Intervention effects of Jinkui shenqi pills on renal fibrosis model rats and its mechanism / 中国药房

OBJECTIVE To investigate th e intervention effect of Jinkui shenqi pills on renal fibrosis （RF）model rats and its mechanism based on transforming growth factor β1/Smads（TGF-β1/Smads）and TGF-β1/extracellular signal regulated kinase （ERK） signaling pathway. METHODS Male SD rats were given adenine suspension （250 mg/kg）to induce RF model. After modeling ， they were randomly divided into model group ，Colchicine tablet group （positive control ，0.45 mg/kg）and Jinkui shenqi pills low-dose，medium-dose and high-dose groups （0.5，1，2 g/kg），with 10 rats in each group. Other 10 healthy rats were selected as normal group. The rats in administration groups were given the corresponding drugs intragastrically ；normal group and model group were given 0.1％ sodium carboxymethyl cellulose solution ，once a day ，for consecutive 30 d. After last medication ，the serum levels of creatinine （Cr）and blood urea nitrogen （BUN），renal weight and body weight were detected. The ratio of BUN/Cr and renal coefficient were calculated. The pathological morphology of renal tissue in rats were observed. The protein and mRNA expressions of TGF-β1，Smad2，Smad3，ERK1 and ERK 2 were detected. RESULTS Compared with normal group ，serum levels of Cr and BUN and renal coefficient were all increased significantly in model group （P＜0.05），while the ratio of BUN/Cr was decreased significantly （P＜0.05）. The volume of the kidney was significantly increased ，and the surface was seriously granulated. Mesangial hyperplasia ，dilation or atrophy of renal tubules ，accompanied by large-area collagen deposition，could be found. Protein and mRNA expressions of TGF-β 1，Smad2，Smad3，ERK1 and ERK 2 were increased significantly in renal tissue （P＜0.05）. Compared with model group ，above indexes of Jinkui shenqi pills groups were all reversed significantly （P＜0.05）；dilation or atrophy of renal tubules was relieved ，and collagen deposition was reduced to different extents. CONCLUSIONS Jinkui shenqi pills can improve renal function of RF model rats ，the mechanism of which may be associated with inhibiting TGF-β1/Smads and TGF-β1/ERK signaling pathway.

Rutin treats myocardial damage caused by pirarubicin via regulating miR-22-5p-regulated RAP1/ERK signaling pathway.

Our experiments have previously demonstrated that rutin (RUT) can improve myocardial damage caused by pirarubicin (THP). However, the underlying molecular mechanisms remain uncertain. In this study, we developed an microRNA (miRNA) chip by replicating the rat model of THP-induced myocardial injury and identified miR-22-5p and the RAP1-member of RAS oncogene family/extracellular regulated protein kinases (RAP1/ERK) signaling pathway as an object of study. Also, in vivo experiments demonstrated that THP caused abnormal changes in the electrocardiogram, cardiac function, and histomorphology in rats (P < .01). THP also reduces the expression of miR-22-5p (P < .01) and increases the levels of RAP1/ERK signaling pathway-related proteins (P < .01, P < .05). RUT significantly improved THP-induced myocardial damage (P < .01), increased the expression of miR-22-5p (P < .01), and decreased the levels of RAP1/ERK signaling pathway-related proteins (P < .01, P < .05). In vitro studies confirmed that Rap1a is one of the target genes of miR-22-5p. miR-22-5p overexpression in cardiomyocytes can affect the RAP1/ERK pathway and reduce reactive oxygen species production and cardiomyocyte apoptosis caused by THP (P < .01), which is consistent with the effect of RUT. Our results indicate that RUT treats THP-induced myocardial damage, which may be achieved by upregulating miR-22-5p, causing changes in its target gene Rap1a and the RAP1/ERK pathway.

Silencing of PTEN inhibits the oxidative stress damage and hippocampal cell apoptosis induced by Sevoflurane through activating MEK1/ERK signaling pathway in infant rats.

Phosphatase and tensin homolog (PTEN) is a suppressive player in tumor but its concrete role in oxidative stress (OS) damage and cell apoptosis remains much exploration. Thus, this study is conducted to explore the participation of PTEN and its mechanisms in OS damage and cell apoptosis in hippocampal cells.Infant rats were grouped into normal, Sevo, Sevo + si-negative control (NC), Sevo + si-PTEN and Sevo + si-PTEN + PD (MEK1/ERK signaling pathway inhibitor) groups. Infant hippocampal cells were grouped into blank, Sevo, Sevo + si-NC, Sevo + si-PTEN and Sevo + si-PTEN + PD groups. The expressions of PTEN and MEK1/ERK signaling pathway-related proteins were determined. OS-related indices in hippocampal tissues and cells were detected. Cell apoptosis was detected by flow cytometry.Sevoflurane up-regulated PTEN expression and silencing of PTEN activates MEK1/ERK signaling pathway in hippocampal tissues and cells of infant rats. Silencing of PTEN alleviated hippocampal tissue pathological status and inhibited sevoflurane-induced cell apoptosis in hippocampal tissues of infant rats. Silencing of PTEN alleviated OS damage in hippocampal tissues of infant rats. Silencing of PTEN inhibited sevoflurane-induced apoptosis after OS damage in hippocampal cells of infant rats. Silencing of PTEN reduced sevoflurane-induced OS damage in hippocampal cells of infant rats.Our study demonstrates that PTEN silencing inhibits the OS damage and cell apoptosis in hippocampal cells induced by Sevoflurane through activating MEK1/ERK signaling pathway in infant rats.

microRNA-329 reduces bone cancer pain through the LPAR1-dependent LPAR1/ERK signal transduction pathway in mice.

BACKGROUND: Bone cancer pain (BCP) is a common symptom occurring among patients with cancer and has a detrimental effect on their quality of life. Growing evidence has implicated microRNA-329 (miR-329) in the progression of bone diseases. In the present study, we aimed to elucidate the potential effects of miR-329 on BCP in a BCP mouse model via binding to lysophosphatidic acid receptor 1 (LPAR1) through the LPAR1/extracellular signal-regulated kinase (ERK) signaling pathway. METHODS: Initially, a BCP mouse model was established via injection of 4 × 104 murine breast tumor (4T1 cell) cells (4 µl). The interaction between miR-329 and LPAR1 was identified using a bioinformatics website and dual luciferase reporter gene assay. The modeled mice were subsequently treated with miR-329 mimic, LPAR1 shRNA, or both, in order to examine the effect of miR-329 on the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) of mice, the expression of LPAR1/ERK signaling pathway-related genes. RESULTS: The positive expression rate of LPAR1 protein and extent of ERK1/2 phosphorylation were increased in BCP mouse models. LPAR1 is a target gene of miR-329, which can inhibit the expression of LPAR1. In response to miR-329 overexpression and LPAR1 silencing, BCP mice showed increased PWT and PWL, along with decreased LPAR1 expression and ratio of p-ERK/ERK. CONCLUSIONS: Altogether, the results obtained indicated that miR-329 can potentially alleviate BCP in mice via the inhibition of LPAR1 and blockade of the LPAR1/ERK signaling pathway, highlighting that upregulation of miR-329 could serve as a therapeutic target for BCP treatment.

Ipilimumab affectsTlymphocytesandBcl-2mRNAexpression in xenograft tissues of lung cancer-bearing mice by inhibiting TGF-β1/ERK signaling pathway / 中国肿瘤生物治疗杂志

@# Objective: To study the effect of ipilimumab on T lymphocytes and Bcl-2 mRNA expression in lung cancer-bearing mice by inhibiting TGF-β1/ERK signaling pathway. Methods: Forty-five C57 mices inoculated with Lewis lung cancer cells were randomly divided into control group, low dose ipilimumab group and high dose ipilimumab group with 15 mice in each. The low and high dose groups were given 3 mg/kg and 5 mg/kg ipilimumab respectively, while the control group was given 0.9% sodium chloride solution with the same volume. The effects of ipilimumab on TGF-β1/ERK signaling pathway, Bcl-2 mRNA expression, immune function improvement and tumor inhibition in three groups were detected by WB and qPCR. Results: After administration of ipilimumab, the tumor weight and volume of mice in low-dose and high-dose groups were significantly lower than that of the control group, and the tumor inhibition rate increased in a dose-dependent manner (P<0.05). The thymus index and spleen index of mice were significantly higher than that of control group, which also increased in a dose-dependent manner (P<0.05). The levels of CD3+, CD4+, CD4+/CD8+ cells in the high and low dose groups were significantly higher than those in the control group, with significantly higher levels in high dose group compared with the low dose group (P<0.05). The levels of serum inflammatory factors were significantly lower than those in control group, and the levels of serum TNF-α, IL-6 and IL-3 in the high dose group were significantly lower than those in the low dose group (P<0.05). The expressions of TGF-β1, ERK1/2, p-ERK1/2 and MEK in tumor tissues of both high and low dose groups significantly decreased, with more lower levels in high dose group than in low dose groups (all P<0.05), and the positive rate of TGF-β1 expression in high dose group was the lowest. The mRNAexpression of Bcl-2 in tumor tissues of high and low dose groups decreased significantly after drug administration, with a significantly lower level in high does group than that in low dose group (P<0.05). Conclusion: Ipilimumab can effectively inhibit TGF-β1/ERK signaling pathway, improve immune function and down-regulate the expression of Bcl-2, thus inhibit the growth of Lewis lung cancer cells and play an antitumor role in mice.

MicroRNA-508 suppresses epithelial-mesenchymal transition, migration, and invasion of ovarian cancer cells through the MAPK1/ERK signaling pathway.

Ovarian cancer (OC) is the sixth most common cancer in women worldwide. Despite advances in detection and therapies, it still represents the most lethal gynecologic malignancy in the industrialized countries. Unfortunately, the molecular events that lead to the development of this highly aggressive disease remain largely unknown. The study explored the ability of microRNA-508 (miR-508) to influence proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) in OC cells. We quantified the level of miR-508 cancer tissues with corresponding adjacent normal tissues collected from 84 patients with OC. Human OC cells SKOV3 and A2780 were treated with negative control (NC), miR-508 mimics, miR-508 inhibitors, and miR-508 inhibitors + a specific MAPK/ERK kinase inhibitor (PD98059) to validate the interaction between miR-508 and MAPK/ERK signaling. The miR-508 expression level was lower while MAPK1 and ERK expression levels were higher in the cancer tissues than in the adjacent normal tissues. Dual-luciferase reporter assay indicated MAPK1 as a target gene of miR-508. The miR-508 mimics reduced the expression of MAPK1, p-MAPK1, ERK, p-ERK and Vimentin, inhibited cell proliferation, migration and invasion, and increased the expression of E-cadherin, while the miR-508 inhibitors resulted in an opposed trend in OC cells. The effects of miR-508 inhibitors on OC cells were lost when the MAPK1/ERK signaling pathway was inhibited by PD98059. Collectively, our data indicate that miR-508 plays a tumor suppressor role in the development and progression of OC and may be a novel therapeutic target against OC.