Effects of paecilomyces cicadidae total polysaccharides on macrophages of old rats

AIM: To study the activation and the immune regulation of paecilomyces cicadidae total polysaccharides on macrophages in old rats. METHODS: The ability of devouring neutral red and stimulating index in the alveolar macrophages (AM) and peritoneal macrophages (PM) were observed. The activities of lactate dehydrogenase (LDH), acid phosphatase (ACP) and arginase (Arg) were detected by automatic biochemistry analyzer. The shape and structure of spleen macrophages were observed by electron microscope. RESULTS: Compared with control group, the activity of the LDH/ACP/Arg in the AM or PM and in the culture fluid of the AM or PM were significantly increased (P<0.05 or P<0.01), and the ability devouring neutral red and stimulating index in the PM were strengthened (P<0.01), the ergastoplasm was hyperplasia and the amount of lysosome was manifold in the spleen macrophages of old rats induced by paecilomyces cicadidae total polysaccharides. CONCLUSION: Paecilomyces cicadidae total polysaccharides promote immune function in old rats.     

Immunomodulation of paecilomyces cicadidae polysaccharides on immune function of rats

AIM: To explore the immunomodulatory effect of paecilomyces cicadidae polysaccharides (PCPS).METHODS: Subcutaneous injection with 50, l00, 200 mg/kg of PCPS were given in the back of the rats everyday for l5 days. The number of white blood cells (WBC) was counted. The activities of acid phosphatase (ACP) and lactase dehydrogenase (LDH)in liver, kidney, spleen and thymus were detected by automatic biochemistry analyzer. The ability of devouring neutral red and activity of ACP, LDH, arginase in alveolar macrophages were also detected. The body weight of the rat everyday during experiment and weight of the spleen and thymus after the rats were killed were measured and wet weight index was calculated.RESULTS: The wet weight index of spleen and thymus, the activity of ACP, LDH, arginase and ability of devouring neutral red in alveolar macrophages in the test group treated with PCPS were significantly higher than those in control group. CONCLUSION: PCPS shows a significant immunomodulatory effect with the increasing counts of WBC and activation of alveolar macrophages in a dose-dependent manner.     

Paecilomyces cicadidae total polysaccharides regulates immune function in immunosuppressed rats

AIM: To study the effects of paecilomyces cicadidae total polysaccharides on the immune function and free radical scavenging of tissues in immunosuppressed rats. METHODS: The activities of lactate dehydrogenase (LDH) and arginase (Arg) in liver， kidney， spleen， thymus of immunosuppressed rats were detected by automatic biochemistry analyzer. The content of lipid peroxide (LPO) and the reduced glutathione (GSH) level in liver， kidney， spleen， thymus of immunosuppressed rats were observed. RESULTS: The activity of the LDH, Arg and GSH level were significantly increased, and the LPO content were significantly declined in liver， kidney， spleen， thymus of immunosuppressed rats induced by cyclophosphamide. CONCLUSION: Paecilomyces cicadidae total polysaccharides promote immune function and the ability of free radical scavenging of tissues in immunosuppressed rats.     

Effects of achyranthes bidentata polysaccharides on non-specific immune function of old rats

AIM: To study the effect of achyranthes bidentata polysaccharide (ABPS) on non-specific immune function of old rats. METHODS: After subcutaneous injection of ABPS (50 mg/kg body weight) into the back of the old rats for 21 days, the WBC and the levels of hemoglobin(Hb) and chief biochemical index in blood of old rats were determined. After the alveolar macrophages (AMΦ) and peritoneal macrophages (PMΦ) cultured with 10% fetal bovine serum-RPMI-1640 for 2 h in vitro, the activities of lactate dehydrogenase(LDH) and acid phosphatase(ACP) in the AMΦ and PMΦ were detected by automatic biochemistry analyzer, and the abilities in devouring neutral red in the AMΦ and PMΦ were observed. The levels of lipid peroxide(LPO) and reduced glutathione(GSH) in some immune organs like spleen, liver, brain, kidney and in serum were also detected.RESULTS: The levels of ACP and LDH in the AMΦ or PMΦ were increased (P<0.01), the ability of AMΦ or PMΦ in devouring neutral red was increased (P<0.01), the levels of LPO were diminished and the contents of GSH raised in the liver, brain and spleen (P<0.05 or P<0.01). CONCLUSION: ABPS has the functions of activating AMΦ and PMΦ， and enhances non-specific immunity in old rats.     

The activative effect of Paecilomyces cicadaeon peritoneal and alveolar macrophages of rats

AIM:To explore the activative effect of water extract from cultured mycelium ofPaecilomyces cicadae (P. cicadae)on peritoneal macrophages (PMΦ) and alveolar macrophages (AMΦ) of rats.METHODS:The rats were randomly divided into four groups: normal control group(Ⅰ), cyclophosphamide (Cy) group (Ⅱ),P. cicadaegroup(Ⅲ), Cy+P.cicadaegroup(Ⅳ).The rats were bred in the same circumstance and were administered with corresponding drugs by subcutaneous injection for 26 days. PMΦ and AMΦ of rats were irrigated by normal saline and collected by centrifuge and incubated in a humidified incubator for 2 h at 37 ℃. The activities of acid phosphatase (ACP) and lactate dehydrogenase (LDH) in the PMΦ and AMΦ were determined by biochemical methods, and the capability of PMΦ and AMΦ in phagocytosis of neutral red were measured by colorimetric method too. RESULTS:After administering P. cicadaeto rats, the activities of ACP and LDH in PMΦ and AMΦ of normal rats were elevated significantly,and the reduction of the activities of ACP and LDH in PMΦ and AMΦ of rats due to Cy was notably antagonized, and the capability of PMΦ and AMΦ phagocytizing the neutral red were strengthened significantly.CONCLUSION:P. cicadaecan activate the PMΦ and AMΦ of rats.     

Experimental studies on immunomodulatory and antitumor activity of polysaccharide from Paecilomyces cicadidae

AIM: To study the immunomodulatory and antitumor activity of both total polysaccharide from Paecilomyces cicadidae (PcPSt) and its fraction PcPSAc. METHODS: Five groups of mice which were composed of normal control, tumor control, group treated by PcPSt, treated by cyclophosphamide (Cy) and treated by both PcPSt and Cy were administrated by injecting to abdominis cavitas with normal saline (NS), NS, PcPSt , Cy , PcPSt+Cy correspondingly and respectively for 10 days after they had been injected B16 cell line except normal control.The white blood cells (WBC) were counted, spleen index and weight of tumor were statistically analysed. The cell counting kit 8 (CCK-8) was used to analyze proliferative activity of the cultured splenocytes. Nitric oxide (NO) kit was used to detect NO content of supernatant in each microplate. RESULTS: PcPSt increased WBC and relieved the decrease of WBC caused by Cy in tumor-bearing mice. PcPSt increasd the spleen index of tumor-bearing mice and cooperated with Cy to promote antitumor activity. PcPSAc at concentrations of 600 mg/L and 300 mg/L enhanced the proliferative activity of cultured splenocytes. Appropriate doses (15-300 mg/L) of PcPSAc promoted the secretion of NO, the effect of 300 mg/L of PcPSAc was the strongest. CONCLUSION: Paecilomyces cicadidae polysaccharide can promote immunomodulatory and antitumor activity in vivo and in vitro experiments.     

The activation effect of maize pollen polysaccharides on human thoracic cavity macrophages

AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTMΦ). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTMΦ were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-α) and interleukin-6(IL-6) in the hTMΦ was analyzed by radioimmunoassay, after hTMΦ were cultured with 0.312, 0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTMΦ induced by PPM, and the levels of TNF-α and IL-6 in the hTMΦ induced by PPM increased markedly too. And the induced expression effect of TNF-α and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTMΦ, and activate hTMΦ in vitro.     

An enzyme histochemical study of diaphragm in diabetic rats

AIM: The aim of this research is to study the earlier enzyme activity changes of the diaphragm in diabetic rats. METHODS: An enzyme histochemical method was used to observe the changes in the enzyme activities of dehydrogenases,hydrolases and oxidases in 4th week diabetic rat diaphragm. RESULTS: The activites of enzymes including SDH(Succinate dehydrogenase),MDH(Malate dehydrogenase), GDH(Glutamate dehydrogenase), ICDH(Isocitrate dehydrogenase), NADHD(NADH diaphorase), G-6-PD(Glucose-6-phosphate dehydrogenase), ACP(Acid phosphatase) and ANAE(Acid α-naphtyl acid esterase) were increased in diabetic diaphragm compared with the control. LDH (Lactate dehydrogenase)and CCO(Cytochrome oxidase) activities were decreased, whereas NADPHD(NADPH diaphorase) showed no changes in diabetic rats. Eleven kinds of enzyme were analysed with image analysis.Optical density (A) of SDH, MDH, GDH, ICDH, NADHD, G-6-PD, ACP and ANAE in diaphragm of diabetic rats were significantly higher than that of control rats (P<0.01). A value of LDH and CCO in diaphragm from diabetic rats were significantly lower than that of control rats (P<0.01). A value of NADPHD in diaphragm from diabetic rats showed no apparent alteration compared with the control rats(P>0.05). CONCLUSION: Increase in the aerobic capacity, decrease in the glycolytic capacity, and disturbance of lipid and energy metabolism were found in diaphragm of 4th week diabetic rats.     

Effects of icariin on expression of transforming growth factor β1 and type Ⅳ collagen in diabetic rat testis

AIM: To determine the beneficial effects of icariin on streptozotocin (STZ)-induced diabetic testopathy in rats. METHODS: The diabetic animal model was induced in male Sprague-Dawley rats by an injection of streptozotocin (40 mg/kg, iv). The rats were randomly divided into 3 groups: control group, model group and icariin (80 mg/kg, ig) group. Twelve weeks after injected with streptozotocin, all rats were anaesthetized and killed to remove the testes from scrotum. Serum concentrations of glucose and testosterone, and the levels of succinate dehydrogenase (SDH), acid phosphatase (ACP), γ-glutamyl transpeptidase (γ-GT) and lactate dehydrogenase (LDH) in testes were measured. The morphology of the testicular tissues was observed under light microscope. Immunohistochemistry was employed to determine the protein levels of TGF-β₁ and type Ⅳ collagen. RESULTS: Compared with control group, the content of serum glucose increased while the serum level of testosterone and the activitiy of SDH, ACP, γ-GT and LDH in testis decreased in model group (P<0.01). The histopathological examination showed that the diameters of seminiferous tubules and various grades of spermatocytes in the testis were markedly decreased. Compared with control group, the expression of TGF-β₁ and collagen Ⅳ was significantly increased in model group. These alterations were significantly attenuated in icariin group (P<0.01). CONCLUSION: Icariin evidently relieves testicular damage in rats with diabetic testopathy by improving the secretion of testosterone and reducing the expression of TGF-β₁ and collagen Ⅳ at protein level.     

Effects of cotransplantation of rat bone marrow mesenchymal ste m cells and bone marrow on hematopoiesis

AIM:To investigate effects of rBMMSC on he matopoiesis and immune reconstitution after allo-hematopoietic stem cells transp lantation (HSCT).METHODS:Allogeneic BMT model from Fischer344 rats (RT-1Al) to W istar rats (RT-1Au) was established.The effects of MSCs on hematopoietic recons titution and immune reconstitution were studied by observing the survival rate,peripheral blood counts,thymus counts,spleen counts,bone marrow counts and im mune function analysis at 30 days after transplantation.RESULTS:1.Cotransplantation of MSCs and bone marrow (BM) was d emonstrated to improve hematopoietic reconstitution.Lymphocyte and platelet cou nts in peripheral blood in cotransplantation groups were higher than those in co ntrol groups.More bone marrow neucleated cells were also observed in cotranspla ntation groups.2.Cotransplantation of MSCs and BM improved immune reconstituti on.First,overall thymic cellularity and spleen cellularity significantly incre ased in cotransplantation groups at day 30.Secondly,cotransplantation improved immune functional recovery.Non-specific lymphocytes proliferation reaction ind uced by ConA and LPS increased in cotransplantation group,and so did for alloge neic mixed-lymphocyte reaction.CONCLUSION:Hematopoietic reconstitution and immune reconstituti on were significantly enhanced by MSCs cotransplanted with BM.     

Effect of forsythiaside A on immune function in rats with ulcerative colitis

AIM To investigate the effect of forsythiaside A (FA) on immune function in rats with ulcerative colitis and its related mechanism. METHODS Healthy SD rats were randomly divided into 5 groups: control group (no treatment, normal feeding), model group (establishment of rat ulcerative colitis model), and low, medium and high doses of FA groups (treatment of the model rats with FA at 5 mg/kg, 20 mg/kg and 80 mg/kg, respectively). The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in rat colon tissues were measured by colorimetry, and the serum levels of tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2) and IL-4 were detected by ELISA. The spleen index and thymus index, the percentages of CD3⁺, CD4⁺ and CD8⁺ T-lymphocytes in peripheral blood mononuclear cells （PBMC）, the serum IgA and IgG levels, and the serum complement C3 and C4 levels were also determined. RESULTS The colon tissues of the rats in model group showed obvious inflammation and ulceration, indicating that the animal model was successfully established. Compared with model group, the colonic inflammation and ulceration were significantly attenuated in FA groups, among which the high dose had the best effect. Compared with control group, the spleen index and thymus index of the rars in model group were decreased (P<0.05), MDA content in colon tissues was increased (P<0.05), and SOD activity in colon tissues was decreased (P<0.05). The levels of CD3⁺, CD4⁺, CD8⁺ and CD4⁺/CD8⁺ T-lymphocytes in PBMC, and the serum levels of C3, C4 and IL-4 were decreased (P<0.05), while the serum levels of IgA, IgG, TNF-α, and IL-2 were increased in model group as compared with control group. Furthermore, the spleen index and thymus index of the rats in FA groups were increased (P<0.05), the MDA content in the colon tissues was decreased (P<0.05), and the SOD activity in the colon tissues was increased (P<0.05). The levels of CD3⁺, CD4⁺, CD8⁺ and CD4⁺/CD8⁺ T-lymphocytes in PBMC, and the serum levels of C3, C4 and IL-4 were increased (P<0.05), while serum IgA, IgG, TNF-α and IL-2 levels were decreased in FA groups as compared with model group (P<0.05). CONCLUSION Forsythiaside A effectively attenuates the colonic lesions in rats with ulcerative colitis, and its mechanism may be related to reinforcement of oxygen free radical scavenging power, alleviation of inflammatory response, and enhancement of immune function.     

Effect of Jiawei Sini Decoction on glucocorticoid receptor in thymocytes of chronic psychological stress rats

AIM: To observe the effect of Jiawei Sini Decoction (JWSND) on glucocorticoid receptor (GCR) in thymocytes of chronic psychological stress rats. METHODS: The rats were randomly divided into 4 groups, control group (C), model group (M), group treated by JWSND C₁, group treated by ginsenosides C₂. The number of thymocyte GCR sites and the GCR nuclear thanslocation rate were detected by radioimmunoassay. RESULTS: Compared with the control group, in the model group, the thymocyte weight index lowered significantly ( P<0.05 ), and the GCR nuclear thanslocation rate was increased significantly ( P<0.01 ), but the number of thymocyte GCR sites was unchanged. Compared with the model group, thymus gland weight indexes of C₁ and C₂ were increased significantly ( P<0.05 ), while the GCR nuclear thanslocation rate lowered significantly ( P<0.01 ). Moreover, no significant difference was found in all indexes between C₁ and control group. CONCLUSION: The inhibitory effect of glucocorticoid on the thymus could be significantly reversed by JWSND via suppressing the thanslocation of GCR from cytoplasm to nucleus in chronic psychological stress rats.     

Effect of fluvastatin on left ventricular remodeling after myocardial infarction in rats

AIM: To clarify the protective effect of long-term administration of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor fluvastatin on ventricular remodeling after myocardial infarction (MI) in rats and its mechanisms. METHODS: Myocardial infarction were established by ligated left coronary anterior artery in SD rats, 24 hours after the operation, the survival rats were treated by gavage fluvastatin (20 mg·kg^-1·d^-1) or distilled water for 8 weeks. Doppler echocardiography, homodynamic and cardiac histomorphometry were used to assess the ventricular remodeling and cardiac function. The plasma levels of total cholesterol (Tch), creatinine (Cr), glutamic-oxal (o) acetic transaminase (AST), lipid peroxidation (LPO), glutathione perioxidase (GSH-PX), nitrogen monoxide (NO₂^-/NO₃^-) were detected. RESULTS: The Tch, Cr and AST were not significant difference in groups. Left ventricular end-diastole pressure, right relative weight, left ventricular posterior wall thickness, collagen volume fraction and the lung weight were decreased in AMI+fluvastatin group compared to AMI group (P＜0.05, P<0.01); The levels of LPO, NO₂^-+NO₃^- in plasma and LPO in myocardium decreased, but plasma GSH-PX level increased in AMI+fluvastatin group (P＜0.05). CONCLUSION: Fluvastatin ameliorates the ventricular structural remodeling in a rat model of infarction, and delays the development of heart failure. The anti-oxidation mechanism of fluvastatin may take part in this process.     

Changes of gut mucosa antioxidant system and liver,renal function in rats after intestinal ischemia-reperfusion injury

AIM: To investigate the changes of the gut mucosa antioxidant system and liver, renal functions during rat intestinal ischemia-reperfusion injury. METHODS: 30 male Wistar rats underwent 45 min of intestinal ischemia by clamping the superior mesenteric artery followed by reperfusion. The levels of malondialdehyde (MDA), glutathione (GSH), the activities of antioxidant enzymes in the gut mucosa including catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), glutathione S- transferase (GST) activity and serum ALT, AST, BUN, Cr were assayed at 2, 4, 8, 12 and 24 h after reperfusion. RESULTS: The levels of MDA and GSH in the gut mucosa increased and decreased significantly at 2 h of reperfusion, respectively (P<0.05). MDA was still lower than sham at 24 h of reperfusion (P<0.05), while GSH decreased to 40% of sham at 4 h of reperfusion (P<0.01) but returned to the level of control at 12 h. The activities of CAT, SOD and GSH-Px did not show significant changes in rat after intestinal ischemia reperfusion. GST decreased 39% at 2 h of reperfusion compared with the sham group and decreased to 56% of sham at 4 h (P<0.05), but returned to the level of control at 12 h after reperfusion. Serum ALT, AST, BUN and Cr increased significantly at 2 h of reperfusion (P<0.05) and increased 208%, 100%, 103%, 41% compared with control at 4 h of reperfusion (P<0.01). However, at 24 h of reperfusion, they returned to normal. CONCLUSION: Intestinal ischemia/reperfusion diminishes GSH level and GST activity, increases MDA level and causes liver and renal reversible damages.     

Effect of NPPB and niflumic acid on expression of GCL and CLIC4 in H2O2 injured C6 glioma cells

AIM: To observe the effect of 5-nitro-2- (3-phenylpropylamino) benzoate acid (NPPB), niflumic acid (NFA) (chloride channel blockers) on malignant glioma C6 cells injured by hydrogen peroxide (H₂O₂). METHODS: The viability of C6 cells treated with NPPB, NFA and H₂O₂ was measured by MTT assay. LDH release rate and GSH contents were detected by ultraviolet spectrophotometry. mRNA levels of GCLC, GCLM and CLIC4 were determined by RT-PCR. CLIC4 protein level was detected by Western blotting. RESULTS: Compared to the control group, H₂O₂ treatment induced the decrease in cell viability and GSH contents, the increase in LDH release rate, the decrease in the expression of GCLC, GCLM and CLIC4 mRNA and the increase in CLIC4 protein level (P<0.05 ), respectively. Compared with the H₂O₂ group, H₂O₂ combined with NPPB or NFA treatment did not change the cell viability, the GSH contents and the GCLC, GCLM mRNA expression. However, the LDH release rate and CLIC4 protein level decreased (P<0.05). CONCLUSION: The chloride channel blockers NPPB or NFA lessen the oxidative injury of C6 cells through modulating the function of membrane and down-regulating the protein expression of CLIC4.     

Porcine surfactant in treatment of LPS-induced early-stage acute lung injury in rats

AIM: To evaluate the therapeutic efficacy of intratracheal instillation of porcine pulmonary surfactant (PPS) in rats with lipopolysaccharides (LPS)-induced early-stage ALI in this study.METHODS: SD rats weighing 200 g-300 g were randomly divided into 4 groups: LPS (1.5 mg·kg-1)+saline,LPS+PPS 100 mg·kg^-1,LPS+PPS 150 mg·kg^-1,LPS+PPS 200 mg·kg^-1.The PaO₂ and PaCO₂,as well as survival rate of rats were examined for 6 h after the start of PPS-instillation.Then,rats were killed and lungs were immediately removed for lung index (LI) and histological analysis.The bronchoalveolar lavage fluid (BALF) was collected for measurement of total protein (TP) contents,TNF-α level and white blood cell(WBC) numbers.RESULTS: Significantly increased PaO₂,reduced mortality rate,decreased total protein and TNF-α contents in BAL,as well as lung index and meliorated histological appearance were observed in three PPS-treated groups compared with group given saline after LPS (P<0.05).The therapeutic effect in PPS150 and PPS200 groups was better than that in PPS100 group.CONCLUSION: Intratracheal PPS instillation provides protective effect on acute lung injury in rats induced by LPS.     

Protective effect of ambroxol against the lung damage in chronically hypoxic rats

AIM: To investigate the effect of ambroxol on pulmonary and vascular injury in chronically hypoxic rats. METHODS: 36 male Wistar rats were randomly divided into 3 groups: normal control,chronically intermittent hypoxia(CIH) and ambroxol precaution group(AP).The CIH and AP groups were made into the chronically hypoxic models.The mean pulmonary artery pressure(PAPM) and the levels of plasma superoxide dismutase(SOD) and plasma nitric oxide(NO),lipid peroxide(LPO) were determined. The levels of the lung homogenates SOD, LPO, NO and the changes in pulmonary vascular structure were also examined. RESULTS: The levels of plasma and lung homogenates SOD,NO in CIH group were respectively significantly lower than that of normal control and AP group( P <0.01),but the levels of plasma and lung homogenates LPO were significantly higher( P< 0.01). PAPM in AP group is significantly lower than that of CIH group( P< 0.01);The damage of pulmonary artery smooth muscle cells and extra cell matrix of AP group is much slighter than that of CIH group. CONCLUSION: Ambroxol might be an effective protector in chronically hypoxic rats.     

Influence of Fu Fang Xiao Chai Hu Tang on level of interleukin-2 and value of CD4+/CD8+ in mice bearing Ehrlish ascites carcinoma

AIM: To investigate the effects of Fu Fang Xiao Chai Hu Tang (FFXCHT) on level of Interleukin-2 and value of CD4+/CD8+ in mice bearing Ehrlish ascites carcinoma(EAC).METHODS: The effects of FFXCHT on the EAC were observed and index of thymus and spleen were observed.The method of ［3H］-TdR incorporation was used to measure the IL-2 level,and the value of CD4+/CD8+ was assayed by ELITE calibur flow cytometry.RESULTS: Compared with the model group,FFXCHT inhibited the growth of EAC (P<0.01).The index of thymus and spleen of the model group were higher than that in the control group (P<0.05).Compared with the model group,FFXCHT increased the index of thymus and spleen,the IL-2 level and the value of CD4+/CD8+ (P<0.05),especially with the middle dose.CONCLUSION: The FFXCHT has antitumor effect.Increased IL-2 level and the value of CD4+/CD8+ play a regulatory role in the immunologic function in mice bearing EAC.     

Effect of astragalus polysaccharides on expression of MMP2 and MMP9 in annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats

AIM: To investigate the effect of astragalus polysaccharides (AP) on the expression of matrix metalloproteinase 2 (MMP2) and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats. METHODS: The model rats were randomly divided into model group (M group), and low-dose and high-dose AP treatment groups (L-AP and H-AP groups). The rats in sham operation group were used as negative control group (NC group). In addition, all the annulus fibrosus tissues were used for primary cell culture. Histological analysis was performed using HE staining and Safranin O staining. The expression of MMP2, MMP9, tissue inhibitor of metalloproteinase 2 (TIMP2) and collagen Ⅳ at mRNA and protein levels was analyzed by immunohistochemistry, Western blot and RT-qPCR. Cell-collagen adhesion assay was used to detect annulus fibrosus cell-collagen adhesion. RESULTS: The intervertebral discs of M group were degenerated, while astragalus polysaccharide improved the degenerative disc disease in the rats with cervical spondylosis. Compared with NC group, the expression of MMP2 and MMP9 in the annulus fibrosus tissues of M group increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression of MMP2 and MMP9 in L-AP group and H-AP group was significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). The cell-collagen adhesion in M group was significantly lower than that in NC group (P<0.05). Compared with M group, the cell-collagen adhesion in L-AP group and H-AP group was increased significantly (P<0.05). Compared with NC group, the expression of MMP2 and MMP9 in annulus fibrosus cells of M group was increased significantly, while the expression of TIMP2 and collagen Ⅳ was decreased significantly (P<0.05). Compared with M group, the expression levels of MMP2 and MMP9 in L-AP group and H-AP group of fibrocytes were significantly decreased, while the expression of TIMP2 and collagen Ⅳ was significantly increased (P<0.05). CONCLUSION: Astragalus polysaccharides inhibit the expression of MMP2 and MMP9 in the annulus fibrosus of cervical intervertebral discs from cervical spondylosis model rats and regulate the dynamic balance of MMPs and TIMPs in the extracellular matrix, thus inhibiting the degradation of collagen in the intervertebral disc matrix and having the potential research value for the treatment of intervertebral disc degeneration.     

Effects of aminoguanidine intervention on lens cell damage induced by D-galactose in rat eyes

AIM: To investigate the effects of aminoguanidine intervention on lens cell damage induced by D-galactose in rat eyes and its mechanism of action. METHODS: D-galactose (400 mg/kg) was injected into rats intraperitoneally for 14 weeks to induce the animal model of glycosylation and lens cell damage. Aminoguanidine (75 mg/kg, 150 mg/kg) were administered for 12 weeks by intragastric administration beginning at 3rd week. All animals were killed and blood samples were taken to measure the activity of aldose reductase, the level of fructosamine, the amounts of glycohaemoglobin and advanced glycation end-products. The lenses of eyes were taken to detect the activities of AR, GR, SOD and SDH. The amounts of AGEs, GSH, MDA or outleakage of LDH were measured, respectively. The ultrastructure and apoptosis of lens epithelial cells were examined by transmission electron microscope and flow cytometry, respectively. RESULTS: Animals were treated with D-galactose for 14 weeks, the serum level of fructosamine, the amounts of glycohaemoglobin and AGEs, and activity of AR were significantly increased. The amount of AGEs and activity of AR in lens were increased, the activity of antioxidase was decreased and oxidative product was increased. The apoptosis, the damages of mitochondria and cell nucleus in lens cells were observed. After treated with aminoguanidine for 12 weeks, the activity of AR and the level of fructosamine in serum, and the amounts of glycohaemoglobin and AGEs were significantly decreased (P<0.01). The outleakage of LDH and amount of MDA were also decreased (P<0.05, P<0.01), the activities of GR, SDH and SOD were increased (P<0.05, P<0.01). The apoptotic rate of lens cells was reduced (P<0.05, P<0.01). The morphology of mitochondria and cell nucleus were improved. CONCLUSION: D-galactose induces the damage of cell nucleus, the mitochondria and apoptosis in lens cells by glycosylation and oxidative stress. Aminoguanidine may supply the protective action through inhibiting the glycosylation and oxidative stress.