Effects of Yigu capsule on the level of cbf α-1 mRNA in bone of ovariectomized osteoporosis rats

AIM: To analyze the regulatory effect of Yigu capsule on core binding factor alpha 1 (cbf α-1) gene expression in bone of ovariectomized osteoporosis (OP) rats. METHODS: Thirty-six 10-month old Sprague-Dawley female rats were randomized into three groups: sham-operated group, model group and drug group. After intervention by the corresponding methods, the femurs were collected, SYBR green Ⅰ fluorescence quantitative PCR technique was applied with the internal control of GAPDH according to the relative quantitative formula (2-ΔΔCt), the differentially expressed multiples between the model group or drug group and sham-operated group were calculated. RESULTS: Quantitative formula analysis showed that the level of cbf α-1 gene expression in bone of model groups was decreased than that in sham-operated rats ［compared with sham-operated group, P<0.01, it was (9.9×105)-(1.6×104) times］. While in drug groups the level of cbf α-1 gene expression was 0.19 to 0.92 times than that in the sham-operated group, no significant difference was observed (P>0.05). CONCLUSION: The results of this study show that cbf α-1 gene expression in bone tissue of ovariectomized OP rat is decreased, and Yigu capsule increases the level of cbf α-1 gene expression in bone of OP, indicating that Yigu capsule induces bone marrow mesenchymal stem cells into osteoblasts.     

Effect of ACTH on pain behavior and the level of brain-derived neurotrophic factor and corticotropin-releasing hormone in the hippocampus of rats with chronic pain

AIM: To observe the effect of adrenocorticotropic hormone(ACTH) on the levels of brain-derived neurotrophic factor(BDNF), trkB and corticotropin-releasing hormone(CRH) in the hippocampus of arthritic rats.METHODS: The BDNF immunoreactivity(IR) and CRH-positive neurons were stained with immunohistochemistry and in situ hybridization methods, respectively.RESULTS: The BDNF-IR, CRH mRNA-positive neurons in the contralateral hippocampus of the arthritic rats were increased significantly, which was decreased markedly by intraperitoneal injection of ACTH. However, the effect of ACTH was attenuated after adrenalectomy(ADX).CONCLUSION: These results suggest that BDNF and CRH in the hippocampus of arthritic rats were involved in the modulation of chronic pain, ACTH produced its analgesic effect by inhibiting the increase in BDNF and CRF level. Adrenal is critical to the analgesic action of ACTH.     

Effects of exercise combined with forsamax on bone mineral density and nerve conduction in ovariectomy-induced osteoporosis rats

AIM: To observe the effects of exercise (EX) and forsamax (FOX) on bone mineral density (BMD) and nerve conduction in ovariectomized rats, and to determine whether exercise can enhance the effects of forsamax on postmenopausal osteoporosis (PMOP). METHODS: Ninety 6-month-old female SD rats were randomly divided into 2 groups: sham group (18 rats) and ovariectomized group (72 rats). Eight weeks after ovariectomy, dual-energy X-ray absorptiometry was applied to scan the BMD of the fourth lumbar vertebra. The level of serum estradiol (E₂) was analyzed by ELISA. The living ovariectomized rats were then divided into 4 groups: ovariectomized model group (OVX), fosamax treatment group (OVX+FOX), exercise treatment group (OVX+EX), and exercise combined with fosamax treatment group (OVX+FOX+EX). After treated with fosamax (1 mg/kg intragastrically) and/or exercise for 12 weeks, the BMD of the fourth lumbar vertebra, the motor nerve conduction velocity (MCV), motor distal latency (ML) and compound muscle action potential (CMAP) of the left femoral nerve were detected. The levels of serum type 1 procollagen carboxy-terminal propeptide (PICP) and type I collagen carboxy-terminal cross-linked telopeptide (ICTP) were also analyzed by ELISA. RESULTS: Twelve weeks after FOX and/or EX treatment, ovariectomized rats showed obviously lower BMD, higher PICP and ICTP than those in sham group (P<0.05). FOX or EX significantly increased BMD and reduced PICP induced by ovariectomy (P<0.05). FOX significantly reduced ICTP (P<0.05), but no remarkable difference was observed in OVX+FOX group as compared with OVX group. EX+FOX significantly increased BMD and reduced PICP and ICTP compared with FOX or EX alone (P<0.05). However, no obvious difference was observed between OVX+FOX group and OVX+EX group. No distinct difference in MCV and CMAP among the 5 groups was found. Neither ovariectomy nor FOX significantly affected ML. EX or EX+FOX made ML shorter than that in OVX group (fP<0.05), and ML was remarkably shortened in OVX+FOX+EX group than that in FOX alone group on the left femoral nerve (P<0.05). No significant difference between FOX and EX in the protective effects against ovariectomized rats was observed. CONCLUSION: Exercise and fosamax may restrain bone absorption effect of osteoclast and then improve the bone mineral density in ovariectomized rats.     

Effects of matrine on apoptosis and related protein expression in human medulloblastoma D341 cells

AIM: To investigate matrine-induced apoptosis of human medulloblastoma D341 cells and the expression of Bax, Bcl-2, serine/threonine kinase Akt and phosphorylated Akt (p-Akt) in vitro. METHODS: D341 cells were divided into experimental groups (added with matrine at different concentrations) and control group (under the same conditions without matrine). The proliferation of D341 cells was analyzed by CCK-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining and the expression of Bax, Bcl-2, Akt and p-Akt was detected by Western blotting. RESULTS: Matrine significantly inhibited the proliferation of D341 cells and increased the apoptosis in a dose- and time-dependent manner. The cell apoptosis was characterized by chromatin condensation with margination of chromatin to the nuclear membrane, increased when and larger cytoplasmic vacuoles, and formation of apoptotic body after treatment with matrine. The expression of Bax increased, while the expression of Bcl-2 and p-Akt decreased when the drug concentration gradually increased. CONCLUSION: Matrine induces the apoptosis of human medulloblastoma D341 cells in vitro by activation of Bax, down-regulation of Bcl-2 and reduction of p-Akt expression level in the PI3K/Akt signaling pathway.     

Effects of the juice of Fructus Hippophae (JFH) on thrombocytopenia induced by cyclophosphamide in rats

AIM:To investigate the effects of the juice of fructus hippophae(JFH) on thrombocytopenia in rats.METHODS:Forty-eight wistar rats were divided into control group, model group, Yixuesheng-Jiaonang (YSJN) group and JFH group. Rats were injected intraperitoneally with cyclophosphamide (CTX, 30 mg/kg) or saline once a day for 3 days. And then, Saline, YSJN or JFH was administered (ig) once a day for 11 days. On the second, fouth, sixth and eighth day, the thrombocyte were counted, the mean platelet volume (MPV) and PDW were examined. On the eighth day, the cAMP, cGMP in platelets and platelet aggregation function were also examined.RESULTS:JFH could shorten the time of blood coagulation, increase the count of platelet, and enhance the platelet aggregation function. The experiment also showed that the JFH could decrease cGMP content in platelet.CONCLUSION:JFH could enhance blood coagulation and quality of platelet and prevent thrombocytopenia induced by cyclophosphamide in rats.     

Effect of tenuigenin on tau protein phosphorylation at Ser396 site in neurons of AD rats induced by Aβ1-40

AIM:To investigate the effect of tenuigenin(TEN) on hyperphosphorylation of tau protein in neurons of amyloid β-peptide_1-40(Aβ_1-40) -induced Alzheimer disease(AD) rats. METHODS:Aβ_1-40 was injected into hippocampus CA1 region of the rats to establish AD model. TEN at different doses(18.5 mg/kg, 37.0 mg/kg and 74.0 mg/kg) was intragastrically administered. The protein expression of protein kinase A(PKA),protein phosphatase 2A(PP2A), total tau and p-tau(Ser³⁹⁶) in the neurons was observed by the method of immunohistochemistry. The protein content of total tau and p-tau(Ser³⁹⁶), and the expression level of PKA and PP-2A were detected by Western blotting analysis. RESULTS:In Aβ_1-40 group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were significantly increased compared with those in sham operation group. Meanwhile, the expression of PP2A in Aβ_1-40 group was lower than that in sham operation group. In TEN treatment group, the level of total tau, the phosphorylation of tau protein and the expression of PKA were markedly decreased, and the expression of PP2A was increased as compared with Aβ_1-40 group. CONCLUSION:TEN may protect the neurons from the toxic effect of Aβ_1-40 and reduce the hyperphosphorylation of tau(Ser³⁹⁶) in the neurons of AD rats by activating the expression of PP2A and inhibiting the expression of PKA.     

Effects of astragaloside on the levels of sex hormone and oxidative stress injury in rats with polycystic ovary syndrome

AIM To investigate the effects of astragaloside on the levels of sex hormone and oxidative stress in rats with polycystic ovary syndrome （PCOS）. METHODS Female SD rats （n=60） were randomly divided into normal control group, model group, Diane-35 （0.339 2 mg/kg） group, low dose astragaloside （12.5 mg/kg） group and high dose astragaloside （50 mg/kg） group, with 12 rats in each group. The PCOS model was induced by letrozole （1 mg/kg）, which was administered by gavage once a day for 3 weeks. After administration, the estrus cycle of the rats was observed by vaginal smear, and the ovarian index was calculated. HE staining was used to observe the histopathological changes of the ovaries. Serum levels of the sex hormones testosterone （T）, estradiol （E₂）, luteinizing hormone （LH） and follicle-stimulating hormone （FSH） were measured by ELISA. The levels of superoxide dismutase （SOD）, malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） in serum and ovarian tissue were detected by colorimetry, and the protein levels of steroidogenetic acute regulatory protein （StAR） and apoptosis-related proteins cleaved caspase-3, Bax and Bcl-2 in ovarian tissue were detected by Western blot. RESULT Compared with control group, the oestrous cycle of the rats in model group was disorder, and the ovarian index was increased, ovary was polycystic. The serum levels of T, LH and MDAwere significantly increased （P<0.05）, while the contents of E2, FSH and the activities of GSH-Px and SOD were significantly decreased （P<0.05）. The levels of MDA, StAR, cleaved caspase-3 and Bax proteins in ovarian tissue were significantly up-regulated （P<0.05）. GSH-Px and SOD activities and Bcl-2 protein levels were significantly down-regulated （P<0.05）. CONCLUSION Astragalosideeffectively balances the levels of sex hormone in PCOS rats and relieves the oxidative stress injury, the mechanism may be related to the inhibition of StAR expression.     

高温强光胁迫对砂梨叶片光合作用、D1蛋白和Deg1蛋白酶的影响

【目的】研究夏季高温强光下砂梨叶片光合特征的变化,探讨梨光抑制的分子机制。【方法】以1年生‘翠冠’和‘圆黄’梨的盆栽苗为试验材料,测定了夏季晴天高温强光对气体交换参数和光响应曲线的影响,以及D1蛋白、Deg1蛋白酶含量和PsbA mRNA表达的变化。【结果】‘翠冠’和‘圆黄’梨叶片在夏季晴天中午高温强光的双重胁迫下,净光合速率(Pn)、气孔导度(Gs)、蒸腾速率(Tr)、表观量子效率(AQY)、D1蛋白和Deg1蛋白酶含量显著下降,而PsbA mRNA积累增加。到了17:00,‘翠冠’的D1蛋白、Deg1蛋白酶含量得到恢复,Pn略有回升,而‘圆黄’未出现明显的恢复迹象。【结论】高温强光下2个梨品种的叶片均发生了光抑制,低含量的Deg1不是降解D1蛋白的主要蛋白酶。高温强光诱导的D1蛋白净含量的减少主要是由于D1蛋白合成在翻译阶段受到了抑制。从下午的光合和D1蛋白的恢复来看,‘翠冠’梨的PSII比‘圆黄’梨更抗高温。     

Effects of drynaria total flavonoids on serum levels of leptin,interleukin 6, prostaglandin E2 and expression of bone β2-adrenergic receptor in ovariectomized osteoporosis rats

AIM: To investigate the effects of drynaria total flavonoids on serum levels of leptin (LEP), interleukin 6(IL-6), prostaglandin E₂(PGE₂) and the expression of bone β₂-adrenergic receptor (ADRB2) in a rat model of ovariectomized osteoporosis(OP). METHODS: The osteoporosis model was established by ovariectomy. Twelve weeks after modeling,bone mineral density (BMD) was determined by dual-energy X-ray absorptiometry to verify successful modeling.Enzyme-linked immunosorbent assay was applied to detect the concentrations of LEP, IL-6 and PGE₂ in serum. The expression of ADRB2 was determined by immunohistochemical technique. RESULTS: Compared with sham group,BMD of the rats in model group significantly decreased in multiple regions 12 weeks after modeling(P<0.01). The serum levels of LEP, IL-6 and PGE₂ in model group were significantly higher than those in sham group(P<0.05). The levels of LEP, IL-6 and PGE₂ in drynaria total flavonoids group were significantly lower than those in model group(P<0.01). No significant difference of PGE₂ between these 2 groups was observed. The ADRB2 expression in sham group and treatment group was significantly different from that in model group, and no significant difference between sham group and treatment group was found. CONCLUSION: The serum levels of LEP, IL-6 and PGE₂ and the expression of bone ADRB2 increased in OP rats.Drynaria total flavonoids reduce the production of LEP, IL-6 and the expression of ADRB2, and suppress the bone absorption, which may be one of the mechanisms in treating OP.     

Effect of Aima recipe on the protein and mRNA expression of TNFα and ICAM-1 in the bronchus of rats with chronic bronchitis

AIM:To study the effect of Aima recipe (AM), a traditional Chinese medicine, on the protein and mRNA expression of TNFα and ICAM-1 in the bronchus of rats with chronic bronchitis(CB).METHODS:15 male Wistar rats were randomly divided into three groups: control group, chronic bronchitis (CB) group, CB plus AM group. The protein and mRNA expression of TNFα and ICAM-1 were assayed by immunohistochemistry and in situ hybridization methods.RESULTS:TNFα, ICAM-1 protein and mRNA were more strongly expressed in the area of bronchial epithelium in the CB group compared with control group (P＜0.01), treatment with AM significantly decreased their expressions(P＜0.05). CONCLUSION:Downregulating the expressions of TNFα and ICAM-1 mRNA and protein in the area of bronchial epithelium may be one of the mechanisms underlying therapeutic effect of AM on chronic bronchitis.     

Effects of gypenosides on cell cycle-related protein expression and calcium homeostasis in hippocampal neurons of rats treated with β-amyloid protein fragmant 1-40

AIM：To observe the effects of gypenosides on the expression of cell cycle-related protein and calcium homeostasis in the hippocampal neurons in the animal model of Alzheimer’s disease (AD) induced by Aβ₁-40.METHODS：The animal model of AD was established through injection of Aβ₁-40 into hippocampus in rats.The ability of learning and memory were verified by the performance of Y-shape maze task.With the method of immunohistochemical staining coupled integral absorbance analysis,the expression of cell cycle related protein in hippocampus was determined in rats.The contents of cytoplasm Ca2+ were determined using Fura-2/AM-fluorescence method.The effects of gypenosides on above indices were studied.RESULTS：Compared with the controls,the learning and memory ability of Aβ₁-40-injected rats were lower,the expression of cyclin A and cyclin B₁ protein in rat hippocampus neurons had hoisted,the contents of calcium in hippocampus were increased significantly.Gypenosides,however,improved all the indexes in varying degrees.CONCLUSION：These results imply that gypenosides improves the learning and memory ability of the rats treated with Aβ₁-40,reverses the expression of cell cycle-related protein and maintains the calcium homeostasis in hippocampus neurons.     

Effect of glucagon-like peptide-1(7-36)NH2 on insulin secretion and membrane potentials of pancreatic islet β-cells

AIM: To observe pathomorphological changes in cerebral cortex and hippocampus in the mouse with synthetic vascular dementia. METHODS: The synthetic vascular dementia model was produced in the mouse. Animals were killed 7 d, 15 d, and 30 d after the operation, brain tissues were removed and embedded in paraffin. Section of 8μm thickness were stained with hematoxylin-eosin(HE) and Nissl methods, and observed with light microscope. RESULTS: The cerebral cortex in the mouse became thinner on the seventh day, karyopyknosis in partial nervous cells was formed, the number of local neurons was reduced, sieve structure was observed, and glial cells proliferated, with the similar results 15 d and 30 d after operation. Model mouses hippocampal cells in CA₁ area were reduced and almost disappeared 30 d after operation. At the same time, glial cells were abundantly proliferated, tubercles were formed. Cells in CA₂, CA₃ area were also reduced and hippocampal sclerosis occurred. CONCLUSION: Delayed necrosis of hippocampal pyramidal cells may be the pathological basis of ischemia cerebral vascular dementia.     

GA_3和PP_(333)对苹果花芽形态建成及其内源激素比例变化的影响

以红富士、首红苹果为研究对象,连续3年用PP333(多效唑)1000mg/L和GA3(赤霉酸)1000mg/L处理,结果表明:PP333可使叶芽的节位数增长,但对花芽的节位数没有明显的影响。而GA3对叶芽节位数没有影响,但首红苹果花芽节位数有减少趋向。二者处理对苹果花芽形态分化开始时期没有影响,但PP333加速了花芽形态分化进程,GA3延迟了花芽形态分化进程。喷PP333提高了ZR(玉米素核苷)/IAA(吲哚乙酸),ZR/GAS(赤霉素)、ABA(脱落酸)/IAA和ABA/GAS比值,从而促进了花芽形成。相反,GA3处理降低了ZR/IAA、ZR/GAS、ABA/IAA和ABA/GAS比值,而抑制了花芽形成。     

Correlation between the effect of angiotensin-(1-7) on cardiac hypertrophy and extracellular signal-regulated kinase in pressure-overloaded rats

AIM: To investigate the effects and mechanisms of angiotensin-(1-7) on cardiac hypertrophy in pressure-overloaded rats. METHODS: Ar at model of pressure-overloaded heart was induced by constriction of abdominal aorta. Seventy-five male Sprague Dawley rats were randomized to sham-operated group, model control group and angiotensin-(1-7) treatment group. They were treated with intravenous infusion of angiotensin-(1-7) (25 microgram/kg per hour) or saline by minipump. RESULTS: Abdominal aortic banding resulted in a significant increases in LVW/BW, myocardial angiotensinⅡlevels, and p-ERK1/2 expression. Angiotensin-(1-7) had no effect on aortic banding-induced increases in myocardial angiotensinⅡlevels, but it significantly attenuated aortic banding-induced increases in LVW/BW and p-ERK1/2 expression. CONCLUSION: Angiotensin-(1-7) attenuates the development of cardiac hypertrophy in pressure-overloaded rats. It may be associated with the inhibition of p-ERK1/2 expression in cardiac tissue.     

Effects of perindopril at different doses on cardiac function and ACE2/Ang-(1-9)/Ang-(1-7) axis of ischemic cardiac dysfunction rabbits

AIM: To investigate the different dose of perindopril on cardiac function in the rabbits with ischemic cardiac dysfunction. METHODS: Male rabbits weighing 2.5~3.0 kg(n=30) were randomly divided into 3 groups(n=10):high dose perindopril group(HD group), low dose perindopril group(LD group) and cardiac dysfunction group(CD group). The Left anterior descending coronary artery of the rabbits was ligatured for model preparation. In HD group, the rabbits were treated with perindopril split normal saline solution(1 g/L)2 mL·kg^-1·d^-1. In LD group, the rabbits were treated with perindopril split normal saline solution(0.33 g/L)2 mL·kg^-1·d^-1. In CD group, the rabbits were treated with normal saline solution 2 mL·kg^-1·d^-1. Four weeks after treatment, the cardiac function was measured via echocardiography, the mRNA expression of angiotensin-converting enzyme 2(ACE2) and angiotensin type 2 receptor(AT2R) was analyzed by real-time PCR, serum angiotensin(Ang)-(1-9) and Ang-(1-7) levels were detected by ELISA. RESULTS: Compared with CD group, the cardiac function of the 2 groups treated with perindopril was significantly improved(P<0.01), and more improvement in HD group was observed than LD group(P<0.05). The serum angiotensin(Ang)-(1-9) and Ang-(1-7) level and the mRNA expression of ACE2 and AT2R in the 2 groups treated with perindopril were significantly improved(P<0.01). Compared with LD group, the mRNA expression of ACE2 and AT2R and the serum levels of Ang-(1-9) in HD group were significant improved(P<0.05), while no difference of serum Ang-(1-7) level was observed. Correlation analysis revealed that the improvement of the cardiac function was associated with serum Ang-(1-9) level, mRNA expression of ACE2 and AT2R(P<0.01), but has no significant correlation with serum Ang-(1-7) level. CONCLUSION: High dose of perindopril may improve more cardiac function in ischemic cardiac dysfunction model in rabbits. The mechanism may relate to increasing serum Ang-(1-7) level to activate AT2R.     

Effects of angiotensin-(1-7) on calcification in vascular smooth muscle cells and its signal transduction pathway

AIM: To clarify the effects of angiotensin-(1-7) on the calcification in rat vascular smooth muscle cells(VSMCs) and its signal transduction.METHODS: Calcification of cultured rat VSMCs was prepared by incubation of VSMCs with β-glycerophosphate.Calcification was confirmed by Von Kossa staining.The cells were treated with angiotensin-(1-7).The calcium content,alkaline phosphatases activity,osteocalcin and Cbfa1 mRNA expression were also measured.RESULTS: Angiotensin-(1-7) inhibited the increases of calcium content,alkaline phosphatases activity（P>0.05）,osteocalcin concentration and Cbfa1 mRNA expression in calcified VSMCs（P<0.05）,and the effects of angiotensin II on calcium content,alkaline phosphatases activity,osteocalcin concentration and Cbfa1 mRNA expression in calcified VSMCs were also inhibited （P<0.05）.Angiotensin-(1-7) increased cAMP concentration in calcified VSMCs（P<0.05）and selective PKA inhibitor blocked the effects of angiotensin-(1-7) on calcium content,alkaline phosphatases activity,osteocalcin concentration and Cbfa1 mRNA expression（P<0.05）.CONCLUSION: Angiotensin-(1-7) can inhibit beta-glycerophosphate-induced calcification in VSMCs through cAMP-PKA-Cbfa1 pathway and antagonize the effect of angiotensin II on calcification in VSMCs.     

Effects of Bene Jones protein and TGF-β1 on the proliferation of rat renal proximal tubular cells in vitro

AIM:To study the effect of Bene Jones protein (BJP) from multiple myeloma(MM) patient and TGF-β₁ on cultured renal proximal tubular cell(PTC) proliferation.METHODS:[H³]TdR incorporation was used to study the effect of λBJP and TGF-β₁ on cultured rat NRK.52E PTC proliferation, the expression of TGF-β₁ in the supernatant of PTC cultured with BJP was assessed with ELISA.RESULTS:① [H³]TdR incorporation of PTC was inhibited by BJP in a dose-dependent manner, when co-cultured with 100-800 μmol/L BJP and 2.0 μg/L TGF-β₁, the [H³]TdR incorporation was lower than that of BJP alone, especially when BJP≥400 μmol/L;②The expression of TGF-β₁ in the supernatant of PTC cultured with BJP was increased, especially when BJP≥400 μmol/L(P＜0.05);③ The [H³]TdR incorporation of PTC was also inhibited by exogenous TGF-β₁ in a dose-dependent manner.CONCLUSION:λBJP has antiproliferative effect on rat PTC in vitro, The effect is related with stimulating the PTC to produce excessive TGF-β₁, which also has antiproliferative effect on PTC in some degree.