Kirenol exerts a potent anti-arthritic effect in collagen-induced arthritis by modifying the T cells balance.

Rheumatoid arthritis is characterized by the imbalance of T cells, which leads to increased pro-inflammatory and reduced anti-inflammatory cytokines. Modulating the balance among T cells is crucial for the treatment of RA. Kirenol is a major diterpenoid components of Herba Siegesbeckiae, which has been applied for arthritic therapy for centuries. Since prior research showed Kirenol exhibited anti-inflammatory effect in rats, in this study we have evaluated the effect and mechanism of bioactive Kirenol in a rat model of collagen-induced arthritis (CIA) on modulation of T cells. After immunization with bovine type II collagen (CII), Wistar rats were orally administered saline (CIA group), 2 mg/kg Kirenol or 2 mg/kg prednisolone daily for 30 days. The severity of arthritis was clinically and histologically assessed. The numbers of CD4⁺CD25⁺Foxp3⁺ T regulatory cells (Tregs) and IFNγ⁺CD4⁺ and IL4⁺CD4⁺ T cells were determined by flow cytometry, the mRNA expression level of Foxp3 was quantified by RT-PCR, cytokine levels were measured by ELISA and CII-induced cell proliferation was quantified in vitro. Kirenol significantly delayed the occurrence and reduced the disease severity of CIA. Histological analysis confirmed Kirenol suppressed joint inflammation and inhibited cartilage and bone destruction, compared to the CIA group. Kirenol also upregulated the mRNA expression of Foxp3, increased the numbers of CD4⁺CD25⁺Foxp3⁺ and IL4⁺CD4⁺ T cells, and reduced the number of IFNγ⁺CD4⁺ T cells. Kirenol reduced the levels of TNF-α, IL-17A and IL-6 in synovial fluid and TNF-α, IL-17A and IFN-γ in serum, and increased the serum levels of IL-4, IL-10 and TGF-ß1. In addition, Kirenol inhibited the ability of CII to induce splenocyte, PBMC and lymph node cell proliferation in vitro, compared to cells from CIA rats. In conclusion, these results suggest that Kirenol may be a potential immunosuppressant for the treatment for rheumatoid arthritis.

[Comparative study on anti-inflammatory and antipyretic effects between Dao-di herb and non Dao-di herb of Huangqin].

OBJECTIVE: To do some comparative study on anti-inflammatory and antipyretic effects between the Dao-di herb and non Dao-di herb of Huangqin (the roots of Scutellaria baicalensis) and provide thinking and evidence for study on geoherbalism and clinical usage of Huangqin. METHOD: The anti-inflammatory action was assessed by auricular swelling induced by dimethylbenzene in mice and the antipyretic action was monitored by dried yeast-induced mice fever. RESULT: All samples of both Dao-di and non Dao-di herbs of Huangqin showed antipyretic effect. The Dao-di Huangqin samples showed antipyretic effect between 61% to 53% , whereas the non Dao-di Huangqin samples between 53% to 43%. Six Dao-di Huangqin samples showed better antipyretic effect than four non Dao-di Huangqin samples. All samples of both Dao-di and non Dao-di Huangqin showed anti-inflammatory effect. Dao-di Huangqin showed anti-inflammatory effect between 73% to 54%, whereas non dao-di Huangqin between 53% to 34%. Six Dao-di Huangqin showed better anti-inflammatory effect than four non Dao-di Huangqin. In totality, results from analysis of geoherbalism showed that geoherbal production areas of Huangqin had better effect than that of the non geoherbal production areas in anti-inflammatory and antipyretic effects. CONCLUSION: Both the Dao-di and non Dao-di Huangqin have effects of anti-inflammatory and antipyretic to a certain extent, but the efficacy of the Dao-di Huangqin surpass the non Dao-di Huangqin.

Kirenol upregulates nuclear annexin-1 which interacts with NF-κB to attenuate synovial inflammation of collagen-induced arthritis in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Kirenol is a diterpenoid compound purified from the Chinese Herba Siegesbeckiae. Siegesbeckiae has been employed for the treatment of arthritis for centuries, its safety and efficacy are documented through a long history of human use. AIM OF THE STUDY: To investigate the effects on collagen-induced arthritis (CIA) and anti-inflammatory mechanism of kirenol. MATERIALS AND METHODS: Kirenol was administrated intragastrically in rats after the onset of CIA. Pathological changes were evaluated by paw swelling and histopathology. Concentration of IL-1ß in synovial fluid and adrenal corticotropin (ACTH) in plasma were determined by Elisa. Western blot was performed to detect the expression of annexin-1 and glucocorticoid receptor alpha (GRα) in synovium. NF-κB DNA binding activity was assessed by electrophoretic mobility shift assays (EMSA). RESULTS: Kirenol (1, 2, and 4 mg/kg) and prednisolone depressed paw swelling and reduced IL-1ß of synovial fluid in the CIA rats (p<0.05 or p<0.01). Kirenol and prednisolone upregulated nuclear annexin-1 and inhibited NF-κB activity in synovium of CIA. The inhibitory effect of kirenol and prednisolone on NF-κB activity was enhanced by anti-annexin-1 Ab. Prednisolone, but not kirenol, downregulated plasma ACTH and GRα expression significantly (p<0.01). CONCLUSION: Kirenol and prednisolone can upregulate nuclear annexin-1 which interacts with NF-κB to inhibit NF-κB activity, reduce cytokines expression and thereby attenuate inflammation of CIA joints. Kirenol does not lead to ACTH or GR downregulation, which is in contrast to classic glucocorticoid prednisolone. Kirenol shares with GCs similar anti-inflammatory mechanism but bypass the considerable limitation of GCs treatment.

A rapid and simple RP-HPLC method for quantification of kirenol in rat plasma after oral administration and its application to pharmacokinetic study.

A rapid and simple reverse-phase high-performance liquid chromatography (RP-HPLC) was developed and validated for the quantification of kirenol in rat plasma after oral administration. Kirenol and darutoside (internal standard, IS) were extracted from rat plasma using Cleanert™ C(18) solid-phase extraction (SPE) cartridge. Analysis of the extraction was performed on a Thermo ODS-2 Hypersil C(18) reversed-phase column with a gradient eluent composed of acetonitrile and 0.1% phosphoric acid. The flow rate was 1.0 mL/min and the detection wavelength was set at 215 nm. The calibration curve was linear over the range of 9.756-133.333 µg/mL (r(2) = 0.9991) in rat plasma. The lower limits of detection and quantification were 2.857 and 9.756 µg/mL, respectively. The intra- and inter-day precisions (relative standard deviation, RSD) were between 2.24 and 4.46%, with accuracies ranging from 91.80 to 102.74%. The extraction recovery ranged from 98.16 to 107.62% with RSD less than 4.81%. Stability studies showed that kirenol was stable in preparation and analytical process. The present method was successfully applied to the pharmacokinetic study of kirenol in male Sprague-Dawley rats after oral administration at a dose of 50 mg/kg.

[Progress of researches on pharmacokinetics of active components of Chinese drugs].

The pharmacokinetic research of Chinese drugs is still in the exploratory stage so far, a great progress has achieved in the researches on active components of them since the 1980s. The progresses in pharmacokinetic research of active components of some commonly used Chinese drugs were reviewed in this paper, and the problems to be solved in future were pointed out.

[Effect of Danzhi Xiaoyao Powder on neuro-immuno-endocrine system in patients with depression].

OBJECTIVE: To observe the effect of Danzhi Xiaoyao Powder (DXP) on neuro-immuno-endocrine system in patients with depression. METHODS: A randomized double-blinded and controlled study was conducted in 63 cases of depression. They were assigned to the DXP group (32 cases, treated with DXP) and the control group (31 cases, treated with maprotiline). The curative effect was evaluated with Hamilton's depressive scale (HAMD) before and at the end of the 2nd, 4th and 6th week of the treatment. Serum levels of serotonin (5-HT), norepinephrine (NE), brain derived neurotrophic factor (BDNF), cortisol (CORT), interleukin-6 (IL-6), and interleukin-1beta (IL-1beta) were detected before and at the 6th week of the treatment. RESULTS: After 2 weeks of treatment, the total score of HAMD decreased remarkably in both groups (P < 0.01), and the total score, as well as the scores of the three factors, i. e. anxiety/somatization, cognitive impairment and feeling of despair, were lower in the DXP group than that in the control group respectively (P < 0.05 or P < 0.01). After 4 and 6 weeks of treatment the total score and score of the three factors all reduced significantly in both groups (P < 0.01), with insignificant difference between the groups. After 6 weeks of treatment, the serum levels of 5-HT and BDNF increased (P < 0.01), and the serum IL-6 level decreased in both groups (P < 0.05 or P < 0.01), the serum CORT level reduced in the DXP group (P < 0.01), while the serum NE level elevated in the control group (P < 0.01). CONCLUSION: DXP is effective in improving symptoms of depression by regulating the levels of 5-HT, BDNF, CORT and IL-6.

[Clinical observation on effect of danzhi xiaoyao powder in treating depression].

OBJECTIVE: To observe the effect and side effect of Danzhi Xiaoyao powder (DXP) in treating depression. METHODS: A randomized controlled and double-blinded study was conducted in 63 cases of depression by divided them into the western medicine group (WMG, 31 cases) treated with maprotiline, and the Chinese medicine group (CMG, 32 cases) treated with DXP. The effect of therapy was evaluated before and at the 2nd, 4th and 6th week of the treatment with Hamilton's depressive scale (HAMD), self-rating depression scale (SDS), self-rating anxiety scale (SAS) and the scale for TCM syndrome and symptom differentiation (TCM-SSD), and the side-effect of therapy was assessed with Asberg side-effect scale as well. RESULTS: There was no significant difference between the two groups in scores of HAMD, SDS, SAS, and TCM-SSD. The markedly effective rate in CMG was 84% and in WMG 87%, showed no significance between them (P > 0.05). The scores of HAMD, SDS and SAS of both groups were remarkably lowered after therapy (P < 0.05). However, the score of Asberg in CMG was lower than that in WMG (P < 0.05). CONCLUSION: DXP shows the effect equivalent to that of maprotiline, but with obviously less side-effect.

[Study on free radical mechanism of guanxin II in antagonizing ischemic myocardial damage].

OBJECTIVE: To study comparatively the effect of Guanxin II and its constituents, including Salvia, Red Peony root, Chuanxiong, Safflower and Dalbergia wood, in scavenging active oxygen free radical (OFR) to explore their mechanism in overcoming the experimental acute ischemic myocardial injury and protecting myocardial tissue. METHODS: The experimental acute myocardial ischemic model was established by intraperitoneal injection of pituitrin to rats. OFR level in animal heart tissue was directly measured with low-temperature electron paramagnetic resonance (EPR) spectrometer. RESULTS: The pathological examination of HE stained slide of myocardial tissue and electrocardiography of model animal showed that typical changes of acute myocardial ischemia occurred in myocardial tissue, EPR showed that OFR level in myocardial tissue increased abnormally. The ethanol extract of Guanxin II and its constituents could lower the increased OFR level close to normal, thus to alleviate the myocardial damage. CONCLUSION: Overproduction of OFR could induce damage of heart tissue, its level could be measured directly using low temperature EPR. One of the molecular mechanisms of Guanxin II and its constituents in antagonizing and repairing myocardial damage is to scavenge the abnormal increased active OFR in tissue. This study has provided a basis for further studying the mechanism of Chinese composite recipes and their constituents.