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Objective:To evaluate the role of signal transducer and activator of transcription 3 (STAT3)/nuclear receptor coactivator 4 (NCOA4)-mediated ferritinophagy in salidroside-induced attenuation of intestinal ischemia-reperfusion (I/R) injury in mice and the relationship with ferroptosis.Methods:Thirty-six SPF-grade healthy male C57BL mice, aged 6-8 weeks, weighing 20-25 g, were divided into 6 groups ( n=6 each) by a random number table method: sham operation group (S group), sham operation+ salidroside group (SS group), intestinal I/R group (IR group), intestinal I/R+ salidroside group (IS group), intestinal I/R+ salidroside+ autophagy activator rapamycin group (ISR group) and intestinal I/R+ salidroside+ STAT3 activator colivelin group (ISC group). The intestinal I/R injury model was established by clamping the superior mesenteric artery for 45 min followed by 30-min reperfusion in IR, IS, ISR and ISC groups, while the superior mesenteric artery was only isolated without clipping in S and SS groups. At 1 week before developing the model, salidroside 40 mg/kg was intraperitoneally injected once a day for 7 consecutive days in SS, IS, ISC and ISR groups, rapamycin 4 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISR, colivelin 1 mg/kg was intraperitoneally injected once a day for 7 consecutive days in group ISC, while the equal volume of normal saline was given instead in the rest two groups. The mice were sacrificed at 30 min of reperfusion, and intestinal tissues were obtained for examination of the pathological changes after HE staining (with a optical microscope) which were scored according to Chiu and for determination of contents of malondialdehyde (MDA), Fe 2+, glutathione (GSH) and reactive oxygen species (ROS), activity of superoxide dismutase (SOD) and expression of p-STAT3, STAT3, glutathione peroxidase 4 (GPX4), NCOA4 and ferritin heavy chain 1 (FTH1) in intestinal tissues (by Western blot). Results:Compared with group S, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, the content of GSH was decreased, the activity of SOD was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, the p-STAT3/STAT3 ratio was increased ( P<0.05), pathological injury was found in intestinal tissues, and no significant change was found in the aforementioned indexes in group IR( P>0.05). Compared with group IR, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly decreased, GSH content was increased, SOD activity was increased, the expression of p-STAT3 and NCOA4 was down-regulated, the expression of GPX4 and FTH1 was up-regulated, p-STAT3/STAT3 ratio was decreased ( P<0.05), and the pathological injury was significantly alleviated in intestinal tissues in group IS. Compared with group IS, the Chiu′s score and contents of MDA, Fe 2+ and ROS were significantly increased, GSH content was decreased, SOD activity was decreased, the expression of p-STAT3 and NCOA4 was up-regulated, the expression of GPX4 and FTH1 was down-regulated, p-STAT3/STAT3 ratio was increased ( P<0.05), and the pathological injury was aggravated in intestinal tissues in ISR and ISC groups. There was no statistically significant difference in the expression of STAT3 among the five groups ( P>0.05). Conclusions:STAT3/NCOA4-mediated ferritinophagy is involved in the process of salidroside-induced reduction of intestinal I/R injury in mice, which may be related to inhibiting ferroptosis.
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Objective:To establish a method for quantitative analysis of the active ingredients including salidroside, rosarin and rosavin and content determination in Rhodiola rosea at different harvest months. Methods:HPLC was used on an X selectHSS T3 (250 mm×4.6 mm, 5 μm) column with mobile phase consisting of methol-acetonitrile-phosphoric acid (0.05%) aqueous solution for gradient elution at a flow rate of 1 ml/min. The wavelength was detected at 275 nm (salidroside) and 254 nm (rosarin, rosavin). The column temperature was set at 30 ℃ and the injection volume was 5 μl.Results:The peak areas of Salidroside, rosarin and rosavin showed good linear relationships ( r > 0.999) with the content in the ranges of 44-1 420, 10-307 and 18-573 μg, respectively. The method was precise, stable, repeatable and the sample recovery test all well satisfied the requirements of quantitative analysis. The highest accumulation of the active ingredients was observed in Rhodiola rosea in September and the content of salidroside, rosarin and rosavin were 0.66, 0.07 and 0.53 mg/g, respectively. Conclusion:This method is simple and rapid to evaluate the content of active ingredients in Rhodiola rosea.
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Objective To research the effect and autophagy in hepatic ischemia-reperfusion injury based on relevant indicators of the specimens of rat liver which ischemia reperfusion model by salidroside pretreatment. Methods A total of 90 male SD rats were randomly divided into the sham group, the model group, the low, medium and high dose group, 18 rats in each group. The low, medium and high dose group rats were treated with 7.5, 15, 30 mg/kg salidroside solution by gavage, and the sham group and model group and model group were filled with saline in the same volume,one time per day. After 7 days, all the rats were set up with the model of IR except the rats in sham groups. The AST and ALT of serum, contrast between groups liver tissue by Optical microscope with HE dyeing at 4, 8, 16 h after reperfusion. Western Blot was used to detect the expression of protein of LC3 and Beclin-1. The number and morphology of autophagy in each group of liver cells were observed by electron microscopy. Results After reperfusion 4, 8, 16 h, the level of ALT (662.36 ± 5.82 U/L vs. 983.67 ± 8.96 U/L, 436.49 ± 12.93 U/L vs. 1536 ± 10.77 U/L, 168.61 ± 8.34 U/L vs. 280.42 ± 17.37 U/L) of the high dose group weresignificantly lower than the model group, and the AST (513.29 ± 11.74 U/L vs. 656.38 ± 7.67 U/L, 276.29 ± 9.21 U/L vs. 930.19 ± 15.62 U/L, 97.83 ± 4.29 U/L vs. 211.23 ± 7.87 U/L) of the high dose group were significantly lower than the model group. After reperfusion 8, 16 h, the expression of LC3-Ⅱ (1.21 ± 0.16 vs. 1.91 ± 0.12, 2.00 ± 0.14 vs. 1.09 ± 0.11) in the high dose group were significantly lower than the model group, and the results were same to Beclin1 (3.53 ± 0.19 vs. 7.15 ± 0.14, 2.65 ± 0.27 vs. 7.60 ± 0.21) (P<0.05). After reperfusion 8 h, the number of autophagosome (3.24 ± 0.62 vs.7.84 ± 0.45) in the high dose group were significantly lower than the model group (P<0.05). Conclusions The hepatic ischemia-reperfusion injury was serious, and inhibiting autophagy was one of possible mechanisms to protect liver cells by salidroside.
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Objective To establish a method for the determination of protocatechuic acid, salidroside, and chlorogenic acid in Sargentodoxa cuneata. Methods The separation was performed on a Waters XSELECT CSH C18 (150 mm × 4.6 mm, 5 μm) with methanol-acetonitrile-0.2 % phosphoric acid as the mobile phase in a gradient elution at a flow rate of 0.8 ml/min. The detection wavelength was 260 nm and the column temperature was 35 ℃. Results The linear ranges of protocatechuic acid, salidroside, and chlorogenic acid were 0.0020-0.0120, 0.0600-0.3602, 0.0750-4.5006 mg/ml, respectively. The average recoveries were 98.01% (RSD=0.07%), 98.53 % (RSD=0.12%), and 101.10 % (RSD=1.92%), respectively. Conclusions The method is simple, accurate, and highly reproducible, which could provide the scientific evidence for the quality control of Sargentodoxa cuneata.
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Objective To explore the role of salidroside in HIRI and its related mechanism. MethodsA total of 90 male SD rats were randomly divided into the sham group, the model group, the low, medium and high dose group, 18 rats in each group. The low, medium and high dose group rats were injected with 7.5, 15, 30 mg/kg salidroside solution, and the sham group and model group were injected with saline in the same volume, one time per day. After 7 days, all the rats were set up with the model of IR except the rats in Sham groups. The AST and ALT of serum, contrast between groups liver tissue by Optical microscope with HE dyeing at 4, 8, 16 h after reperfusion. Western Blot was used to detect the expression of protein of MAPK, JNK, ERK and NF-κB.ResultsFour, 8, 16 h after reperfusion, the level of ALT (540.67 ± 15.91 U/L vs.697.67 ± 5.98 U/L, 307.50 ± 12.97 U/L vs.962.50 ± 17.63 U/L, 103.33 ± 3.95 U/L vs.198.17 ± 9.73 U/L) and AST (651.17 ± 7.39 U/L vs.944.67 ± 11.38 U/L, 415.50 ± 10.97 U/L vs.1561.83 ± 15.76 U/L, 168.33 ± 5.81 U/L vs. 280.33 ± 12.35 U/L) in the high dose group were significantly lower than those in the model group. Eight and 16 hours after reperfusion, the expression of MAPK (1.28 ± 0.19 vs. 2.10 ± 0.12, 1.64 ± 0.14 vs.1.89 ± 0.14), JNK (1.80 ± 0.10 vs. 2.42 ± 0.11, 0.84 ± 0.17 vs. 3.32 ± 0.19), ERK (2.43 ± 0.10 vs.5.95 ± 0.09, 2.07 ± 0.13 vs. 6.61 ± 0.14), NF-κB (2.32 ± 0.16 vs. 3.08 ± 0.10, 2.11 ± 0.13 vs. 2.74 ± 0.17) in the high dose group were significantly lower than the model group (P<0.05).Conclusions The salidroside could reduce the liver ischemia- reperfusion injury, and its mechanisms may rugulate the MAPK/NF-κB signaling pathways.
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Objective To investigate the protection of Rhodioloside on brain tissue in diabetic rats. Methods The rats were randomly divided into six groups: a normal control group, a model control group, a Rhodioloside high, medium and low dose group, and a positive control group(n=16). Except normal control group, the diabetic models were set up by intraperitoneal injecting STZ(60 mg/kg). And then, the Rhodioloside high, medium, low dose group were respectively given Rhodioloside solution(100、50、25 mg/kg) by intragastrical administration(ig), the positive control group was given metformin hydrochloride(25 mg/kg) by ig, the normal control group and positive control group were given normal saline(NS). Before the drug was given and 4, 8, 12 weeks after, the level of blood sugar was determined. And 12 weeks later, the body weight was detected; the activity of CPK, LDH and the level of T-AOC in serum were determined; the activity of SOD, GSH-Px, CAT and the content of MDA in brain tissue were determined; and the histopathological changes of the brain tissue was observed by HE staining.Results Compared with the model control group, the level of blood sugar (16.0 ± 0.9 mmol/Lvs. 18.0 ± 1.2 mmol/L) in Rhodioloside high dose group was significantly decreased (P0.05). The brain histopathological changes of Sal treated groups were significantly improved, while the treatment effect of Rhodioloside high dose group was the most significant.Conclusions Rhodioloside had dose-dependent protective effect on the brain tissue in diabetic rats, whose mechanism perhaps related to its effects on enhancing the activity of antioxidant enzymes, and reducing the damage of oxidative stress.
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Objective To investigate the effect of salidroside on brain edema and neurological function in global cerebral ischemia-reperfusion injury in rats.Methods A total of 100 Sprague Dawley rats were randomly divided into sham operation,ischemia-reperfusion and salidroside 12,24 and 48 mg/kg groups (n =20 in each group),and than redivided into 6 h,24 h,72 h and 7 dsubgroups (n =5 in each subgroup).A rat model of global cerebral ischemia was established using the four-vessel occlusion method.Immediately after modeling,all groups were administered intragastrically for 7 days.The brain water content was quantitated by the wet-dry weight method.The neurological evaluation was performed using a neurological deficit score (NDS).Results After modeling both the ischemia-reperfusion group and all the salidroside groups had significant neurological deficit,and as time went by,it was improved gradually.Compared to the ischemia-reperfusion group at the corresponding time points,neurological deficit in all the salidroside groups was improved significantly (all P ＜ 0.05),and showing a dose-dependent trend.Compared to the salidroside 12 mg/kg and 24 mg/kg groups,neurological deficit in the salidroside 48 mg/kg group was improved significantly at 72 hours and 7 days (all P ＜ 0.05).The brain water contents began to increase at 6 hours after modeling in the the ischemia-reperfusion group and all the salidroside group.They reached the peak at 72 hours,and significantly higher than that in the sham operation group (all P ＜ 0.05).The brain water contents in all the salidroside group were significantly lower than those in the ischemiareperfusion group at 24 and 72 hours after modeling (all P ＜ 0.05) and showing a dosedependent trend.The brain water content in the salidroside 48 mg/kg group was close to that in the sham operation group at 7 days after modeling.Conclusions Salidroside may significantlydecrease brain edema and improve neurological function in global cerebral ischemia-reperfusion injury in rats,and it has a neuroprotective effect.