Xueshuantong for Injection (Lyophilized, ) Alleviates Streptozotocin-Induced Diabetic Retinopathy in Rats / 中国结合医学杂志

OBJECTIVE@#To investigate the ameliorate effect and underlying mechanism of Xueshuantong for Injection (Lyophilized, , XST) in streptozocin (STZ)-induced diabetic retinopathy (DR) rats.@*METHODS@#Diabetes mellitus (DM) model was induced by intraperitoneal (i.p.) injection of STZ (60 mg/kg) in Sprague-Dawley rats. Diabetic rats were randomized into 3 groups (n=10) according to a random number table, including DM, XST50 and XST100 groups. XST treatment groups were daily i.p. injected with 50 or 100 mg/kg XST for 60 days, respectively. The control and DM groups were given i.p. injection with saline. Blood glucose level and body weight were recorded every week. Histological changes in the retina tissues were observed with hematoxylin-eosin staining. Apoptosis and inflammation related factors, including cleaved caspase-3, glial fifibrillary acidic protein (GFAP), tumor necrosis factor-α (TNF-α) and intercellular cell adhesion molecule-1 (ICAM-1) were detected by Western blot or real-time polymerase chain reaction. Then, the levels of advanced glycation end product (AGE) and its receptor (RAGE) were investigated. Tight junctions proteins (Zonula occludens-1 (ZO-1), Occludin and Claudin-5) of blood-retinal barrier were detected by Western blot. The levels of retinal fifibrosis, transforming growth factor-β1 (TGF-β1)-Smad2/3 signaling pathway were evaluated at last.@*RESULTS@#There was no signifificant difference in the body weight and blood glucose level between XST and DM groups (P>0.05). Compared with the DM group, XST treatment signifificantly increased the retinal thickness of rats (P<0.05 or P<0.01), and suppressed cleaved caspase-3 expression (P<0.01). XST increased the protein expressions of ZO-1, Occludin and Claudin-5 and decreased the mRNA expressions of matrix metalloproteinase 2 (MMP-2) and MMP-9 (P<0.05 or P<0.01). Moreover, XST signifificantly reduced the productions of AGE and RAGE proteins in the retina of rats (P<0.05 or P<0.01), suppressed the over-expression of TNF-α, and decreased the elevated level of ICAM-1 in retina of rats (P<0.05 or P<0.01). XST signifificantly reduced the levels of α-smooth muscle actin (α-SMA), connective tissue growth factor (CTGF), TGF-β1 and phosphorylation of Smad2/3 protein in rats (P<0.05 or P<0.01).@*CONCLUSIONS@#XST had protective effects on DR with possible mechanisms of inhibiting the inflammation and apoptosis, up-regulating the expression of tight junction proteins, suppressing the productions of AGE and RAGE proteins, and blocking the TGF-β/Smad2/3 signaling pathway. XST treatment might play a role for the future therapeutic strategy against DR.

Effect of ligustrazine nanoparticles nano spray on transforming growth factor-β/Smad signal pathway of rat peritoneal mesothelial cells induced by tumor necrosis factor-α / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of ligustrazine nanoparticles nano spray (LNNS) on transforming growth factor β (TGF-β)/Smad signal protein of rat peritoneal mesothelial cells (RPMC) induced by tumor necrosis factor α (TNF-α), and the anti-adhesion mechanism of LNNS in the abdominal cavity.</p><p><b>METHODS</b>The primary culture and subculture of rat peritoneal mesothelial cells (RPMC) was processed by trypsin digestion method in vitro. The third generation was identifified for experiment and divided into 5 groups: a blank group: RPMC without treatment; a control group: RPMC stimulated with TNF-α; RPMC treated by a low-dosage LNNS group (2.5 mg/L); RPMC treated by a medium-dosage LNNS group (5 mg/L); and RPMC treated by a high-dosage LNNS group (10 mg/L). Reverse transcription-polymerase chain reaction was applied to test the expression of fifibronectin, collagen I (COL-I), TGF-β mRNA, and Western blot method to test the Smad protein 7 expression of RPMC.</p><p><b>RESULTS</b>Compared with the blank group, a signifificant elevation in fifibronectin (FN), COL-I and TGF-β mRNA expression of RPMC were observed in the control group (P<0.05). Compared with the control group, LNNS suppressed the expressions of FN, COL-I and TGF-β mRNA in a concentrationdependent manner (P<0.05). The expression of Smad7 protein of RPMC was down-regulated by TNF-α stimulation, and up-regulated with the increase of LNNS dose (P<0.05).</p><p><b>CONCLUSIONS</b>TNF-α may induce changes in RPMC's viability, leading to peritoneal injury. LNNS could reverse the induction of fifibrosis related cytokine FN, COL-I and TGF-β, up-regulating the expression of Smad7 by TNF-α in RPMC, thus attenuate peritoneal injury by repairing mesothelial cells.</p>

Heparin attenuates lipopolysaccharide-induced acute lung injury by inhibiting nitric oxide synthase and transforming growth factor-β/Smad signaling pathway / 中华危重病急救医学

Objective To investigate whether heparin has a beneficial effect on lipopolysaccharide(LPS)-induced acute lung injury(ALI)in rats,and to explore the possible underlying mechanisms. Methods Thirty-two adult Sprague-Dawley(SD)rats were randomly assigned into the control,heparin control,model,and heparin treatment groups,with 8 in each group. ALI rat model was reproduced by intratracheal instillation of LPS at a dose of 1 mg/kg. The rats in the control and heparin control groups received an equal volume of normal saline at the same times. The rats in the heparin control and heparin treatment groups were intravenously received 50 U/kg heparin every 1 hour after the induction of ALI. Animals were sacrificed 24 hours after LPS challenge. Bronchoalveolar lavage fluid(BALF) and lung tissue samples were collected. Histopathological evaluation,lung wet/dry(W/D)ratio,malondialdehyde (MDA),nitric oxide(NO)and myeloperoxidase(MPO)were analyzed. Enzyme-linked immunosorbent assay(ELISA) was used to measure the concentration of inflammatory factor in BALF. Expression of inducible nitric oxide synthase (iNOS)mRNA in the lung of rats was measured by reverse transcription-polymerase chain reaction(RT-PCR). Western Blot was used to determine the expression of transforming growth factor-β1(TGF-β1)and phosphorylation of Smad in the lung tissues. The expression of iNOS in lung was determined by immunohistochemistry. Results In the control and heparin control groups,lung tissue showed a normal structure and clear pulmonary alveoli under a light microscope. In the model group,ALI characters such as extensive thickening of the alveolar wall,significant infiltration of inflammatory cells,demolished structure of pulmonary alveoli,and hemorrhage were found. In the heparin treatment group,heparin treatment markedly alleviated LPS-induced these pathological changes in lung. Compared with control and heparin control groups,lung W/D ratio,lung MDA,NO and MPO levels,and tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6)in BALF in the model group were increased significantly. Compared with the model group, lung W/D ratio,lung MDA,NO and MPO levels,and TNF-αand IL-6 in BALF in the heparin treatment group were significantly decreased〔W/D ratio:7.54±0.17 vs. 10.69±0.15,MDA(mmol/mg):2.01±0.30 vs. 2.51±0.25,NO (μmol/L):3.07±0.21 vs. 3.89±0.14,MPO(U/g):1.94±0.09 vs. 2.74±0.20,TNF-α(μg/L):201.80±0.27 vs. 297.53±0.34,IL-6(μg/L):38.41±0.25 vs. 46.31±0.31,all P<0.05〕. RT-PCR showed that the expression of iNOS mRNA in the heparin treatment group was significantly lower than that in the model group(2-ΔΔCt:3.04±0.18 vs. 4.37±0.15,P<0.05). Western Blot showed that compared with control group,the protein expressions of iNOS and TGF-β1,and phosphorylation of Smad2 and Smad3 were significantly increased,and the heparin could inhibit the protein expressions compared with model group. Immunohistochemistry showed that positive expressions of iNOS in alveolar epithelial cell and capillary endothelial cell in the heparin treatment group were significantly lower than those in the model group. Conclusion Heparin significantly ameliorated the lung injury induced by LPS in rats via the inhibition of nitric oxide synthase expression and the TGF-β/Smad pathway.