Synergistic anti-inflammatory effects of silibinin and thymol combination on LPS-induced RAW264.7 cells by inhibition of NF-κB and MAPK activation.

BACKGROUND: Combination drug therapy has become an effective strategy for inflammation control. The anti­inflammatory capacities of silibinin and thymol have each been investigated on its own, but little is known about the synergistic anti-inflammatory effects of these two compounds. PURPOSE: This study aims to investigate the synergistic anti-inflammatory effects of silibinin and thymol when administered in combination to lipopolysaccharide (LPS)-induced RAW264.7 cells. METHODS: RAW264.7 cells were pre-treated with silibinin and thymol individually or in combination for 2 h before LPS stimulation. Cell viability was detected by the MTT assay. Nitric oxide (NO) production was measured by Griess reagent. Reactive oxygen species (ROS) was evaluated by 2',7'-dichlorofluorescein-diacetate. ELISA was used to detect tumour necrosis factor-α (TNF-α), and interleukin-6 (IL-6). Western blot was performed to analyse the protein expression of LPS-induced RAW264.7 cells. RESULTS: We observed a synergistic anti-inflammatory effect of silibinin and thymol when administered in combination to LPS-induced RAW264.7 cells. Silibinin combined with thymol (40 µM and 120 µM respectively, with the molar ratio 1:3) had more potent effects on the inhibition of NO, TNF-α, and IL-6 than those exerted by individual administration of these compounds in LPS-induced RAW264.7 cells. The combination of silibinin and thymol (40 µM and 120 µM respectively, with the molar ratio 1:3) strongly inhibited ROS and cyclooxygenase-2 (COX-2). More importantly, the combination of silibinin and thymol (40 µM and 120 µM respectively, with the molar ratio 1:3) was also successful in inhibiting nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) activities. Our results suggest that the synergistic anti-inflammatory effects of silibinin with thymol were associated with the inhibition of NF-κB and MAPK signalling pathways. CONCLUSION: The combination of silibinin and thymol (40 µM and 120 µM, respectively, with the molar ratio 1:3) could inhibit inflammation by suppressing NF-κB and MAPK signalling pathways in LPS-induced RAW264.7 cells.

[Changes of Main Syndromes of Gan Failing to Maintain Normal Flow of Qi in Premenstrual Dysphoric Disorder].

Objective To observe the changes of Chinese medicine (CM) symptoms, the distri- bution characteristics of CM syndromes, and related neuroendocrine levels in premenstrual dysphoric disorder (PMDD) patients. Methods Totally 3 541 female outpatients (18 -45 years old) were inter- viewed by clinical epidemiological questionnaire. According to PMDD diagnostic criteria in DSM-IV , PMDD patients' CM syndromes were identified. Their scores of main symptoms and CM symptoms of common CM syndromes were compared. Contents of 8 neuroendocrine indicators in serum were detected [5- hydroxytryptamine (5-HT) , adrenocorticotropic hormore (ACTH) , angiotensin-II (Ang-II ) , glucocorti- coid (GC), homocysteine (Hcy), melatonin (MLT), nitrogen monoxide (NO), neuropeptide Y (NPY)]. Results Totally 258 PMDD were detected in 3 541 female outpatients (18 -45 years old). The main syn- drome and common syndromes of PMDD patients were reversed invasion of Gan qi syndrome [40.3% (104/258)] and stagnation of Gan qi syndrome [34. 9% (90/258) ], followed by Gan stagnation Pi deficiency syndrome [7.8%(20/258)], Gan stagnation blood stasis syndrome [7.4% (19/258)], Gan stagnation induced fire hyperactivity syndrome [ 6.2% ( 16258 )], Gan stagnation Shen deficiency syndrome [3.1%(8/258)], and Pi-Shen yang deficiency syndrome [0.4%(1258)]. Compared with reversed invasion of Gan qi syndrome, emotional depression and low spirits were main symptoms of stagnation of Gan qi syndrome. Scores for the two symptoms increased (P <0.05). Compared with stagnation of Gan qi syn- drome, irritability, upset, abdominal distension, anxiety, headache, dizziness, insomnia, head distension, bitter mouth, unclear vision were main symptoms of reversed invasion of Gan qi syndrome. Scores for the 10 symptoms increased (P <0.05). Compared with stagnation of Gan qi syndrome, the Hcy level in serum obviously decreased in reversed invasion of Gan qi syndrome (P <0.05). There was no statistical difference in the rest indices (P >0. 05). Conclusions PMDD is closely related to Gan failing to maintain normal flow of qi. Reversed invasion of Gan qi syndrome and stagnation of Gan qi syndrome have different scientif- ic connotations and biological bases. So regulating Gan should be considered as the first choice.