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Objective: To investigate the anti-inflammatory effects of the total flavonoids from Saussurea involucrata on lipopolysaccharides (LPS)-stimulated murine RAW264.7 macrophages and explore its underlying mechanism of action. Methods: Total flavonoids from Saussurea involucrata were extracted using chromatographic column method. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. The production of nitric oxide was detected by Griess assay and the release of cytokines (IL-10 and TNF-α) and chemokines (MCP-1, MIP-1a, and CCL5/RANTES) was determined by ELISA to evaluate the anti-inflammatory activity of total flavonoids from Saussurea involucrata. Moreover, nuclear translocation of p65, c-Jun, and IRF3 was detected by immunofluorescence microscopy and Western blotting analysis was performed to determine the expression of related proteins. Results: Total flavonoids extracted from Saussurea involucrata were 751.5 mg/g and the content of rutin was 506.5 mg/g. The production of inflammatory mediators including nitric oxide, cytokines, and chemokines was effectively inhibited by total flavonoids from Saussurea involucrata. Meanwhile, total flavonoids also suppressed the nuclear translocation of p65, c-Jun, and IRF3 in LPS-stimulated RAW264.7 cells. The LPS-induced expression of iNOS and COX-2 was remarkably reduced by treatment with total flavonoids from Saussurea involucrata. Moreover, total flavonoids decreased the expression levels of p-IKKa/β, p-TBK1, p-p38, p-ERK, p-JNK, p-p65, p-c-Jun, and p-IRF3 in LPS-exposed RAW264.7 macrophages. Conclusions: Total flavonoids from Saussurea involucrata potentially inhibit the secretion of pro-inflammatory mediators, which may be related to inhibition of p65, c-Jun, and IRF3 signaling pathways in LPS-stimulated RAW264.7 cells.
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OBJECTIVE:To detect the contents of chlorogenic acid and rutoside in Saussurea involucrate,and to optimize the decoction and extraction technology of S. involucrate from different producing areas. METHODS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate from different producing areas were determined by HPLC. L9(34)orthogonal experiment was used to optimize the water amount,decoction times and decoction time using comprehensive score of extraction transport rate of chlorogenic acid and rutoside as index. The verification test was also conducted. RESULTS:The contents of chlorogenic acid and rutoside in 10 batches of S. involucrate were 0.380%-0.546% and 0.334%-0.617%;the optimal decoction technology was as follows as the amout of crude material of S. involucrate 100 g,soaking for 20 min,decocting for 3 times,12,10 and 10 fold of water,decocting 45,30 and 30 min,respectively. The extraction transfer rates of chlorogenic acid and rutoside were 96.2%(RSD=2.66%,n=3)and 89.3%(RSD=3.31%,n=3)in verification test. CONCLUSIONS:For S. involucrate from different producing areas,the contents of effective components are different;optimized decoction and extraction technology is stable and feasible.
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Objective To investigate the therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis (SAP) in rats.Methods A total of 80 healthy Sprague-Dawley rats were divided into eight groups:SAP group (three hours、48 hours),Saussurea involucrate treated group (three hours、48 hours),ulinastatin control group (three hours、48 hours) and sham operation group (three hours、48 hours),10 rats in each group.After modeling,the rats of SAP group were regularly feeded and the rats of other three group were treated with Saussurea involucrate injection (1.04 mL/kg) intraperitoneal injection,ulinastatin 10 000 U/L tail vein injection,and saline femoral vein injection,respectively and injected every 12 hours.At three hours and 48 hours after treated,blood and pancreatic tissue samples were obtained.The mortality rate,serum amylase level and pathological changes of the pancreas of each group were observed.Serum tumor necrosis factor alpha (TNF-α),interleukin (IL)-6 and IL-10 levels were detected by enzyme linked immunosorbent assay (ELISA).The content of malondialdehyde (MDA) in pancreatic tissues was determined by chemical colorimetry.The level of TNF-α mRNA,IL 6 mRNA and IL-10 mRNA in pancreatic tissues were measured with reverse trascription-polymerase chain reaction (RT-PCR).The activity of nuclear factor kappa B p65 (NF-κB p65) in the pancreatic tissue was tested by immunohistochemistry.Single factor analysis of variance was used to compare multiple groups,and the least significant difference (LSD) method was used in the multiple comparisons between groups.Fisher's exact probability method was performed for rates comparison.Results At 48 hours,there was no statistically significant difference in mortality rate among Saussurea involucrate treated group,SAP group and ulinastatin groups (all P＞0.05).At 48 hours,the histopathology score (8.13 ± 0.64),levels of serum amylase ((2 597.0±214.0) U/L),TNF-α ((254.4±11.6) ng/L),IL-6 ((441.4±14.6) ng/L),levels of pancreatic tissues MDA ((311.0±10.6) mmol/L),TNF-α mRNA(2.04±0.08),IL-6 mRNA (1.77±0.04)and activity of NF-κB p65 ((25.90±2.90)％) of Saussurea involucrate treated group were all lower than those of SAP group (11.40±0.89,(4 780.0±101.0) U/L,(396.0±7.4) ng/L,(664.4± 7.6) ng/L,(418.0± 10.6) mmol/L,2.94±0.03,2.63±0.08 and (51.60±5.27) ％;however level of serum IL-10 ((133.5±6.9) ng/L vs (95.1±5.2) ng/L) and IL-10 mRNA of the pancreatic tissue (1.38±0.06 vs 0.85±0.03) significantly increased (F=253.07、441.63、489.40、2 465.00、196.65、477.89、562.79、131.70、560.18、570.04,all P＜0.01).There was no significant differences in all above parameters between Saussurea involucrate treated group and ulinastatin groups (7.56±0.88,(2 607.0±239.0) U/L,(252.2 ±9.2) ng/L,(443.4±9.6) ng/L,(308.4±9.2) mmol/L,2.10±0.12,1.74±0.04,(26.00±3.67)％,(134.5±7.8) ng/L and 1.42±0.06) at 48 hours (all P＞0.05).Conclusion Saussurea involucrate injection can eliminate oxygen free radicals and prevent to xidation,inhibit NF-κB activation,regulate synthesis and release of cytokines,and alleviate pancreatic injury in SAP rats,but it can not decrease mortality.
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Objective To investigate the effects of Saussurea involucrate by cell culture on increasing the calcium content and bone density after 90 days feed in ovariectomized rats. Methods Osteoporosis model rats were induced by ovariectomy, treated with 16.7、33.3、100 mg/(kg?d)cell culture of Saussurea involucrate for 90 days. Body weight was observed, calcium content was measured through atomic absorption method and the bone density was detected by bone sonometers. Results cell culture of Saussurea involucrate(16.7、33.3、100 mg/kg)can increase the calcium content[(380.1 ± 1.9)mg/g、(370.7 ± 1.1)mg/g、(363.5 ± 2.4)mg/g] significantly(P<0.01)than the model group; increase distal femoral bone mineral density[(0.041 ± 0.017)g/cm2、(0.042±0.023)g/cm2、(0.040±0.008)g/cm2] significantly(P<0.01 or 0.05) than the model group(0.022±0.014)g/cm2; and also increase the middle femoral bone mineral density, the low-dose group (0.305±0.030)g/cm2 significantly (P<0.05) than the model group(0.259±0.061)g/cm2. Conclusion The cell culture of Saussurea involucrate has the effect of increasing the bone density.
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Objective:The tissue culture of Saussurea involucrata has been studied for the protection of immune function activities in animal models in our research.Methods: The tissue culture of S.involucrata at the doses of 75,150 and 300mg/kg were given to mice by intragastric administration,successive medication for 7 days,and then the effects of S.involucrata were investigated in mice by carbon clearance rate,DNCB induced delayed hypersensitivity and serum hemolysin formation.Results: The tissue culture of S.involucrata at the doses of 150mg/kg i.g.,for 7 days could inhibit the non-specificity of immune function in mice.At the doses of 300mg/kg i.g.,for 7 days it could have significantly inhibitory effect on delayed hypersensitivity in mice and increase the humoral immunity activity in mice.Conclusion: The tissue culture of S.involucrata had the protection effect on immune function activities.