hypoxic hypercapnia for 2-week group (H2w), hypoxic hypercapnia for 4-week group (H4w) and PNS-injected group (Hp). The rats of PNS -injected group were injected PNS before being placed in the chamber (50 mg/(kg x d), ip), and other groups were injected normal sodium (2 ml/kg, ip). (2) The shapes of pulmonary artery were detected by HE staining. (3) Western blot was used to study the protein expression of p38-MAPK. The expression of p38-MAPK in lung tissue and pulmonary blood vessel was investigated by immunohistochemistry.</p><p><b>RESULTS</b>(1) The ratio of vessel wall area/total area (WA/ TA) in H1w, H2w, H4w and Hp group was higher than that of N group (P < 0.05), but that of H3d group did not change obviously (P > 0. 05 vs N group). The ratio of WA/TA in Hp group was obviously lower than that of H4w, group (P < 0.05). (2) The levels of P-p38 protein was markedly ascended in H3d group (0.225 +/- 0.071) compared with N group (0.012 +/- 0.006), and expression of P-p38 protein was significantly positive in H1w, H2w, H4w groups. (P < 0.05). (3) As P-p38 protein in pulmonary arterial tunica intima and tunica media, sterile expression in N group (0.099 +/- 0.015) and H3d group (0.107 +/- 0.013) contrasted to H4w group (0.124 +/- 0.025, P < 0.05), then tended to rise in H2w, H4w group (P < 0.05). (4) In pulmonary tissue, the levels of P-p38 protein in PNS-injected group were lower 53.02% (P < 0.05) than those in H4w group. In pulmonary arterial tunica intima and tunica media the levels of P-p38 protein in PNS-injected group were lower 87.33% (P < 0.05) than those in H4w group.</p><p><b>CONCLUSION</b>p38-MAPK as a signal transduction may play an important role in the development of hypoxia induced pulmonary hypertension. The effect of PNS on reducing pulmonary hypertension and improving pulmonary vascular wall remodeling may be related to its inhibiting expression of p38 MAPK.</p>

The role of JNK in apoptosis of renal tubular epithelial cells in diabetic rats with fluctuant high blood glucose / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the signal transduction mechanisms of apoptosis in renal tubular epithelial cells in diabetic rats with fluctuant high blood glucose.</p><p><b>METHODS</b>Healthy SD rats were randomly divided into 3 groups: normal control group (A), stable high blood glucose group (B) and fluctuant high blood glucose group (C). Diabetic rats were induced by intraperitoneal injection of streptozotocin (STZ, 65 mg/kg), and the fluctuant high blood glucose animal model was induced by intraperitoneal injection of ordinary insulin and glucose at different time point every day. The superoxide dismutase (SOD) activity and the content of malonaldehyde (MDA) in renal tissue homogenate were detected with colorimetry. The protein expression of Nox4 and JNK were examined by immunohistochemistry and Western blot. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL).</p><p><b>RESULTS</b>After 12 experimental weeks, significantly increased cell apoptosis, up-regulation of Nox4 and P-JNK expression in renal tubular epithelial cells were observed in B and C groups compared with those in A group. The MDA content increased and SOD activity decreased in renal tissue in B and C groups. Above effects were more obviously shown in C group.</p><p><b>CONCLUSION</b>Fluctuant high blood glucose induced more apoptosis of renal tubular epithelial cell than stable high blood glucose in diabetic kidney, which might be related to the activation of JNK signal transduction pathway.</p>

Association between polymorphisms of inflammatory factor genes and coronary heart disease / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association between genetic polymorphisms of inflammatory factors and susceptibility to coronary heart disease(CHD) in southern Chinese Han population.</p><p><b>METHODS</b>Using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) method, the genotypes of five inflammatory factors (BRCA1-associated protein, a disintegrin and metalloproteinase 8, inter-alpha-trypsin inhibitor H3, interleukin-15, cyclooxygenase-2) were anaylzed in 283 CHD patients diagnosed by angiography and 176 controls.</p><p><b>RESULTS</b>In these inflammatory factors, the 270T/C and 90A/G polymorphisms of the BRAP gene showed a significant association with CHD. The allele and genotype frequencies of BRAP gene were consistent with those predicted by Hardy-Weinberg equilibrium (chi-square=0.878, P> 0.05; chi-square=0.776, P> 0.05, respectively). The frequencecies of 270C and 90G alleles in CHD patients was significantly higher than those of the control group (29.51% vs. 21.31%, P=0.006; 30.04% vs. 21.31%, P=0.004, respectively). Compared with 270TT and 90AA, 270CC and 90GG genotypes had a significantly increased CHD risk by Logistic regression analysis (OR=4.51, 95%CI: 1.41-14.45, P=0.011; OR=5.09, 95%CI: 1.60-16.26, P=0.006, respectively). This association was still signifcant after adjustment for the sex, age, smoke, hypertension, diabetes, plasma total cholesterol and low density lipoprotein levels. No evidence of association was found for other single nucleotide polymorphisms.</p><p><b>CONCLUSION</b>The 270T/C and 90A/G polymorphisms in the BRAP gene may contribute to an increased risk of CHD among southern Chinese Han population.</p>

Effects of astragalus and angelica on bone marrow stem cells proliferation and VEGF protein expression in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To observe the effects of Astragalus and Angelica on bone marrow stem cells (BMSU) proliteratlon mn vitro and investigate its possible mechanism.</p><p><b>METHODS</b>Five 200 to 220 g SD rats were fed with a high fat diet for 4 weeks and given 30 mg/kg streptozotocin (STZ) twice develop type II diabetes from July 2009 to February 2010. The rats with blood glucose concentrations of 16.7 mmol/L or more were considered diabetic. Bone Marrow Stem Cells (BMSC) were collected and isolated by density gradient centrifugation. The BMSC were divided into 4 groups,including empty control group, Astragalus group, Angelica group and Astragalus plus Angelica group. DMEM of 100 microl was added in empty control group. DMEM of 100 microl containing Astragalus (1100 mg/L), Angelica (1100 mg/L) and Astragalus (1100 mg/L) combine with Angelica(220 mg/L) were added in Astragalus group, Angelica group and Astragalus plus Angelica group respectively. The cell proliferation was detected by MTT method, and the concentration of VEGF in the supernatant was determined by ELISA. The VEGF expression was analyzed by Western Blot after 14 days incubation.</p><p><b>RESULTS</b>The BMSC proliferation and the VEGF concentration in the supernatant and the BMSC VEGF protein expression significantly increased in Astragalus group and Astragalus plus Angelica group compared to those of empty control group (P < 0.05 or P < 0.01). The above effects were more strong in Astragalus plus Angelica group (P < 0.05).</p><p><b>CONCLUSION</b>Astragalus with Angelica or used separately could promote BMSC proliferation. The mechanism might induce the VEGF protein expression in BMSC. And the independent use of Angelica has no above effect.</p>

Effects of ambroxol combined with low-dose heparin on TNF-alpha and IL-1beta in rabbits with acute lung injury / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the intervention and mechanism of ambroxol combined with low-dose heparin on oxidative stress, TNF-alpha and IL-1beta in rabbits with acute lung injury (ALI).</p><p><b>METHODS</b>Twenty-four healthy Japanese rabbits were randomly divided into three groups: (1) Normal saline control group (NC), (2) Oleic acid injury group (OA), (3) Ambroxol + low-dose heparin therapy group (AH). After the success of ALI model, AH group was injected ambroxol + low-dose heparin, while the NC group and OA group were injected the same dose of normal saline by the same method. Arterial oxygen tension (PaO2), tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) at different time points were determined. The pathological manifestation of both side lungs was observed at the end of expeiment. The activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), xanthine oxidase (XO) and the content of malondialdehyde (MDA) in bronchoalveolar lavage fluid (BALF) and lung tissue homogenate were tested. The apoptosis index was detected. The lung wet/dry (W/D) ratio was calculated. The pathological changes in lung tissue were observed by light microscopy, and the ultrastructural changes of lung tissue were observed by electron microscopy.</p><p><b>RESULTS</b>(1) The instructive injury induced by ALI observed under electron microscope and light microscope and W/D was decreased significantly in AH group. (2) PaO2 was improved significantly in AH group, compared with that in OA group (P < 0.01). (3) The activity of GSH-Px and SOD in AH group increased significantly (P < 0.01 or P < 0.05) but the activity of XO and the content of MDA decreased significantly (P < 0.01), compared with those in OA group. (4) Except the content of IL-1beta in serum before treatment, the content of IL-1beta and TNF-alpha in serum, BALF, lung tissue homogenate of OA group increased significantly (P < 0.01), and those were obviously improved in AH group. (5) Apoptosis index (AI) in AH group decreased significantly (P < 0.01) compared with that in OA group.</p><p><b>CONCLUSION</b>In ALI induced by OA, IL-1beta and TNF-alpha increases significantly and involved in the occurrence and development of ALI. Ambroxol combined with low-dose heparin can reduce lung cells oxidative stress to inhibit the release of IL-1beta and TNF-alpha, which play a role in the treatment of ALI.</p>

Effects of benazepril on apoptosis and expression of Bax and Bcl-2 in kidney of diabetic rats with different blood glucose levels / 中华内分泌代谢杂志

Diabetic rats were induced by intraperitoneal injection of streptozotocin.TUNEL and immunohistochemistry results showed that the renal tubular cell apoptosis index(AI)and Bax protein expression were significantly reduced,and the Bcl-2 protein expression in glomeruli was significantly increased in diabetic rats with stable hyperglycemia treated by benazepril compared with diabetic rats with stable hyperglycemia treated by vehicle(all P