The protective effects and mechanism of Ruyi Zhenbao Pill, a Tibetan medicinal compound, in a rat model of osteoarthritis.

ETHNOPHARMACOLOGICAL RELEVANCE: Ruyi Zhenbao Pill (RZP) is a prescribed Tibetan formulation for the treatment of white-pulse-disease, yellow-water-disease as well as pain-related disease. RZP is composed of 30 medicinal materials including herbal medicine, animal medicine and mineral medicine. They are widely used in the Tibetan area to treat cerebrovascular disease, hemiplegia, rheumatism, and pain diseases for centuries. AIM OF THE STUDY: The aim of the present study was to evaluate the anti-osteoarthritis function of RZP and to clarify the underlying mechanisms. MATERIALS AND METHODS: The active components in RZP were identified using HPLC methods. Osteoarthritis (OA) animal model was established via intra-articular injection of papain in rat knees. After the administration of RZP (0.45, 0.9 g/kg) for 28 days, the clinical observation was conducted, and pathological changes as well as serum biochemical indexes were detected. Moreover, therapeutic targets and pathways of RZP were discussed. RESULTS: The results showed that RZP could suppress knee joint swelling and arthralgia, thus relieving joint pain and inflammation in OA rats. Microcomputed tomography (µCT)-based physiological imaging and staining pictures confirmed the therapeutic effects of RZP on OA symptoms including knee joint swelling and structural changes with progressive inflammation in OA rats. RZP could promote the synthesis or inhibit the degradation of COLⅡ, attenuate OA-induced OPN up-regulation and thus relieve the OA symptom. Furthermore, RZP (0.45-0.9 g/kg) could all ameliorate the imbalance of biomarkers related to OA such as MMP1, TNF-α, COX2, IL-1ß and iNOS in knee joints or serum. CONCLUSION: In conclusion, RZP could effectively relieve inflammatory reaction induced by OA injury and the formulation could be applied to the treatment of OA therapy.

Systems pharmacology-based dissection of mechanisms of Tibetan medicinal compound Ruteng as an effective treatment for collagen-induced arthritis rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Compound Ruteng (CRT) is a prescribed formulation based on the theory of Tibetan medicine for the treatment of yellow-water-disease. It is consisted with 7 medicinal material include Boswellia carterii Birdw (named "Ruxiang" in Chinese); Tinospora sinensis (Lour.) Merr. (named "Kuan-Jin-Teng" in Chinese), Cassia obtusifolia L (named "Jue-Ming-Zi" in Chinese); Abelmoschus manihot (L.) Medic (named "Huang-Kui-Zi" in Chinese); Terminalia chebula Retz. (named "He-Zi" in Chinese); Lamiophlomis rotata (Benth.) Kudo (named "Du-Yi-Wei" in Chinese) and Pyrethrum tatsienense (Bur. et Franch.) Ling (named "Da-Jian-Ju" in Chinese). They are widely distributed in Tibet area of China and have been used to treat rheumatism, jaundice, and skin diseases for centuries. AIM OF THE STUDY: The present study was conducted to investigate the anti-arthritis effect of CRT and to disclose the systems pharmacology-based dissection of mechanisms. MATERIALS AND METHODS: The chemical constituents in CRT were identified using HPLC method, and CRT candidate targets against RA were screened by network pharmacology-based analysis and further experimentally validated based on collagen-induced arthritis (CIA) rat model. Furthermore, therapeutic mechanisms and pathways of CRT were investigated. RESULTS: 391 potential targets (protein) were predicted against 92 active ingredients of 7 medicinal materials in CRT. Enrichment analysis and molecular docking studies also enforced the practiced results. X-ray based physiological imaging showed the attenuated effect of CRT on paw swelling, synovial joints and cartilage with improved inflammation in CIA rats. Moreover, the expression of biomarkers associated with RA such as MMP1, MMP3 and MMP13 and TNF-a, COX2 and iNOS are down-regulated in ankle joints, serum, or liver. CONCLUSION: In conclusion, CRT compound could attenuate RA symptoms and active ingredients of this compound could be considered for drug designing to treat RA.

Sodium aescinate and its bioactive components induce degranulation via oxidative stress in RBL-2H3 mast cells.

Sodium aescinate (SA) is a vital salt of sodium escin from Aesculus wilsonii Rehd seeds. SA injection (SAI) has received great success in treating cerebral edema, venous reflux disease and other inflammatory conditions. Recently, high incidences of immediate hypersensitivity reactions were reported after SA infusion, which raised questions on safety and risk associated with its clinical application. This study was designed to check whether SAI and its four components induce degranulation using RBL-2H3 mast cells. For this purpose, we evaluated different treatment levels of SAI (20, 40, 60, 80 and 100 µg ml-1) and its four characteristic components, SA-A, SA-B, SA-C and SA-D, at 60 µg ml-1 in different tests including cell viability test, ß-hexosaminidase and histamine assays, oxidative stress indices, apoptosis analysis and intracellular calcium ions in RBL-2H3 cells. Our results demonstrated that SAI at 80 µg ml-1 and 100 µg ml-1, and its two components (SA-B and SA-D) at 60 µg ml-1 were responsible for disturbing cell morphology and cell viability, elevated levels of ß-hexosaminidase, histamine, modulation of oxidative stress indices, induced apoptosis and increase in intracellular calcium ions in RBL-2H3 cells, when compared with the control. Our results demonstrated for the first time that SAI was more likely to induce immediate hypersensitivity reactions attributable to degranulation via oxidative stress caused by SA-B and SA-D components. These results would not only be useful for the safety of end user but also for the industry to improve the quality of SA infusion.

Effects of Veratrilla baillonii Extract on Hepatic Gene Expression Profiles in Response to Aconitum brachypodum-Induced Liver Toxicity in Mice.

This manuscript was aimed to explore the hepato-protective effect of water extract of Veratrilla baillonii Franch. (Gentianaceae) (WVBF) on serious hepatic toxicity induced in mice treated with Aconitum brachypodum Diels (Ranunculaceae) at transcriptome level. The physiological and pathological symptoms were evaluated as the markers for hepato toxicity induced by A. brachypodum Diels (CFA) extracted compounds. Moreover, gene chip method was used to compare and investigate the gene expression level of WVBF on CFA induced-liver toxicity to identify the potential target of WVBF and CFA on liver. The results showed that WVBF had a significant detoxification effect on CFA-induced acute hepatic toxicity. There were 130 genes with lower expression and 124 genes expressed at higher rate in CFA treated group as compared with normal control group, while there are 67 genes down-regulated and 74 genes up-regulated in WVBF treated group in comparison with CFA treated group. WVBF could attenuate CFA-induced liver damage in mice through regulating oxidative stress, inflammatory injury and cell apoptosis/necrosis pathways. On the other hand, WVBF and CFA may have potential synergetic effects on the target genes of certain diseases such as inflammation, cancer and diabetes.

[Rotational and vibrational state distributions of CsH in the reactions of Cs(6 2D5/2) with H2].

The nascent quantum state distributions of the CsH product resulting from the reaction Cs(6D5/2) with Hz were determined using a laser pump-probe technique in a five-arm crossed heat-pipe oven. Cs-H2 mixture was irradiated with pulses of 885.4 nm radiation from a OPO laser, populating 6D5/2 state by two-photon absosption. Laser induced fluorescence was used to detect CsH molecules directly at the collision volume by scanning pulse tunable dye laser over X 1 sigma+ (v", J") --> A 1 sigma+ (v', J' = J" +/- 1) absorption line. The vibration bands (v" = 0, v' = 6) and (v" = 1, v' = 9) were chosen. For the investigated reaction, the nascent CsH product molecules were found to populate the lowest two vibrational (v" = 0, 1) levels of the ground electronic state but could not be detected in any higher vibrational state. Rotational distributions of CsH products obtained for v" = 0 and 1 states appear to be monomodal , peaking in J = 6-8. The rotational population profile is roughly consistent with a statistical distribution at the system temperature. A plot of logarithm of relative population of states J divided by the degeneracy factor (2J + 1) against J (J + 1) was yielded. The linearity of the plot establishes the Boltzmann form for rotational distributions of both the v" = 0 and 1. The rotational temperatures are (458 +/- 20) K and (447 +/- 18) K for v = 0 and 1, respectively. The nascent CsH rotational temperatures were found to be slightly below the cell temperature. The relative vibrational population was determined to be 0.527 and 0.473. The average vibrational and rotational energy release can be computed. The relative fractions (fv), (fR) and (fT) of average energy disposal were derived as 0.25, 0.10 and 0.65 respectively, having a major translation energy release. All of the above results support the assumption that the Cs(6 2D5/2)-H2 reaction occurs primarily in a collinear geometry by a harpoon mechanism but not an insertion.

[Measurement of the cross section in Li(2P)+H2-->LiH+H reaction].

At a Li density of approximately 10(13) cm(-3), the lithium vapor was irradiated in a five-arm stainless steel heat pipe oven containing Li and H2 with pulses of radiation from a N2-laser-pumped dye laser, populating Li(2P) state by the Li(2S-->2P) resonance transition at 670.8 nm. Typical operating pressure of H2 was 60-300 Pa. The cross section for Li(2P)+H2-->LiH+H reaction was measured using method of atomic fluorescence. The decay signal of the time-resolved fluorescence from the 2P-->2S transition was monitored. The decay curve of the Li(2P) can be treated as a single exponential function. The effective lifetimes of the 2P state was obtained. According to the Stern-Volmer equation, a plot of reciprocal of effective lifetimes of the 2P state quenched by H2 against its densities yielded a slope that indicated the total cross section for deactivation and an intercept (at which the H2 pressure is zero) that provided the information about the radiative lifetime of the state. The total quenching (reactive+nonreactive) cross section for deactivation of the 2P state by means of collisions with H2 is (25.1 +/- 4.0) x 10(-16) cm2. The reactive cross section could be obtained using results of the recording of the fluorescence signals with rapid rise in transient regime (<10 ns) Li(2P-->2S) at the different H2 densities. The authors fitted a two-state rate equation model to obtain the cross section sigma (Li(2P)+H2-->LiH+H) = (0.2 +/- 0.1) x 10(-16) cm2. The authors' results imply that reactive collisions occur on average 1/125 as often as quenching collisions. The cross section for reaction is small but not negligible.