Therapeutic effect of MCP-1 mediated macrophages on ovarian epithelial carcinoma

AIM: To investigate the therapeutic effect of MCP-1 mediated macrophages on ovarian epithelial carcinoma and its mechanisms.METHODS: Retorviral expression vectors pLXSN/MCP-1 was transfected into the packaging cell line PA317 by lipofectin-mediated gene transfer system. The virus particles containing MCP-1 gene were collected to infect NuTu-19. RT-PCR and Boyden Chamber were used to confirm the expression of MCP-1. Rat Fischer344 spleen macrophages were isolated. MTT method was applied to investigate the tumoricidal effect of macrophages. The survival time of the intraperitoneal disseminating ovarian cancer animal model was observed, and flow cytometry method was applied to analyze the expression of CD25 or CD44v6, and then the anti-tumor mechanisms of gene modified tumor cell lines were discussed. RESULTS: Stable MCP-1 expression in the cell line NuTu-19/MCP-1 possessed the chemotatic activity. The maximum killing ratio of macrophages on NuTu-19/MCP-1 cells was 28%. In the animal models immunized by MCP-1 expressing cells, prolonged survival time was showed which had statistical significance compared with that in the control group (P<0.05). The expression rate of CD25 (25.82%) in the NuTu-19/MCP-1 cells was higher than that in NuTu-19/neo cells (8.73%). The expression of CD44v6 in NuTu-19/MCP-1 cells was significantly lower than that in control NuTu-19/neo cells. CONCLUSION: MCP-1 mediates macrophages and suppresses the growth of NuTu-19. MCP-1 gene modified tumor cells can induce anti-tumor immunity. This strategy would be used as a promising approach for the treatment of ovarian cancer.     

Effects of Xijiao Dihuang and Yinqiao San decoction on ICAM-1 and VCAM-1 expression in mouse lung tissues and rat pulmonary microvascular endothelial cells infected with influenza virus

AIM：To investigate the effects of Xijiao Dihuang and Yinqiao San decoction (XDY) on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in mouse lung tissues and rat pulmonary microvascular endothelial cells (RPMVECs) infected with influenza virus, and to explore its mechanism for treatment of viral pneumonia. METHODS：Fifty-four male BALB/c mice were randomly divided into normal group, model group and XDY group (n=18 in each group). The viral pneumonia model was established by intranasally dripping influenza A (H1N1) virus into the mice. The mice in XDY group were treated with XDY 1 h after dripping the virus. The expression of ICAM-1 and VCAM-1 in lung tissues was examined by immunohistochemical staining 2, 4 and 6 d after infection. On the other hand, RPMVECs were obtained from male Wistar rats and primarily cultured. The cells were randomly divided into control group, virus group, virus+XDY group, tumor necrosis factor α (TNF-α) group and TNF-α+XDY group. The mRNA and protein expression of ICAM-1 and VCAM-1 was evaluated by real-time PCR and flow cytometry 24 h after infection. RESULTS：Virus exposure increased ICAM-1 and VCAM-1 expression in mouse lung tissues (P<0.01), and XDY treatment attenuated this effect (P<0.01). Virus and TNF-α both led to the increases in mRNA and protein expression of ICAM-1 and VCAM-1 in RPMVECs (P<0.01), which were also reduced by treatment with XDY (P<0.01). CONCLUSION：Treatment with XDY decreases virus-induced ICAM-1 and VCAM-1 expression, suggesting an important role of XDY in treatment of viral pneumonia.     

Expression of connexin 40 and connexin 45 in renal tissue of rats with chronic renal failure and effect of treatment with Astragalus polysaccharide and stachydrine combination

AIM：To explore the expression of connexin 40 (Cx40) and connexin 45 (Cx45) in chronic renal failure rats, and to investigate the effect of an Astragalus polysaccharide and stachydrine combination on the expressions of the 2 proteins, and the treatment effect of the combination on chronic renal failure. METHODS：Wistar rats were randomly divided into 4 groups:control group (6 rats), chronic renal failure model group (8 rats), low-dose Astragalus polysaccharide (200 mg·kg-1·d-1) and stachydrine (2.5 mg·kg-1·d-1) combination group (8 rats), and high-dose Astragalus polysaccharide (400 mg·kg-1·d-1) and stachydrine (5 mg·kg-1·d-1) combination group (8 rats). The rat model of chronic renal failure was induced by adenine (200 mg·kg-1·d-1) gavage. After 7-week administration, the expression of Cx40 and Cx45 in the renal tissue was determined by immunohistochemical staining. The renal function, electrolyte levels, and pathological changes of the kidneys were also analyzed. RESULTS：The expression of Cx40 and Cx45 was increased in the renal tissue of chronic renal failure rats. The combination of Astragalus polysaccharide and stachydrine reduced the kidney weight and index, attenuated the pathological renal damage in the model rats, decreased the expression of Cx40 and Cx45, maintained the stability of serum electrolytes, and decreased the levels of serum creatinine and blood urea nitrogen. CONCLUSION:Chronic renal failure caused the increase in the expression of Cx40 and Cx45 in the rat renal tissue. The combination of Astragalus polysaccharide and stachydrine reduces the expression of Cx40 and Cx45 in the renal tissue, and improves the renal function, thus playing a role in protecting the kidneys.     

Effects of antihistamine treatment on mast cell infiltration and c-Kit and SCF expression in liver tissues of rats with experimental hepatitis

AIM：To study the infiltration of mast cells and the expression of c-Kit and stem cell factor (SCF) in liver tissues of rats with experimental hepatitis and their changes after antihistamine (AH) treatment. METHODS：Thirty Wistar rats were divided into 3 groups at random: normal control (NC) group, chronic hepatitis (CH) group and AH group. The rat model of CH was established by composite factors (subcutaneous injection of carbon tetrachloride, accompanied by a diet containing high cholesterol, high alcohol, low protein and low choline). The rats in AH group were treated with ketotifen based on CH. At the end of the 4th week, blood samples were taken to determine plasma tryptase (TS) and histamine (HA) levels. Liver tissues were taken to detect HA content, observe the histological changes with HE staining and count the number of mast cells with toluidine blue (TB) staining. The mRNA and protein expression of c-Kit and SCF in liver tissues was detected by RT-PCR and immunohistochemistry. RESULTS：(1) The plasma TS and HA levels and liver HA content in CH group were significantly increased compared with NC group (P<0.05), while those in AH group were obviously decreased compared with CH group (P<0.05). (2) Fatty degeneration and fibrosis were observed in CH group under light microscope, but the hepatic injury was obviously attenuated in AH group. TB staining showed there were many degranulating and degranulated mast cells filled with purple granules around liver blood vessels and in fiber interval in CH group, and there were few purple granules in the cytoplasm of mast cells in AH group. The number of mast cells in CH group was increased compared with NC group (P<0.05), and that in AH group was reduced compared with CH group (P<0.05). (3) The results of RT-PCR showed that AH down-regulated the expression of c-Kit and SCF mRNA (P<0.05). The expression of c-Kit and SCF proteins in liver tissues increased in CH rats (P<0.05 vs NC group), decreased after AH treatment (P<0.05 vs CH group) and was positively correlated with liver HA content (P<0.05). CONCLUSION：These data suggest that an inflammatory pathway mediated by mast cell activation is involved in experimental hepatitis. Ketotifen can reduce mast cell degranulation by down-regulating the expression of mast cell membrane receptor c-Kit and its ligand SCF, thereby attenuating the liver inflammation.     

珊瑚状猴头菌多糖对大鼠肝抗氧化及代谢调节

探讨珊瑚状猴头菌多糖的抗氧化作用和对大鼠肝脏代谢的相关基因表达的影响。将大鼠随机分为正常对照组、多糖干预组和模型组。对照组喂饲基础饲料,多糖干预组（剂量1组、剂量2组）和模型组喂饲高胆固醇饲料,同时多糖干预组（剂量1组、剂量2组）给予多糖灌胃5周,5周后处死大鼠取肝脏。分别测定大鼠肝脏的丙二醛（MDA）含量,总超氧化物歧化酶（T-SOD）、谷胱甘肽过氧化物酶（GSH-Px）、过氧化氢酶（CAT）的活力。使用实时荧光定量PCR分别检测大鼠肝脏相关基因表达量：酰辅酶A：胆固醇酰基转移酶2（ACAT2）、卵磷脂胆固醇酰基转移酶（LCAT）。与模型组比较：多糖剂量1组、剂量2组大鼠肝脏的MDA含量显著下降（P〈0．01）,T-SOD活力显著升高（P〈0．01）,GSH-Px活力显著升高（P〈0．01）,CAT活力显著提高（P〈0．01）;多糖剂量1组、多糖剂量2组大鼠肝脏的基因表达量ACAT2mRNA显著下降（P〈0．01）;LCATmRNA显著升高（P〈0．01）。珊瑚状猴头菌多糖具有显著的抗氧化作用,对大鼠肝脏的代谢具有调节作用。     

Influence of high-mobility group box 1 on proliferation of neural stem cell in pre-infarction cortex of focal cerebral ischemia/reperfusion model rats

AIM：To investigate the influence of high-mobility group box 1 (HMGB1) on the proliferation of neural stem cells in peri-infarction cortex of focal cerebral ischemia/reperfusion model rats. METHODS：Male SD rats (n=48) were randomly divided into sham group, ischemia/reperfusion (I/R) group, RNA interference group and negative interference group. The rat middle cerebral artery was blocked to establish focal cerebral I/R model (ischemia for 1 h and reperfusion for 7 d). Lentivirus vector of HMGB1 shRNA was used to suppress the HMGB1 protein expression in the rat brain. The effect of RNA interference was evaluated by the methods of double-immunofluorescence labeling of HMGB1/GFAP and Western blotting. The proliferation of neural stem cells in the peri-infarction cortex was assessed by double labeling of BrdU/nestin. RESULTS：The protein expression of HMGB1 in I/R group was much higher than those in sham group (P<0.05). RNA interference effectively inhibited the HMGB1 expression (P<0.05). Double labeled BrdU/nestin positive cells in I/R group were more than that in sham group (P<0.05). The double labeled BrdU/nestin positive cells were significantly decreased in RNA interference group (P<0.05). CONCLUSION：Focal cerebral ischemia/reperfusion injury promotes the proliferation of neural stem cells in peri-infarction cortex by increasing HMGB1 protein level.     

Effects of angiotensin II-induced hypertension chemotherapy on the activity of interleukin-1β converting enzyme in transplanted intracerebral rat gliomas

AIM: To investigate the activity of interleukin-1β converting enzyme in transplanted intracerebral rat gliomas under angiotensin II-induced hypertension chemotherapy. METHODS: The brain tumor model was produced in Wistar rats by stereotaxic inoculation of C6 glioma cells (1×10¹² /L). Tumor-bearing rats were treated with carmustine, teniposide and lisplatin (chemotherapy) during angiotensin II-induced hypertension. Then, the survival time of tumor-bearing rats, tumor blood flow, the concentration of drug, volume of gliomas and the activity of interleukin-1β converting enzyme in glioma were examined.RESULTS: The survival time of tumor-bearing rats was significantly longer in chemotherapy with angiotensin II-induced hypertension group than that of chemotherapy alone. In addition, regional tumor blood flow, the concentration of chemotherapeutic drug and the activity of interleukin -1β converting enzyme in transplanted rat gliomas were increased, while the volume of gliomas was decreased in hypertention chemotherapy group compared with chemotherapy alone. CONCLUSION: Chemotherapy with angiotensin II-induced hypertension has a enhancing effect on chemotherapy for improving the drug delivery to tumor tissue by a increased tumor blood flow and enhancing activity of interleukin -1β converting enzyme.     

Effect of capsaicin on liver fibrogenesis in rats

AIM：To investigate the effects of capsaicin on rat hepatic stellate cells (HSCs) in vitro and on the liver fibrogenesis in vivo. METHODS：HSCs were cultured with different concentrations of capsaicin. The levels of reactive oxygen species (ROS) were tested with a DCFH-DA kit. The proliferation of HSCs was detected by CCK-8 assay. The expression of α-smooth muscle actin in HSCs was evaluated by Western blotting. The expression of fibrosis-related genes was detected by RT-PCR. The apoptosis of HSCs was measured by flow cytometry. The rat model of liver fibrosis was established by intraperitoneal injection of carbon tetrachloride. Capsaicin at different concentrations was given by gavage. The pathologic changes of the liver sections were observed under microscope with HE staining. Hydroxyproline content in the liver tissues and the levels of collagen Ⅲ and hyaluronic acid in the serum were also measured. RESULTS：Capsaicin inhibited the generation of ROS in a concentration-dependent manner. Compared with control, the proliferation and activation of HSCs were inhibited (P<0.05) and the apoptosis of HSCs was promoted by capsaicin (P<0.05). Capsaicin down-regulated the expression of tissue inhibitor of metalloproteinase 1 and transforming growth factor β 1 in activated HSCs (P<0.05). Capsaicin decreased the levels of hydroxyproline, collagen III and hyaluronic acid in the rats (P<0.05). CONCLUSION：Capsaicin inhibits the proliferation and activation, and promotes the apoptosis of hepatic stellate cells, thus down-regulating the fibrogenesis level of the liver in rats．     

Expression of Cdc25a in cross-species hepatocellular carcinoma tissues and its significance

AIM：To study the expression of cell division cycle 25a protein (Cdc25a) in hepatocellular carcinoma (HCC) tissues of different species, including human, rat and tree shrew, and to verify the feasibility of the strategy of cross-species oncogenomics screening. METHODS：Real-time fluorescence quantitative PCR and Western blotting were applied to detect the expression of Cdc25a at mRNA and protein levels in the HCC tissues, corresponding HCC-adjacent liver tissues and normal liver tissues collected from humans, rats and tree shrews. RESULTS：The mRNA expression of Cdc25a in the HCC tissues of humans, rats and tree shrews was higher than that in normal liver tissues (P<0.05). The mRNA levels of Cdc25a in the HCC tissues of humans and rats were higher than those in the corresponding HCC-adjacent liver tissues (P<0.05). The mRNA expression of Cdc25a in the HCC tissues was significantly correlated with the portal vein tumor thrombus, the extrahepatic metastasis and the clinical stage, but was not correlated with the recurrence of tumor, the diameter of tumor, the number of tumor, the level of serum alpha-fetoprotein and the differentiation of tumor. The protein levels of Cdc25a in the HCC tissues of humans, rats and tree shrews were higher than those in the corresponding HCC-adjacent liver tissues and the normal liver tissues. CONCLUSION：Cdc25a may be a particularly crucial molecule for hepatocarcinogenesis. The cross-species oncogenomics screening may represent a feasible and convenient way for identifying key molecules of human HCC.     

Ferulic acid protects against apoptosis of PC12 cells induced by kainic acid

AIM：To investigate the effect of ferulic acid (FA) on the apoptosis of PC12 cells induced by kainic acid (KA) in vitro. METHODS：In order to establish an Alzheimer disease neuronal cell model, the rat pheochromocytoma cell line PC12 was treated with KA at a concentration of 50 μmol/L. These model neurons were divided into KA model group and 3 groups treated with FA at doses of 25, 50 and 100 μmol/L, respectively. At the same time, normal group was established without KA pretreatment. The viability of the PC12 cells was detected by MTT assay. The expression of Bcl-2, Bax and cytochrome C (Cyt C) was determined by immunocytochemical method. Apoptotic rate of the PC12 cells was measured by flow cytometry with annexin V/PI double staining. The protein levels of Bcl-2, Bax and Cyt C were analyzed by Western blotting. RESULTS：The cell survival rate, the expression of Bcl-2 and the ratio of Bcl-2 to Bax in KA model group were significantly decreased (P<0.01),while the expression of Bax and Cyt C was obviously increased compared with normal control group (P<0.01). The apoptotic rate in KA model group was obviously increased compared with normal control group (P<0.01) After the intervention of FA, the cell survival rates were increased and the apoptotic rates were decreased. Furthermore, the positive rate and expression of Bcl-2, and the ratio of Bcl-2 to Bax in each dose of FA treatment group were significantly increased, while the expression of Bax and Cyt C in each dose group was significantly reduced as compared with KA model group (P<0.05 or P<0.01). CONCLUSION：KA obviously induces apoptosis of PC12 cells. FA had obvious protective effect on PC12 cells against the toxicity of KA. FA blocks endogenous apoptic pathway through inhibiting the expression of Bax and Cyt C and increasing the expression of Bcl-2 and the ratio of Bcl-2/Bax, thus improving the survival rate of PC12 cells.     

Effect of scutellarin on VEGF expression in human retinal pigment epithelial cells and retinas of diabetic rats

AIM: To evaluate the influence of scutellarin on the expression of vascular endothelial growth factor (VEGF) in high glucose-treated human retinal pigment epithelial cell line ARPE-19 and to observe the effects of scutellarin on the protein expression of VEGF, p-ERK and VEGFR2 in the retinas of type II diabetic rats. METHODS: Cultured ARPE-19 cells were divided into normal control group, scutellarin group, high glucose group and high glucose+scutellarin group. The protein levels of VEGF, p-ERK and VEGFR2 were measured by Western blot. The VEGF release in ARPE-19 cells was detected by ELISA. Normal rats were randomly divided into normal control group and scutellarin group. Diabetic rat model was established by feeding with high-fat diet and injecting with streptozocin, and randomly divided into diabetes group and diabetes treated with scutellarin group. After 16 weeks, the eyes were removed. The morphological changes of the retinas were observed under light microscope with HE staining, and histopathological score was recorded. The expression of VEGF in the retinas was observed by the method of immunohistochemistry. RESULTS: Compared with normal control group, the protein levels of VEGF, p-ERK and VEGFR2 in the ARPE-19 cells decreased in scutellarin group, but increased in high glucose group. The histopathological score of the retinas showed significant difference among diabetes group, diabetes treated with scutellarin group and normal control group, and no significant difference between normal control group and scutellarin group was observed. The expression of VEGF increased in diabetic group and was significantly higher than that in scutellarin treatment group (P<0.05). CONCLUSION: Scutellarin inhibits the increased protein le-vels of VEGF, p-ERK and VEGFR2 in ARPE-19 cells, and decreases the expression of VEGF in the retinas of diabetic rats. The suppression of the diabetic retinopathy development by scutellarin may be partly involved in the ERK/MAPK pathway.     

Effect of ghrelin in septal nucleus on gastric motility of diabetic gastroparesis rats and its potential mechanism regulated by hypothalamic arcuate nucleus

AIM：To study the effect of ghrelin in septal nucleus on the gastric motility of the rats with diabetes mellitus (DM) and to investigate the regulation of ghrelin pathway between arcuate and septal nucleus nuclei on gastric motility. METHODS：Streptozotocin was injected intraperitoneally to establish a DM rat model. Fluorescence immunohistochemistry and real-time PCR were used to detect the expression of ghrelin receptor GHS-R1a. The gastric motility was evaluated by implantation of a force transducer on the surface of rats stomachs and the motility index was also calculated. The neural connections between arcuate and septal nuclei were analyzed by the technique of fluorogold tracing. The neural control pathway of gastric motility was determined by central drug injection, nucleus lesion or nucleus electrical stimulation. RESULTS：The expression of GHS-R1a in the septal nucleus of DM rats was lower than that in normal rats (P<0.05). The amplitude and frequency of gastric motility in the DM rats were lower than those in the normal rats (P<0.05). The gastric motility of normal and DM rats were increased by injection of ghrelin into the septal nucleus in a dose-dependent manner. Seven days after injection of fluorogold into the septal nucleus, some neurons in arcuate nucleus were labeled by fluorogold and part of the labeled neurons were ghrelin immunopositive. No effect of nucleus lesion or nucleus electrical stimulation on the gastric motility in the normal rats was observed. In DM rats, the lesion of septal nucleus decreased the gastric motility (P<0.05). In the normal rats, the change of gastric motility caused by electrical stimulation in arcuate nucleus was not affected by the lesion of septal nucleus (P>0.05), while the change was attenuated in DM rats (P<0.05). The ghrelin receptor antagonist ［D-Lys-3］-GHRP-6 had no significant effect on the gastric motility induced by electrical stimulation in arcuate nucleus of the normal rats (P>0.05), but it reduced the change in the DM rats (P<0.05). CONCLUSION:Ghrelin in septal nucleus and the ghrelinergic pathway between arcuate and septal nuclei play an important role in the modulation of gastric motility in DM rats.     

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.     

Attenuated Salmonella carrying pGRIM-19-si-survivin co-expression plasmid inhibits growth of prostate cancer subcutaneous xenografts in vivo

AIM：To explore the effects of pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella on prostate cancer subcutaneous xenograft growth in nude mice. METHODS：Prostate cancer xenograft model was established in nude mice. Co-expression plasmids carried by attenuated Salmonella were introduced by intraperitoneal injection. The xenograft volumes were monitored timely. Immunohistochemical staining, RT-PCR and TUNEL assay were applied to investigate the related mechanisms that pGRIM-19-si-survivin inhibited tumor growth in vivo. RESULTS：Compared with psi-survivin and pGRIM-19 carried by attenuated Salmonella (control groups), the tumor volumes were reduced markedly in pGRIM-19-si-survivin plasmid group. The mean shrinkage rates were 2.36 and 3.02 times. pGRIM-19-si-survivin co-expression plasmid carried by attenuated Salmonella inhibited survivin expression but strengthened GRIM-19 expression obviously (P<0.05). The mRNA expression of apoptosis-related proteins such as Bcl-xL, Stat3, cyclin D1 and c-Myc was inhibited, and the vascular endothelial growth factor (VEGF) mRNA and Ki67 protein were also inhibited, but the caspase-3 mRNA expression was up-regulated (P<0.05) with significant cell apoptosis. CONCLUSION： pGRIM-19-si-survivin co-expression plasmid carried by human attenuated Salmonella inhibits the growth of prostate cancer subcutaneous xenografts in nude mice by promoting cell apoptosis and inhibiting prostatic cancer proliferation.     

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.     

Effects of Shenmai injection on myocardial fibrosis in rat model of diabe-tic cardiomyopathy

AIM：To explore the effects of Shenmai injection on myocardial fibrosis in a rat model of diabetic cardiomyopathy (DCM). METHODS：Wistar rats (n=30) were randomly divided into control group, diabetes group and treatment group. Single intraperitoneal injection of streptozotocin was utilized to establish a rat model of DCM. The rats with DCM in treatment group were intraperitoneally injected with Shenmai injection. Ventricular cannulation was applied to assess the cardiac functions. The formation of collagen in the cardiac tissues was assessed by Masson staining. The generation of reactive oxygen species (ROS) in the cardiac tissues was detected by dihydroethidium staining. The expression levels of matrix metalloproteinase 2 (MMP-2), tissue inhibitor of metalloproteinase 2 (TIMP-2) and collagen I in the cardiac tissues were determined by Western blotting. RESULTS：Compared with control group, the cardiac functions were deteriorated in diabetes group (P<0.05), which was improved in treatment group as compared with diabetes group (P<0.05). Compared with control group, the formation of collagen and ROS increased significantly in diabetes group (P<0.05), which was decreased in treatment group as compared with diabetes group (P<0.05). Compared with control group, the expression level of MMP-2 in the cardiac tissues was deceased and TIMP-2 was increased significantly in diabetes group (P<0.05), but reversed significantly in treatment group (P<0.05).CONCLUSION:Shenmai injection attenuates cardiac fibrosis in the rats with DCM by inhibiting the generation of ROS.     

Expression and significance of TLR4 in Sombatis cell-based model of epilepsy

AIM：To study the change of Toll-like reporter 4 (TLR4) expression in Sombatis cell-based model of epilepsy and to explore the role of TLR4 in this model. METHODS：After cultured in vitro for 9 d, the neurons of newborn SD rats were randomly divided into control group and model group. The neurons in model group were cultured in low-magnesium medium for 3 h and then returned to the normal medium culture. The expression of TLR4 at mRNA and protein levels was detected by fluorescence quantitative PCR and the method of immunohistochemistry, respectively. RESULTS：The immunohistochemical results showed that the protein expression of TLR4 was enhanced in the Sombati’s cells. The results of fluorescence quantitative PCR showed that the mRNA expression of TLR4 time-dependently increased in the Sombatis cells (P<0.05). CONCLUSION：The expression of TLR4 in the neurons is up-regulated in Sombatis cell-based model of epilepsy in rats, suggesting that TLR4 is associated with the pathogenesis of epilepsy.     

Rapamycin markedly slows disease progression in a rat model of passive Heymann nephritis

AIM：To determine the effect of rapamycin on the progression of passive Heymann nephritis (PHN), and whether autophagy is involved in this process. METHODS：Male Sprague-Dawley rats (n=24) were randomly divided into 3 groups: control group, PHN group and rapamycin treatment group. The rat PHN model was induced by injection of anti-Fx1A serum through penile vein, and all rats were sacrificed on day 21. Automatic biochemical analyzer was used to detect 24 h urine protein, blood urea nitrogen and serum creatinine. Renal damage was observed through per-iodic acid-silver methenamine staining. The number of podocyte was estimated by Weibel-Gomez method. The glomerular deposition of C5b-9, the expression of caspase-3 and expression of autophagy marker LC3 in glomeruli were examined by immunofluorescence staining, immunohistochemical staining and Western blotting, respectively. RESULTS：Rapamycin significantly reduced proteinuria in the PHN rats (P<0.05), while the renal functions in 3 groups were normal, without significant difference. Although rapamycin limited weight gain in the rats, the health of the rats during drug treatment was not affected. Rapamycin retarded glomerular basement membrane thickening in the PHN rats. Rapamycin significantly reduced the podocyte deletion by preventing podocyte apoptosis. Rapamycin enhanced the level of autophagy of glomerular inherent cells. CONCLUSION：In the disease process of PHN, appropriate strength of autophagy plays a protective role. Rapamycin appropriately enhances autophagy and prevents podocyte apoptosis, thus reducing nephropathy and proteinuria. This may be one of the important mechanisms of rapamycin to slow down the progress of PHN.