Effect of Qushi-Dizhuo decoction on urate transporters in hyperuricemia rats

AIM:To investigate the effects of Qushi-Dizhuo decoction (QSDZ) on the expression of uric acid transporters in rats with hyperuricemia. METHODS:Male Sprague-Dawley rats (n=60) were randomly divided into blank control group, model group, allopurinol group, and low-, middle- and high-dose QSDZ groups, with 10 rats in each. Except the blank control group, rats in the other groups were treated with adenine and potassium oxonate to prepare hyperuricemia rat model by intragastric administration once a day. Meanwhile, the rats in treatment groups were intragastric admi-nistrated with the corresponding preparation, after 28 days of continuous administration, the rats were sacrificed. Serum uric acid (UA) and creatinine (Cr) levels and xanthine oxidase (XOD) activity were measured. The morphological changes of renal tissues were observed by HE staining. The protein and mRNA expression of urate transporter1 (URAT1), glucose transporter 9 (GLUT9) and organic anion transporter 1 (OAT1) were detected by Western blot and real-time PCR. ATP-binding cassette subfamily Gmember 2 (ABCG2) and organic cation transporter 1 (OCT1) were also determined. RESULTS:Compared with the blank control group, the content of UA and Cr and the activity of XOD were significantly increased (P<0.05), the URAT1 and GLUT9 protein levels were significantly up-regulated, and the OAT1, ABCG2 and OCT1 protein levels were significantly down-regulated in model group (P<0.05). Compared with model group, the levels of serum UA and Cr and actioity of DXD were significantly decreased in QSDZ middle,high-dose group (P<0.05). The protein levels of URAT1 and GLUT9 were decreased, while the protein levels of OAT1, ABCG2 and OCT1 were increased in middle- and high-dose groups compared with model group (P<0.05). The mRNA expression of URAT1, GLUT9, OAT1, ABCG2, OCT1 was consistent with its protein expression trend. CONCLUSION:QSDZ decoction may through inhibiting the activity of XOD and regulating the expression of transporter to show its anti-hyperuricemia effect.     

Characteristics and pathomechanisms of hyperuricemia combined with hypertriglyceridemia and hyperglycemia in a coturnix model induced by high-purine diet

AIM: To investigate the pathomechanisms in a coturnix model of high-purine diet and the metabolic characteristics of glucose and lipids. METHODS: Twenty-four French male quails were randomly divided into 2 groups: control group and model group. The animals in control group were fed with normal diet and the quails in model group were fed with high-purine diet. The body weight, serum uric acid (UA), triglyceride (TG), glucose (GLU), the activity of xanthine oxidase (XOD), guanine deaminase (GuDa) and glyceraldehyde phosphate dehydrogenase (GAPDH), and the level of insulin (Ins) were determined. RESULTS: No change of body weight in model group was observed. In model group, the serum levels of UA,TG and GLU were significantly increased from 10 d to 140 d, 60 d to 140 d and 90 d to 140 d, respectively. At 10 d, 60 d and 140 d, the activity of XOD in model group was significantly higher than that in control group. From 30 d to 140 d, the activity of GAPDH was significantly decreased. From 60 d to 140 d, the level of Ins was significantly increased. CONCLUSION: (1) High-purine diet induces multiple metabolic disorders of UA, TG and GLU. (2) The pathologic processes can be divided into three stages: simple hyperuricemia in the first stage, hyperuricemia combined with hypertriglyceridemia in the second stage and hyperuricemia combined with hypertriglyceridemia and hyperglycemia in the third stage. (3) The pathomechanisms may relate to the increased activity of XOD, decreased activity of GAPDH and increased level of Ins.     

Protective effect of C1q/tumor necrosis factor-related protein 3 on vascular endothelium in rats with hyperuricemia

AIM To study whether C1q/tumor necrosis factor （TNF）-related protein 3 （CTRP3）protect vascular endothelium in rats with hyperuricemia and its potential mechanisms. METHODS An animal model of hyperuricemia was established by using male SD rats drinking 10% fructose water （n=10）. The rats drinking normal water served as normal controls （n=10）. After 12 weeks, the rats were given a single injection with Ad-CTRP3 or Ad-GFP. The experiment was ended at 14th day after transfection.The serum levels of uric acid and nitric oxide （NO） were evaluated. The serum contents of TNF-α and interleukin-6 （IL-6） were measured by ELISA. HE staining and TUNEL assay were used to assess the morphological changes of intima and apoptosis of endothelial cells in thoracic aorta, respectively. The mRNA levels of endothelial nitric oxide synthase （eNOS）, TNF-α and IL-6 were detected by RT-qPCR. The protein levels of CTRP3 and Toll-like receptor 4 （TLR4） were determined by Western blot. RESULTS Compared with normal control group, the rats with hyperuricemia showed lower CTRP3 and higher TLR4 protein levels in the thoracic aorta （P<0.05）. Hyperuricemic rats had higher serum contents of uric acid, TNF-α and IL-6 （P<0.05）. Also, the intima structure disturbance of thoracic aorta, increased apoptotic rate, higher mRNA levels of TNF-α and IL-6 as well as lower mRNA levels of eNOS were observed （P<0.05）. By contrast, CTRP3 over-expression decreased TLR4 protein levels, reduced inflammatory cytokines, and obviously improved the morphology and function of thoracic aorta in the rats with hyperuricemia. CONCLUSION CTRP3 protect vascular endothelium in rats with hyperuricemia maybe via down-regulation of TLR4- mediated inflammatory signaling pathway.     

Role of kisspeptin/GPR54 signaling pathways in prompting formation of precocious puberty

AIM: To explore the role of kisspeptin/GPR54 signaling pathway in the pathogenesis of precocious puberty based on female precocious puberty rat model induced by the single dose of danazol. METHODS: Female SD rats aged 3 d were randomly divided into normal group, vehicle group and model group. On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture. All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development. The levels of E₂, FSH and LH in peripheral blood were measured by ELISA. The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR. The kisspeptin expression in rat hypothalamus was observed by immunofluorescence. RESULTS: The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group. On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group. On days 25 and 30, the ovarian morphological development in the model rats was significantly earlier than that in normal group. On day 25, the mRNA expression of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats significantly increased compared with normal group and vehicle group. On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group. On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group. The mRNA expression of GPR54 had no obvious difference among all groups. CONCLUSION: The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty, suggesting that kisspeptin is an initiating factor for precocious puberty. Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty.     

Effect of oleanolic acid on expression of TNF-α and collagen in silicotic rats in vivo

AIM: To observe the effect of oleanolic acid (OA) on the expression of Tumor necrosis factor-α (TNF-α) and collagen in silicotic rats in vivo and its possible mechanism. METHODS: Male Wistar rats were divided into 4 groups according to the randomized block design: control group, model group, OA group and solvent control group (20 rats in each group). Except control group, the rats in other groups were induced by intratracheal instillation of silicon di-oxide (SiO₂; 250 mg/kg). The rats in OA group were intragastrically administered with OA (60 mg/kg) from the second day of giving SiO₂. The rats in solvent control group were gavaged daily with 0.6% sodium carboxymethyl cellulose solution (10 mL/kg). The rats in control group were given normal saline under the same condition for 56 consecutive days. All rats were killed at the 7th, 14th, 28th and 56th days. The lung coefficient was detected and the morphological changes were observed. The serum contents of TNF-α were detected by ELISA. The content of total collagen in the lung tissue was measured. The protein level of nuclear factor-κB (NF-κB) in the lung tissue was determined by immunohistochemical method. RESULTS: (1) According to the morphological changes, the silicosis model was successfully established. Compared with control group, the lung coefficient and total collagen increased obviously in model group and solvent control group. The lung coefficient and total collagen content in OA group at each time point reduced compared with those in model group and solvent group, and increased compared with those in control group at the corresponding time points. (2) The serum contents of TNF-α in model group and solvent control group significantly increased, peaking at the 14th day, slightly decreasing afterward, and showing statistically significant difference at each time point compared with those in control group. No significant difference between model group and solvent group at different time points was observed. OA had inhibitory effect on the contents of TNF-α compared with model group and solvent group at the corresponding time points. (3) NF-κB in model group and solvent control group significantly increased, peaking at the 28th day, and showing statistically significant difference at each time point compared with those in control group. The NF-κB expression in OA group was similar to model group, but significantly decreased compared with control group at each time point. CONCLUSION: OA inhibits the expression of TNF-α and collagen and attenuates the silicosis fibrosis, which may be related to the NF-κB pathway.     

Imbalance between matrix metalloproteinases and tissue inhibitor of metalloproteinases during wound healing in diabetic rats

AIM: To investigate the imbalance between the expression of metalloproteinases (MMPs) and that of tissue inhibitors of metalloproteinase (TIMPs) during wound healing in diabetic rats. METHODS: Diabetic rats were induced with streptozotocin. All rats were maintained for 6 weeks. A full-thickness excisional wound was created on the back of each rat. Every group was randomly divided into 3 subgroups of 7 rats: 3 d group, 7 d group, 14 d group and animals were killed at 3rd, 7th and 14th day. Routine pathological examination, Masson′s trichrome staining and immunohistochemistry were made to calculate the score of epidermal and dermal regeneration, granulation tissue thickness, angiogenesis, matrix density, and infiltrated cells at different time points. RT-PCR and Western blotting were used to detect the expression of mRNA and protein of MMP-9 and TIMP-1 in the skin at those time points. RESULTS: Six weeks after streptozotocin treatment, Three days after injury, the wound healing rate of normal rats was faster than that of diabetic rats. From 3rd to 14th day, there were a lot of fibroblast and macrophage in normal skin, while few such cells were observed in diabetic skin. The other histological scores in normal skin were higher than those in diabetic rats at 7th and 14th day. Both MMP-9 and TIMP-1 had minimally detectable levels before wounding but exhibited rapid, significantly large increases within 3 d after wounding. Subsequently, they showed a rapid decline by 14 d. The relative values of expression of MMP-9 mRNA and protein in diabetic group were higher than those in normal group at different time points. However, the values of TIMP-1 mRNA and protein in diabetic group were significantly lower than those in control group. Significant difference was observed between two groups with the ratio of MMP-9/TIMP-1, higher in diabetic group than that in normal group. CONCLUSION: Abnormal reepithelialization, angiogenesis, inflammatory cell infiltration, collagen fibers generation, granulation tissue deposition, seem to be the basic histopathology that delays wound healing. The imbalance between MMPs and TIMPs in diabetic skin tissue before and after injury may be one of the important reasons of these alterations of histopathology.     

Protective effect of activated macrophages on retinal ganglion cells after optic nerve crush

AIM:To investigate effects of factors derived from activated macrophages on survival of retinal ganglion cells (RGCs) after optic nerve crush. METHODS:Adult Wistar rats were randomly divided into normal control, crush control, medium treatment and zymosan treatment groups. All the rats were retrogradly by injecting fluorogold into superior colliculi of both sides. Partial optic nerve injury model was induced in the left eyes by a special designed optic nerve clip with 40 gram power for 9 seconds at optic nerve 2 mm behind the eyeball except for normal control group. The numbers of the labeled RGCs in each group were counted. RESULTS:The numbers of RGCs in crush group was 92.6%, 82.9%, 72.6% and 57.6% of normal controls on the third, 7th, 15th and 21th day, respectively. In the group with zymosan injection, the number of RGCs were much higher than that in controls on the third, 7th, 15th and 21th day (P<0.01). CONCLUSION:Intravitreal injection of zymosan activates macrophages. Macrophages-derived factors might protect RGCs after optic nerve crush and improve their survival.     

Effect of hyperbaric oxygen on HIF-1α expression in rat experimental periodontitis with psychological stress

AIM: To investigate the effect of hyperbaric oxygen (HBO) on hypoxia-inducible factor-1α (HIF-1α) expression in rat experimental periodontitis with psychological stress. METHODS: Male special pathogen-free Wistar rats (n=120) were randomly divided into 4 groups: normal control group; psychological stress stimulation group; experimental periodontitis group: the periodontitis model was induced by wrapping 3/0 silk ligature inoculated with Porphyromonas gingivalis around the left maxillary second molar of the rats; periodontitis model with stress stimulation group. Psychological stress was removed at the 9th weeks after ligature, 6 rats from each experiment group were randomly chosen to HBO treatment. The rats were sacrificed at the 2nd, 4th, 8th and 10th weeks after ligature. Gingival index (GI) and attachment loss (AL) were measured before sacrifice. The histological changes of periodontal tissues were observed under microscope with HE staining. The expression of HIF-1α was observed by the method of immunohistochemistry. RESULTS: The sites of gingival attachment were normal in control group and psychological stress stimulation group. Periodontal pocket, and periodontal attachment loss were observed in experimental periodontitis group. The tissue damage was much serious in periodontitis model with stress stimulation group. No significant difference of GI and AL among psychological stress stimulation group and normal control group during the experiment was observed. GI and AL in periodonitis model with stress stimulating group were significantly higher than those in experimental periodontitis group at the 4th and 8th weeks (P < 0.01). The levels of GI and AL were significantly lower at the 10th weeks after HBO treatmnt than those in untreated groups (P < 0.05). No significant difference of HIF-1α expression scores among psychological stress stimulation group and normal control group was found. HIF-1α expression scores in periodonitis model with stress stimulating group was significantly higher than that in experimental periodontitis group at the 4th and 8th weeks (P < 0.01). At the 10th weeks after HBO treatment the levels of HIF-1α were significantly lower than that in untreated groups (P < 0.01). CONCLUSION: Stress stimulation may aggravate periodontitis by decreasing tissue oxygenation in rats. HBO may represent a useful way in psychological stress periodontitis therapy.     

Expression of urotensin II and its receptor in nephridial tissues of rats with acute renal damage

AIM: To observe the mRNA expression of urotensin II (UII) and its receptor (G protein-coupled receptor 14,GPR14) in nephridial tissues of rats with acute renal damage. METHODS: Male Wistar rats were divided into 2 groups: the rats in control group (n=10) were administered with normal saline by gavage; the rats in model group (n=30) were administered with Caulis Aristolochiae manshuriensis (CAM) by gavage for 25 d to induce acute renal damage. Every 5 rats in model group were sacrificed on the 3rd, 7th, 15th and 25th days during CAM treatment and all rats in the 2 groups were killed 10 d after withdraw of CAM. The kidneys were collected for pathological observation and the UII and GPR14 mRNA examination.RESULTS: The degeneration, necrosis and disintegration in tubules were observed as major pathological changes in the rat kidneys after 3 d of CAM administration. The pathological changes were aggravated following the duration of in CAM administration, and were remained and even worsen when CAM was withdrawn for 10 d. Compared with control group, the mRNA expression of UII was significantly elevated (P<0.05) at the time point of CAM administration for 15 d,even obviously increased (P<0.01) at the time point of CAM administration for 25 d, and remained at the highest levels to the end of the observation. The mRNA expression of GPR14 was significantly increased (P<0.05) at the time point of CAM administration for 7 d, became higher (P<0.01) on the 15th day, and gradually increased as the experimental time went on. CONCLUSION: The mRNA levels of UII and its receptor are significantly elevated in CAM-induced renal lesion in rats, suggesting that UII plays a pathological role in the development of acute renal damage.     

Mechanisms of hyperglycemia induced by immunosuppressant FK506

AIM：To investigate the effect of immunosuppressant FK506 on serum glucose in rats and to explore its mechanism. METHODS：Sprague-Dawley rats (n=12) were randomly divided into drug group and normal group. The rats in drug group were intraperitoneally injected with FK506 at dose of 1 mg·kg-1·d-1 and the rats in normal group received saline (1 mL·kg-1·d-1, ip) for 14 d. The fasting weight and fasting glucose were regularly measured every 2 d. Visceral fat was isolated from the rats at the end of experiment. The mRNA expression of adiponectin, leptin, visfatin, resistin, retinol-binding protein 4 (RBP4) and peroxisome proliferator-activated receptors γ (PPAR-γ) was determined by real-time fluorescence quantitative PCR. The protein expression of PPAR-γ and adiponectin was measured by Western blotting. RESULTS：Compared with normal group, the concentration of fasting blood glucose in model group was significantly increased from the 10th day (P<0.05). At day 14, the fasting blood glucose of the model group increased from (5.10±062) mmol/L to (7.73 ± 0.73) mmol/L. No significant change of blood glucose in normal group between the 10th day and the 14th day ［from (4.66 ± 0.32) mmol/L to (5.80±0.10) mmol/L］ was observed. Compared with normal group, the mRNA expression of PPAR-γ, adiponectin and leptin in the adipose tissue of model group was significantly decreased (P<001), whereas the expression of visfatin, resistin and RBP4 was significantly increased (P<005). Compared with normal group, the expression of PPAR-γ and adiponectin in model group was decreased (P<001). CONCLUSION：FK506 may decrease the expression of PPAR-γ to change the expression of adipocytokines and induce hyperglycemia in rats.     

Effects of tripterygium glycosides combined with dexamethasone on TLRs/NF-κB signaling pathway in EAE rats

AIM: To investigate the role of TLRs/NF-κB pathway in experimental allergic encephalomyelitis (EAE) rats treated with tripterygium glycosides (TG) + dexamethasone (DX). METHODS: Lewis rats were used in the study and divided into control group, EAE model group, therapy 1 group (EAE rats treated with DX) and therapy 2 group (EAE rats treated with DX+TG). The mean clinical score of the rats was determined. The expression of TLR4 and TLR9 at mRNA and protein levels was detected by the methods of real-time quantitative RT-PCR and immunohistochemistry. The protein level of NF-κB p65 was also measured. The levels of TNF-α, IL-1β and IL-6 were assayed by ELISA. RESULTS: The mean clinical scores at 5th, 16th and 20th day were lower in therapy 1 group and therapy 2 group than that in EAE model group. The mean clinical score in therapy 2 group was even lower than that in therapy 1 group. At the 16th day (the peaking period), the mRNA expression of TLR4 and TLR9 in therapy 1 group and therapy 2 group were obviously lower than that in EAE model group. The protein levels of TLR4, TLR9 and NF-κB p65 were also significantly lower in therapy 1 group and therapy 2 group than those in EAE model group at peak stage of EAE. The levels of TNF-α, IL-1β and IL-6 were lower in therapy1 group and therapy2 group than those in EAE model group. The significant differences of the mean clinical score, the mRNA expression of TLR4 and TLR9, the positive ratio of NF-κB p65 and the levels of TNF-α, IL-1β and IL-6 between therapy 1 group and therapy 2 group were found. The result of orthogonal factorial analysis of variance indicated that the difference of therapeutic effect between DX and DX+TG was significant (F=75.749, P<0.01). CONCLUSION: The TLRs/NF-κB pathway takes part in the pathological process of EAE. TG combined with DX alleviates the symptoms of EAE by suppressing inflammatory and immunological reactions of EAE.     

VPA alleviates bleomycin-induced pulmonary fibrosis in rats

AIM: To investigate the effect of sodium valproate (VPA) on bleomycin-induced pulmonary fibrosis (PF) and its mechanism. METHODS: SD rats (n=42) were randomly divided into model group and treatment group. Bleomycin at dose of 5 mg/kg was intratracheally injected to establish a rat PF model. The rats in treatment group were given normal saline (NS, 0.5 mL/d), VPA (300 mg·kg^-1·d^-1) or dexamethasone (DEX, 0.6 mg·kg^-1·d^-1) via intraperitoneal injection from the 14th day to the 28th day after modeling. The rats in model group were sacrificed on 0 d, 14 day and 28 d after modeling . The rats in treatment group were killed at 14th day after treatment. The effects of VPA on PF were evaluated by HE and Masson staining. The hydroxyproline (HYP) content in the rat lung tissues was detected, and the expression of α-smooth muscle actin (α-SMA) and E-cadherin was determined by Western blotting. RESULTS: HE staining showed that the alveolar structure and interstitial morphology in VPA group were better than those in NS group and DEX group. The level of collagen in VPA group was significantly lower than that in DEX group and NS group by determining the HYP content and Masson staining. VPA reduced the expression of α-SMA, a mesenchymal marker protein of PF, while increased the expression of epithelial marker protein E-cadherin. CONCLUSION: VPA reduces collagen content and distribution, and up-regulates the expression of the epithelial marker protein E-cadherin, while down-regulates mesenchymal marker protein α-SMA, thereby preventing the rat lung tissues from bleomycin-induced PF.     

Influence of leptin on intestinal function and its protective effect on hepa tic and renal functions after sepsis

AIM: To detect the effect of sepsis on hepatic,renal functions and corresponding enzymes in intestine of mice,and to explore the role of leptin in acute inflammation.METHODS: A mice model of sepsis was made by cecum ligation and perforation,and 96-well spectrophotometry was used to detecte the levels of alanine aminotransferase (ALT),uric acid (UA) and activities of myeloperoxidase (MPO),glutathin-S-transferase (GST),xanthine oxidase (XOD),superoxide dismutase (SOD) in serum and intestinal homogenized fluids,respectively.Hematoxylin-eosin staining was used simultaneously to check the histopathologic changes of intestine.RESULTS: Compared with sham group (330.12 μmol/L±94.15 μmol/L),serum UA level (521.92 μmol/L±91.86 μmol/L) at 6 h after sepsis was significantly higher.12 h after sepsis,both serum ALT (83.55 U/L±40.44 U/L) and UA (474.03 μmol/L±75.22 μmol/L) were significantly higher than those in sham group (66.23 U/L±16.80 U/L and 320.95 μmol/L±99.14 μmol/L,respectively).12 h after leptin injection (0.1 mg/kg,ip) or indomethacin injection (2 mg/kg,ip),the serum ALT and UA levels significantly decreased (vs sepsis group,P<0.05).Moreover,the activities of MPO,GST,XOD and SOD in intestine were changed at different degrees.CONCLUSION: During the process of sepsis,trace dose of leptin injected peritoneally significantly improves and stabilizes the hepatic and renal functions.The mechanisms may be related with those intestinal enzymes associated with metabolism of free radicals,mercapto group and purine.Indomethacin exerts a similar role as leptin though at much higher dose.     

Effect of hyperbaric oxygen on periodontitis with psychological stress in rats

AIM: To investigate the role of chronic psychological stress on periodontitis and the effects of hyperbaric oxygen (HBO) on periodontitis with psychological stress in rats. METHODS: Male special pathogen-free Wistar rats (n=80) were randomly divided into 4 groups: (1)normal control group; (2)experimental periodontitis group: the periodontitis model was induced by wrapping 3/0 silk ligature inoculated with Porphyromonas gingivalis around the left maxillary second molar of the rats; (3)psychological stress stimulation group; (4)periodontitis model with stress stimulation group. Psychological stress was removed at the 9th week after ligature, and 4 rats from each experimental group were randomly chosen for HBO treatment. The rats were sacrificed at the 2nd, 4th, 8th and 10th weeks after ligature. The levels of blood glucose, adrenocorticotropic hormone (ACTH), corticosterone and adrenaline were measured as the stress markers. The histological changes of periodontal tissues were observed under microscope with HE staining. RESULTS: The levels of blood glucose, ACTH, corticosterone and adrenaline in psychological stress stimulation group and periodontitis with stress group were significantly higher than those in control group and experimental periodontitis group at the 2nd and 4th weeks after ligature (P<0.05). The levels of the stress markers were significantly lower than those in untreated groups in the 10th week after HBO (P<0.01). The sites of gingival attachment were normal in control group and psychological stress stimulation group. Periodontal pocket, and periodontal attachment loss (AL) were observed in experimental periodontitis group. The tissue damage was much heavier in periodontitis model with stress stimulation group as the furcation of tooth was exposed and the tissue damage was observed on both sides of the adjacent teeth. No significant difference of AL between psychological stress stimulation group and normal control group during the experiment was observed. The AL in periodontal model with stress stimulating group was significantly higher than that in experimental periodontitis group at the 2nd, 4th and 8th weeks (P<0.01). The level of AL was attenuated at the 10th week after HBO (P<0.01). No difference of histological change in periodontal tissues was observed between control group and psychological stress stimulation group. Severer inflammatory changes and alveolar bone destruction were observed in periodontitis with stress group than those in experimental periodontitis group. The levels of inflammation reduced at the 10th week after HBO. CONCLUSION: Stress stimulation is one of the inducing factors of periodontitis in rats, which aggravates periodontitis. HBO may represent a useful way in treating psychological stress periodontitis.     

Effect of hypothalamic arcuate nucleus damaged by monosodium glutamate on bone histomorphometry parameters in rat

AIM: To investigate the effect of hypothalamic arcuate nucleus(ARC) damaged by monosodium glutamate (MSG) on bone histomorphometry parameters in rats. METHODS: (1) Newborn SD rats of experimental group were hypodermically injected 10% MSG (4 g/kg BW) on the postnatal lst, 3rd, 5th, 7th, 9th day. Meanwhile, the newborn rats of the control group were given equal volume of normal saline and the rats after the postnatal 70th, 72th, 74th, 76th and 78th day were hypodermically injected 10% MSG (4 g/kg BW) as the MSG toxicity control group. After survivial for 160 days, all rats were killed. (2) Morphological methods were used to estimate the ARC neurons and the bone histomorphometry parameters. (3) Radioimmunoassay was used to measure the levels of the serum growth hormoen(GH), estradiol(E2) and testosterone(T). RESULTS: The number of the hypothalamic arcuate nucleus neurons, the levels of serum E2, T, GH and the %Tb·Ar, Tb·Th, Tb·N of proximal tibial metaphysis(PTM) significantly decreased, while Tb·Sp of proximal tibial metaphysis(PTM) significantly increased, and the osteoporotic alterations appeared obviously. All these changes did not appear in the rats of NS group and MSG toxicity control group. CONCLUSION: (1) The changes of the bone in MSG guoup rats are not the effect of the MSG toxicity on the bone directly. (2) The hypothalamic arcuate nucleus participates in the regulation of the bone metabolism. (3) ARC regulates bone metabolism via altering the functions of the hypothalamus-GH-IGF-1 axis and the hypothalamus-pituitary-gonal axis.     

Effect of Yangyu Tuji on content of type Ⅰ, Ⅲ collagen and the expression of MMPs and TIMP-1 in wound caused by streptozotocin in rats

AIM: To study the effects of Yangyu Tuji (YYTJ) on delayed healing wound of diabetic rats caused by streptozotocin (STZ). METHODS: SD male rats were randomly divided into control group (control), model group (model); and 3 different dose groups of YYTJ. 55 mg/kg STZ were given by intraperitoneal injection except for control group. After 30 days, a round skin of 1.6 cm diametre was excised on all dorsal back of rats. The healing time and healing rate were observed according to re-epithelization. The content of collagenⅠ and Ⅲ was observed by Picric acid-Sirius red staining , Matrix metalloproteinase-1, 13 (MMP-1, -13), tissue inhibitor of metalloproteinases-1 (TIMP-1) by immuno-histochemistry assay. All data were analyzed by IPP software. RESULTS: The healing time in each group treated with YYTJ was shorter than that in model group (P<0.01), and the healing rate was increased (P<0.01, P<0.05). Content of type I collagen, ratio of type Ⅰ and Ⅲ collagen of high and mid dose group were significantly higher than that in model group (P<0.01) at 3rd, 7th, 11th day. The expression of MMP-1, -13 of each groups were higher than that in model group at 7th day (P<0.01, P<0.05), and MMP-1 trend to equal with model group at 11th day. MMP-13 was significantly lower than that in model group at 11th day (P<0.01, P<0.05). TIMP-1 of each group of wound was higher than that in model group at 3rd, 7th, 11th day (P<0.01, P<0.05). The ratio of type Ⅰ and Ⅲ collagens in each group was lower than that in model group at 11th day (P<0.01). Ratio of MMP-13 and TIMP-1 of high dose group and mid dose group were higher than that in model group at 3rd and 7th day (P<0.01). The ratio of each group was lower than that in model group at 11th day (P<0.01). Meanwhile, ratio of MMP-13 and TIMP-1 of high dose group and mid dose group were lower than that of lower dose group (P<0.05). CONCLUSION: It is possible that YYTJ accelerates wound healing by increasing collagen content of type Ⅰ and Ⅲ, especially type Ⅰ, as well as improves collagen deposition by regulating the balance of MMP and TIMP.     

Influence of Tangshen formula on endothelial function and hemorheology in diabetic nephropathy rats

ATM: To explore the influence of Tangshen formula (TS) on endothelial function and blood rheology in diabetic nephropathy (DN) rats. METHODS: The DN rat model was established by intravenous injection of low-dose (30 mg/kg) streptozotocin (STZ) after having the high-fat/high-glucose diets for one month. The animals were divi-ded into DN model group, TS group and valsartan group. Fasting blood glucose (FBG), serum total cholesterol (TC), serum triglyceride (TG), renal cortex blood flow and hemorheology were monitored. The content of von Willebrand's factor (vWF) and plasminogen activator inhibitor-1 (PAI-1) in the serum was determined by ELISA. RESULTS: Compared with normal group, FBG,TC,TG, vWF and PAI-1 were increased in DN model group (P<0.05), and no significant difference of FBG was observed. Compared with normal group, plasma viscosity, Casson viscosity, whole blood high/medium/low-shear viscosity, erythrocyte aggregation index, erythrocyte rigidity index and erythrocyte electrophoresis time were increased, and erythrocyte deformation index was decreased in DN model group (P<0.01). Compared with DN model group, plasma viscosity, Casson viscosity, whole blood high/medium/low-shear viscosity, erythrocyte aggregation index, erythrocyte rigidity index and erythrocyte electrophoresis time were decreased (P<0.05), but there was no significant difference for erythrocyte deformation index in TS group. Compared with normal group, the renal cortex microcirculation blood flow in DN model group was significantly decreased. Compared with DN model group, the renal cortex microcirculation blood flow was significantly increased in TS group (P<0.05), and no significant change in valsartan group was found.CONCLUSION: Tangshen formula plays a protective role in the kidney of diabetic rats by improving the blood rheology and endothelial function, thus ameliorating the renal cortex microcirculation blood flow in experimental diabetic rats.     

Effect of zhiganning on the expressions of HSP60 and HSP70 in the hepatocytes of rats undergoing steatohepatitis

AIM: To observe the expressions of HPS60 and HPS70 in hepatocytes in rats under treatment with zhiganning on steatohepatitis. METHODS: Male SD rats were randomly divided into large dose zhiganning group, small dose zhiganning group, ursodeoxycholic acid (UDCA) group, model group, normal control group. Except the normal control group, all the other rats were fed with high fat (88% standard diet, 10% lard, 2% cholesterol) and 35% alcohol 10 mL/kg twice a day. Prophylactic drugs were used at the same time. All rats were sacrificed at the 9th week. Routine histologic features of hepatic sections were observed by HE staining and penetrated electron microscope. The expressions of HSP60 and HSP70 in the liver were detected by immunohistochemistry, respectively. RESULTS: ⑴ The degree of steatohepatitis in the large dose zhiganning group and ursodeoxycholic acid (UDCA) group were significantly decreased compared with that in model group (P<0.05). ⑵ The expression of HSP70 in the large dose zhiganning group and ursodeoxycholic acid (UDCA) group were significantly higher than that in either model group or normal control group (P<0.05, P<0.01, respectively). The expression of HSP60 in the large dose zhiganning group was significantly higher than that in either model group or normal control group (P<0.05, P<0.01, respectively). ⑶ In the large dose zhiganning group and ursodeoxycholic acid group, ultramicroscopic structure of liver was nearly normal, which was significantly improved compared with model group. CONCLUSION: The results indicate that zhiganning and UDCA effectively prevente the steatohepatitis in rats induced by high fat diet and alcohol. The enhanced expressions of HSP60 and HSP70 may play an important role in the prevention of liver from injury.     

The renal protective effects of angiotensinⅡ typeⅠreceptor antagonist and angiotensin-converting enzyme inhibitor and their influences on intrarenal renin-angiotensin system

AIM:To compare the renoprotective effects of angiotensinⅡtypeⅠreceptor antagonist (AT₁RA) with that of angiotensin converting enzyme inhibitor (ACEI) and to investigate their influences on intrarenal renin-angiotensin system. METHODS: Experimental nephrotic syndrome model was induced in SD rats with repeated peritoneal injections of puromycin. Twenty-eight rats were randomly divided into four groups: normal control, nephrotic control, ACEI-treated and AT₁RA-treated group. Serum, urine and renal tissue were collected for study 12 weeks later. RESULTS: The urine protein was less and renal function was better in both treated groups. The glomerular and interstitial injury indexes of both ACEI and AT₁RA treated rats were lower than that of nephrotic control rats and had no significant difference between the two treated groups. The renal local ACE activity and angiotensinⅡ of nephrotic control group were significantly higher than that of normal control group and the two treated group(P＜0.01). CONCLUSION:AT₁RA and ACEI have comparable renal protective effects on nephrotic syndrome and these effects may be related to the inhibition of renal local ANGⅡ.