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Aim: To explore the mechanism of icariside II (ICS II) on improving left ventricular function based on endoplasmic reticulum stress and caspase-12 signaling in spontaneously hypertensive rats (SHRs). Methods: Thirty 14-week-old male SHRs were divided into model group, ICS II low, middle and high dose groups and positive drug group (n = 6). WKY was used as control group (n =6). ICS II groups were respectively given ICS 114, 8, 16 mg · kg-1(ig, qd), and positive drug group was given losartan (20 mg · kg-1). At the end of 26th weeks, anesthetized rats were measured by ultrasound for detection of the left ventricular function, RT-PCR was used to determine the level of GRP78 mR- NA in the left ventricle tissue, and Western blot was used to assess the levels of GRP78 and cleaved-caspase- 12/9/3 protein in the left ventricle tissues. Results: Compared with WKY group, the internal diameter and posterior wall thickness of the left ventricular end diastolic increased, while the ejection fraction and fractional shortening decreased in SHR group. GRP78 mRNA and protein levels were up-regulated, and the levels of cleaved-caspase-12/9/3 protein were raised in left ventricle (P < 0.05). Compared with SHR group, the internal diameter and posterior wall thickness of the left ventricular end diastolic increased in ICS II medium and high dose groups and positive drug group (P <0. 05), while the ejection fraction and fractional shortening decreased (P<0.05). GRP78 mRNA and protein levels were down-regulated, the levels of cleaved-caspase-12/ 9/3 protein declined in left ventricle (P <0.05). Conclusions: ICS II could improve left ventricular function in SHRs, and its mechanism may be related to improving left ventricular endoplasmic reticulum stress and down-regulating the elevated caspase-12 signaling.
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Aim To investigate the effects of IcarisideⅡ ( ICS Ⅱ) on myocardial fibrosis in spontaneously hypertensive rat( SHR) . Methods Twenty male SHR rats were randomly divided into the model group (group SHR) , ICS Ⅱ low ( ICS Ⅱ-L) , middle ( ICSⅡ-M) and high ( ICS Ⅱ-H) group, and male WKY rats were set as control group ( group WKY) . ICS Ⅱ-L, ICSⅡ-M and ICSⅡ-H groups were intragastrically administered with ICS Ⅱ for 12 weeks. After that the blood pressure was measured in rats. Then, the rats were sacrificed and the left ventricles were separated in order to calculate the left ventricular mass index. Mas-son staining was used to detect the occurrence of inter-stitial fibrosis in cardiac tissues. Real time PCR was used to observe the gene expression of MMP-2, MMP-9 and TIMP-1 in the left ventricle in SHRs. The protein expression levels of MMP-2, MMP-9, TIMP-1, Colla- gen Ⅰ and Collagen Ⅲ were measured by Western blot. Results Compared with SHR group, the myo-cardial fibrosis was reduced after ICS Ⅱ (8, 16 mg· kg-1) treatment. The blood pressure and left ventricu-lar mass index decreased(P<0.05). The expressions of MMP-2, MMP-9, CollagenⅠand CollagenⅢwere down-regulated in left ventricular tissues( P <0.05 ) , while the expression of TIMP-1 was up-regulated( P<0.05) . Conclusion Icariside Ⅱ ameliorates myocar-dial fibrosis in SHR, and the mechanisms might be re-lated to the decrease of blood pressure and down-regu-lation of MMP-2, MMP-9 expression and up-regulation of TIMP-1 expression.
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AIM:To investigate the effects of kaempferol-3-O-rutinoside(KR)on the proliferation,migration of vascular smooth muscle cells(VSMC)and the activation of transforming growth factor βreceptor 1(TGFBR1)signaling pathway in the cells.METHODS: The viability of VSMC was detected by MTT assay.The proliferation of VSMC was measured by EdU staining.The migration ability of VSMC was examined by Transwell assay.The protein levels of the mi-gration-associated proteins matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)were detected by Western blot.Molecular docking study was conducted to explore the interaction between KR and TGFBR 1.The protein le-vels of the phosphorylated TGFBR1,Smad2 and Smad3 were determined by Western blot.RESULTS: KR inhibited the viability of VSMC in a dose-and time-dependent manner.KR reduced the ratio of EdU-positive cells in a dose-dependent manner.KR dose-dependently suppressed the migration ability of VSMC and decreased the protein levels of MMP 2 and MMP9(P<0.05).KR docked into TGFBR1 with the binding energy of -9.804 kcal/mol by forming hydrogen bonds with SER-280,ARG-215,ASP-290 and LYS-335 of TGBFR1.KR dose-dependently suppressed the activation of TGFBR 1 and its downstream proteins Smad2 and Smad3(P<0.05).CONCLUSION: KR inhibits the proliferation and migration of VSMC,possibly via blocking the TGFBR1 signaling pathway.
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Objective To compare the different expressions of miRNAs of alveolar macrophages (AM) in two chronic obstructive pulmonary disease (COPD) models of rat induced by smudging alone or combined with lipopolysaccharide (LPS) infusion. Methods Sixty female Wistar rats were randomly assigned into 4 groups: control group I, control group II, COPD model group I (cigarette smoke exposure alone, CS) and COPD model group II (cigarette smoke exposure + LPS infusion, CS+LPS). COPD rat models were evaluated by chest CT, lung function test and histopathological examination of lungs. The primary AM were acquired and the RNAs were then extracted after carrying out bronchoalveolar lavage. Three pairs of samples were used for detection of miRNAs expression by the method of miRNA microarray chip. The difference was verified by qRT-PCR analysis on another 5 pairs of samples. Data analysis was performed to find out the significantly differential miRNAs expression profiles in COPD rat models. Results The chest CT, lung function test and histopathological examination verified the COPD in rats of CS and CS+LPS groups. Compared with control group I, the expressions of let-7b-3p, miR-376c-3p and miR-675-5p were downregulated in CS group with no miRNAs up-regulated. Compared with control group I, the expressions of let-7b-3p and miR-675-5p were down-regulated, while the expressions of 11 miRNAs were obviously up-regulated in CS+LPS group as miR-200b-3p, miR-665, miR-344b-1-3p, miR-34c-5p, miR-34b-5p, miR-99b-5p, miR-129-1-3p, miR-3557-5p, miR-331-5p, miR-493-5p and miR-200a-3p. Conclusions COPD rat models are established successfully both with CS and CS+LPS. The results of chest CT, lung function test and histopathological examination have shown no significant difference between the two approaches. However, the expressions of miRNAs of AM are significantly different.