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Objective: To observe the effect of Pinelliae Rhizoma Praeparatum Cum Alumine polysaccharides(PRPCAP)on airway mucus secretion in rats with allergic asthma, in order to study the material basis of the "macromolecule" component of the polysaccharides as the original medicinal materials. Method: The 60 SPF-grade Wistar rats were induced by intraperitoneal injection of chicken ovalbumin (OVA) and aluminum-magnesium adjuvant, except for the normal control group. The OVA solution was aerosolized to establish a rat model of allergic asthma. After successful modeling, the rats were randomly divided into 5 groups, namely allergic asthma model group, positive drug group (montalurast sodium,5 mg·kg-1), high-dose PRPCAP group (400 mg·kg-1), middle-dose PRPCAP group (200 mg·kg-1) and low-dose PRPCAP group (100 mg·kg-1). The contents of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and bronchoalveolar lavage fluid (BALF) supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and the count of eosinophils (EOS) was detected by BALF sediment. The histopathological changes were observed by hematoxylin-eosin (HE) staining in lung tissue. The mRNA expression of mucin 5AC (MUC5AC) was detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR). Result: Compared with the normal control group, serum IL-4 level in the allergic asthma model group was significantly increased (Pγ level was significantly decreased (PPPPPγ in serum (PPPPConclusion: The "macromolecule" component of polysaccharides in the Pinelliae Rhizoma Praeparatum Cum Alumine may be the material basis for the efficacy of eliminating dampness and eliminating phlegm.
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Objective To observe the expression change of phosphorylation glogyen synthase kinase 3β at Ser9 [p-GSK3β (Ser9)] and investigate whether GSK3β involved in the retinal ganglion cell (RGC) and optic nerve in rat ocular hypertension model.Methods Thirty health Sprague Dawley (SD) rats with normal intraocular pressure (IOP) were randomly divided into 3 groups(10 cases in each group):control group,2 weeks ocular hypertension(OHT) group and 4 weeks OHT group.The ocular hypertension model was established by using episcleral veins ligation and cauterization.5 eyes from each group were taken for frozen section at week 2 and week 4 after establishing ocular hypertension model.The number of RGC was counted with Nissl's staining.The expression and distribution of p-GSK3β (Ser9) in RGC and optic nerve were detected by immunofluorescence staining.The retinal sample of other 5 rats was harvested for detecting the expression of total glogyen synthase kinase 3 β (total GSK3β) and p-GSK3β (Ser9) by Western blot.Results There was no statistical difference in IOP before modeling among three groups (P =0.89),but there was statistical difference after modeling (P ＜0.01),compared with the control group,the increased rate of IOP in 2 weeks OHT group and 4 weeks OHT group were 59.13％ and 26.93％ (all P ＜ 0.05).The average RGC numbers of 2 weeks OHT group and 4 weeks OHT group were 120 ± 10 per eye and 86 ± 7 per eye,which were both less than the number 149 ± 12 per eye in control rats,there were statistical differences (all P ＜ 0.05).Furthermore,the number of RGC in 4 weeks OHT group was less than 2 weeks OHT group (P ＜0.01).The expression of p-GSK3 β (Ser9) in the retina and optic nerve were positive detected by immunofluorescence,and the intensity of p-GSK3β (Ser9) immunofluorescence staining in RGC and optic nerve of OHT rats were less than model control eyes.The amount of p-GSK3β (Ser9) in retina of 2 weeks OHT rats decreased 19.89％ compared with control group rats (P ＜ 0.05),and the amount of p-GSK3β (Ser9) in retina of 4 weeks OHT rats was significantly decreased 36.46％ compared with control group rats(all P ＜ 0.05).While the total GSK3β did not change(P ＞ 0.05).Conclusion The number of RGC and the expression of p-GSK3β (Ser9) in RGC and optic nerve of OHT rat eyes decreased significantly.The change expression of the p-GSK3β (Ser9) in the RGCs and optic nerve may be correlated with RGC neurodegeneration in glaucomatous disorders.
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Objective To study the protective effect of vitamin C on UV-induced photoaged skin structure in rats and provide a basis for clinical medicine and health care.Methods Photoaging skin models were set up by chronic ultraviolet radiation in 24 SPF female Sprague-Dawley rats, irradiated twice weekly for 4 weeks.The rat photoaged skin was induced by exposure to a total dose of 123 J/cm2 UVA and 5 J/cm2 UVB for 4 weeks.Vitamin C was administered by gastric gavage in a dose of 50 mg/kg once daily for 30 days during the model development.We compared the pathological changes in the irradiated skin using HE, Masson’ s trichrome and Victoria blue B staining, and compared the ultrastructural changes by electron microscopy.Results Rat models with photoaged skin were developed successfully.The vitamin C group showed significant reduction of pathological severity including erythema, ulcers, epidermal cell proliferation, epidermal thickness, dermis vasodilation, inflammatory cell infiltration, fibroblast proliferation, endoplasmic reticulum dilatation, mitochondrial swelling and vacuolization, elastic fibers degeneration and focal accumulation, collagen fibers swelling with uneven thickness,compared with the rats of model group at the irradiation site.Conclusions Vitamin C is effective in reducing the structural damage of UV-induced photoaged skin in rats.
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Background Most animal models of experimental autoimmune uveitis (EAU) are single attacked procedure,with a different feature from the natural course of human recurrent autoimmune uveitis.So establishing a recurrent EAU model is necessary for the clinical study on EAU.Objective This study was to establish the recurrent EAU model in rat and investigate the ocular inflammation and pathological manifestation and interleukin-17 (IL-17)expression in the eye.Methods T cells isolated from the spleen and draining lymph nodes of Lewis rats immunized with interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide fragments (R16) 10 days earlier were re-stimulated with R16 in vitro and injected into naive syngeneic rats to establish the recurrent EAU models,and the normal Lewis rats were used as controls.The eyes of model rats were then examined daily for clinical signs of uveitis by slit-lamp biomicroscopy and scored Caspi's criteria.The rats were sacrificed 1 month,2,3months after injection respectively,and the retinal sections were prepared for the pathological examination by hemotoxylin & eosin staining.Immunohistochemistry was performed to detect the expression of IL-17 in the retina.Results Adoptive transfer of R16-specific T cells to Lewis rats induced recurrent uveitis.The inflammatory scores on the fourth day,the sixth day,and the inflammatory response disappeared on the tenth day after injection.However,the inflammatory reaction occurred repeatedly 4 or 5 times in the 2-month duration after that,and the right and left eyes of a single recipient showed a different pattern of relapse,and the clinical manifestations of EAU was similar to the natural course to those of human autoimmune uveitis.In the retinal specimens of 1-,2-and 3-month group,the number of inflammatory cells was gradually decreased as the time lapse.Compared with the normal group,the thicknesses of the entire retina,outer nuclear layer and inner nuclear layer decreased with a significant difference among the 4 groups (F=20.46,288.40,4.43,all P=0.00).The number of RGCs in the normal group,1-,2-and 3-month group was 231.27 ± 15.36,225.36 ± 17.79,132.18 ±9.39 and 67.45 ± 11.90,respectively,showing a significant difference among them (F=68.94,P=0.00).Immunohistochemistry showed that the scores of the IL-17 expression in the rat retina were 0.64 ± 0.17,1.92 ± 0.19,1.17 ± 0.23 and 0.83 ± 0.23,showing statistically significant difference (F=64.10,P=0.00).Conclusions The stimulation of R16-specific T cells can induce recurrent EAU in Lewis rat.Th17 is involved in the disease course.
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Objective To study the effect of Busui Jiannao Prescription on ethology, GSH content and GSH-PX activity in brain of PD model rats. Methods Forty-five the aging pure C57BL rats were divided into a negative injury group, a model group, and a Chinese medicine group. PD animal model were established by injection with MPTP and were treated with Busui Jiannao Prescription by intragastric administration. Score changes of PD model rats in the test of pole-climbing and suspending were observed. GSH content and GSH-PX activity were further measured. Results After treating with Busui Jiannao Prescription, the test scores in pole-climbing and suspending were improved, GSH content and GSH-PX activity in substantia nigra were increased. Conclusion Busui Jiannao Prescription can regulate the behavior coordination and repair the damage of dopaminergic neurons in the substantia nigra of PD model rats.
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A stable and reliable infected necrotizing pancreatitis (INP) model in rats was established in order to study the pathophysiological mechanism and pathological development rule of INP and explore the new therapeutic methods for the diseases. Forty-six SD rats were randomly divided into 5 groups. The animals in group A received the injection of 5% sodium taurocholate into the pancreatic duct and those in group B underwent that of E. Coli into the pancreatic duct. The rats in groups C, D and E were subjected to the injection of 5% sodium tanrocholate in combination with different con-centrations of E. Coli (103, 104, 105/mL, respectively) into the pancreatic duct. The dose of injection was 0.1 mL/100 g and the velocity of injection was 0.2 mL/min in all the 5 groups. Eight h after the injection, the survival rate of animals was recorded and the surviving rats were killed to determine the serum content of amylase and perform pathological examination and germ cultivation of the pancre-atic tissue. The results showed that acute necrotizing panereatitis model was induced by injection of 5% sodium taurocholate into the pancreatic duct. The positive rate of germ cultivation in group A was 12.5%. The acute necrotizing pancreatitis model was not induced by injection of E. Coli into the pan-creatic duct and the positive rate of germ cultivation in group B was 0. The INP model was estab-lished in groups C to E. The positive rate of germ cultivation was 60%, 100% and 100% and 8-h sur-vival rate 100%, 100% and 70% in groups C, D and E, respectively. It was concluded that a stable and reliable model of INP was established by injection of 5% sodium taurocholate in combination with 104/mL E. Coli into the pancreatic duct with a dose of 0.1 mL/100 g and a velocity of 0.2 mL/min. The pathogenesis of INP might he that the hemorrhage and necrosis of pancreatic tissue in-duced by sodium taurocholate results in weakness of pancreatic tissue in fighting against the germs.Meanwhile, the necrotic pancreatic tissue provides a good proliferative environment for the germs.
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Objective To study the effect of Tongbiding freezing-dryied powder for injection on the function of WDR neuron of spinal cord in living rats with chronic constriction injury (CCI),and clarify its analgesia mechanism in spinal cord center,and further explore whether it has the drug dependence at the cellular level. Methods Electricity physiology extracellular recording technique was used ,in the CCI living rat's expanded spinal cord waist stage the electricity activity of identical WDR neuron cells was recorded before and after administration of 20 mg?kg-1 Tongbiding. The electric discharge number of C response was observed before and 2,4,8,10 min after administration. The spontaneous electric discharge number. was observed before and 1,2,4,6 min after administration; wind-up phenomenon was also observed.Results The electric discharge number of C response was obtained in 8 WDR neurons,three were significant differences of the mean electric discharge number of C response between 2,4,8 min after administration and before administration (P
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Objective To establish a animal model for diffusion mesangioproliferation complicating focal segmentally glomerulosclerosis.Methods The models were established in the seventieth day by repetitive injection of 4mg/kg and 3mg/kg adriamycin to the SD rats in the 7th and 30th day after removal of their right kidneys.Results Serious edema, profuse albuminuria, hypoalbuminemia, hyperlipemia and renal failure were presented. Simultaneously, diffuse medium- or serious-grade proliferation of mesangial matrix, slight-grade proliferation of mesangial cells, segmental sclerosis of 22% of glomeruli, and overexpression of PDGF-B and bFGF in glomeruli were showed by pathologic examination, computerized image analysis and immunohistochemistry staining.Conclusions The models established in the transition stage from mesangioproliferation to glomerulosclerosis are helpful to study mechanisms and take measures for prevention and cure of chronic glomerulonephritis.
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Objective To investigate the mechanism of renal interstitial fibrosis after unilateral ureteral obstruction and renal protctive effect of angiotensin coverting enzyme inhibitor (ACEI),benazepril on rat kidney.Methods Twenty-four rats were randomly assigned to shame operation group (normal group),operation group, benazepril(10mg?kg -1 ?d -1 ) treatment group after unilateral ureteral obstruction. Renal tissues were examined by light and electronmicroscopy at the 2nd week after operation. Immunohistochemistry was applied to measure the expression of collagen Ⅲ,transforming growth factor ? 1 (TGF-? 1 ) and nuclear factor-kB(NF-kB) in the renal interstitium.Results In comparison with shame operation group, the expression of collagen Ⅲ , TGF-? 1 and NF-kB in the interstitium of operation group markedly increased (P
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Objective The polydiol, such as 1,2-propanediol, 1,3-propanediol,2,3-butanediol, was first used for the post-treatment of G-tBP,in order to find new fixative, for anticalcification, improving durability and clinical use. Method G-tBP with the polydiol post-treated were implanted sabcutaneously in back of weanling SD rats.60 days after the treatment,calcium concentration analysis,histologic examination,electron microscopy,heat shrink temperature and tissue humidity were tested.Results The polydiol-post treated bovine pericardium showed less calcification significantly as compared with the G-tBP after 60 days implantation(mean tissue clacium content(1 817?3 091) versus (176 32?43 27)?g/mg dry weight, P0 05). Conclusions ⑴G-tBP with the polydiol post-treated appears less calcification. ⑵The effect of polydiol post-treatment on anticalcification is correlated to the tanned temperature, at 25℃ is more effective than at 4℃.
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Objective To explore the molecular mechanism of glucocorticoid and high glucose-induced insulin resistance(IR).Methods Isolated rat adipocytes were cultured for 24h at 5 or 25 mM glucose with or without dexamethasone (Dex) 0 3?M. Then the glucose uptake , the phosphorylation of tyrosine of insulin receptor substrate (IRS)1/2 and expression of IRS 1/2 and protein kinase B(PKB) protein were measured by western blot analysis with molecular dynamics. Results These adipocytes treated with 25 mM glucose have shown to impair glucose uptake, IRS1 phosphorylation and the protein expression; Combined treatment with Dex enhanced high glucose-induced the suppression, and inhibited IRS1 tyrosine phosphorylation; High glucose increased IRS2 protein expression, Dex abolished partly its the effect. Conclusions High glucose can induce IR, Dex can aggravate the effect induced with high glucose. The mechanism may be involved in affecting the phosphorylation and expression of insulin signaling proteins.
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AIM: To investigate the protecting mechanisms of adjusting blood pressure and blood fat capsule(Radix angelicae sinesis,Rhizoma alismatis,poria,Rhizoma attactylodis macrocephalae,Rhizoma Chuanxiong,Radix Paeoniae alba,semen arecae,Folium perillae) in diabetic nephropathy rats induced by streptozotocin. METHODS: Forty-eight health male Wister rats were randomly divided into normal group,diabetic nephropathy group,adjusting blood pressure and blood fat capsule group and valstatan group.After experiment urine,blood,kidney samples were taken,and immunohistochemistry were performed. RESULTS: 24 hour urinary microalbumin,creatinine clearance,?_2-microglobulin,creatinine clearance ET1 and NOS in kidneys had distinct difference among three gruops. CONCLUSION: The provention and cure of DN ' function of adjusting blood pressure and blood fat capsule bears relationship with inhibition of ET-1,NOS's expression.