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OBJECTIVE：To study the mechanism of improvement effects of Fupi rougan granule （FRG）on hepatic fibrosis model rats. METHODS ：The rats were randomly divided into blank group ，model group ，Colchicine tablet group （chemical positive control ，0.2 mg/kg），Fuzheng huayu capsule group （TCM positive control ，0.415 g/kg），FRG low-dose ，medium-dose and high-dose groups （20，40，80 g/kg），with 10 rats in each group ，except for 11 rats in blank group and model group （one rat was used to judge whether the modeling was successful ）. Except for blank group ，other groups were given intraperitoneal injection of 50％ CCl4 olive oil solution and intragastric administration of 30％ ethanol to induce hepatic fibrosis model. After modeling ， administration groups were given relevant medicine intragastrically ；blank group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. After last administration ，morphology changes of liver tissue in rats were observed. The serum levels of HA ，LN，PCⅢ and Col Ⅳ in rats were detected ，and protein expression of Beclin- 1 and LC3-Ⅱin liver tissue were also determined. mRNA and protein expression of Akt ，AMPK，mTOR，p70S6K were detected in liver tissues of rats. RESULTS ：Compared with blank group ，the structure of hepatic lobules in the model group was disordered ，the proliferation of fibrous tissue was obvious ，and some pseudolobules were formed ；the serum levels of HA ，LN，PCⅢ and Col Ⅳ， the protein expression of Beclin- 1 and LC 3-Ⅱ in liver tissue as well as mRNA and protein expression of Akt ，AMPK，mTOR and p70S6K were increased significantly （P＜0.01）. Compared with model group ，the liver injury of rats in FRG groups was significantly relieved ，and the levels of the above indexes in serum and liver tissue （except for LN and PC Ⅲ in FRG low-dose group） were significantly reduced （P＜0.05 or P＜0.01）. CONCLUSIONS ：FRG can improve hepatic fibrosis in rats ，the mechanism of which may be associated with down-regulating the expression of autophagy associated protein and Akt/AMPK/mTOR/ p70S6K signaling pathway related protein.
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AIM: To analyze the effects of salvianolic acid on autophagy and apoptosis of skeletal muscle ischemia-reperfusion injury mice through AKT/mTOR/p70S6K signaling pathway. METHODS: A total of 45 SPF male rats were randomly divided into sham operation group, model group, salvianolic acid group. Model group and salvianolic acid group were prepared for skeletal muscle ischemia-reperfusion injury model. At 72 h, 48 h, 24 h and 15 min before reperfusion, 1 mL of salvianolic acid solution was administered intraperitoneally at a dose of 30 mg/kg. Rats in the sham operation group and the model group were intraperitoneally injected with 1 mL of normal saline. The pathological morphology of the gastrocnemius muscle, the contents of serum CK and LDH, the expression of Akt, p-Akt, p70S6K, p-p70S6K, mTOR and p-mTOR in the gastrocnemius muscle and the expression of Bcl-2 and Bax in the gastrocnemius muscle were observed. RESULTS: At the 0 h, 4 h and 24 h within reperfusion, the W/D ratio of gastrocnemius muscle in the model group were higher than those in the sham operation group. The W/D ratio of the gastrocnemius muscle in the salvianolic acid group was lower than that in the model group (P<0.05). The levels of serum LDH and CK in the model group were higher than those in the sham operation group at 0 h, 4 h and 24 h after reperfusion. The serum levels of LDH and CK in the salvianolic acid group were lower than those in the model group (P<0.05). The expression of Akt protein in the gastrocnemius of the model group was lower than that in the sham operation group, and the expression of p-Akt protein was higher than that in the sham operation group. The protein expressions of Akt, p-Akt, p70S6K, p-p70S6K, mTOR and p-mTOR in the salvianolic acid group were significantly higher than those of the model group (P<0.05). The IOD value quantified the expression of Bax and Bcl-2 protein in the gastrocnemius muscle cells. The Bcl-2 IOD value and Bcl-2/Bax ratio in the model group were lower than those in the sham operation, whlie the Bax IOD value in the model group was higher than that in the sham operation group. The Bcl-2 IOD value and Bcl-2/Bax ratio in the salvianolic acid group were higher than those in the model group, and the Bax IOD value was lower than that in the model group, the difference was statistically significant (P<0.05). CONCLUSION: Salvianolic acid can maintain the homeostasis of Bax and Bcl-2 in bone cells of rats with skeletal muscle ischemia-reperfusion injury and inhibit cell apoptosis. The mechanism may be related to the activation of AKT/mTOR/p70S6K signaling pathway.
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Objective To observe the effect of Acanthus ilicifolius alkaloid A (HBOA) on PI3K/AKT/mTOR/p70S6K signaling pathway in rats with hepatic fibrosis induced by carbon tetrachloride (CCl4), and to explore the mechanism of action of HBOA against liver fibrosis. Methods Rats were randomly divided into normal group, model group, high, medium; and low-dose HBOA groups (100, 50, 25 mg/kg), and colchicine group (0.4 mg/kg). Except for the normal group, the rats in other groups were given with a 50% CCl4 olive oil solution twice a week for 12 weeks to induce a rat model of liver fibrosis. From the ninth week of modeling, the drug-administered group was given the corresponding test drug once daily for 4 weeks. After the experiment, the body mass change and liver index were calculated. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the liver homogenate of each group were detected. The protein expression of p-PI3K, p-Akt, p-mTOR, and p-p70S6K in liver tissue was detected by Western blotting. Results Compared with the model group, the body weight of mice of each drug-administered group was significantly increased, and the liver index, and ALT and AST levels were decreased in liver tissue. In addition, HBOA high and medium-dose groups significantly inhibited the protein expression of p-PI3K, p-Akt, p-mTOR, and p-p70S6K. Conclusion HBOA has a protective effect on hepatic fibrosis rats, and its mechanism may be related to the inhibition of PI3K/Akt/mTOR/p70S6K signaling pathway.
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Chronic cerebral hypoperfusion (CCH), which is associated with onset of vascular dementia, causes cognitive impairment and neuropathological alterations in the brain. In the present study, we examined the neuroprotective effect of duloxetine (DXT), a potent and balanced serotonin/norepinephrine reuptake inhibitor, on CCH-induced neuronal damage in the hippocampal CA1 region using a rat model of permanent bilateral common carotid arteries occlusion. We found that treatment with 20 mg/kg DXT could attenuate the neuronal damage, the reduction of phosphorylations of mTOR and p70S6K as well as the elevations of TNF-α and IL-1β levels in the hippocampal CA1 region at 28 days following CCH. These results indicate that DXT displays the neuroprotective effect against CCH-induced hippocampal neuronal death, and that neuroprotective effect of DXT may be closely related with the attenuations of CCH-induced decrease of mTOR/p70S6K signaling pathway as well as CCH-induced neuroinflammatory process.
Subject(s)
Brain , CA1 Region, Hippocampal , Carotid Artery, Common , Cognition Disorders , Dementia, Vascular , Duloxetine Hydrochloride , Models, Animal , Neurons , Neuroprotection , Neuroprotective Agents , Phosphorylation , Ribosomal Protein S6 Kinases, 70-kDaABSTRACT
Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.