Effect of vinegar steaming on the composition and structure of Schisandra chinensis polysaccharide and its anti-colitis activity.

In this study, infrared spectroscopy, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) technology were applied to systematically explain the Schisandra chinensis's polysaccharide transformation in configuration, molecular weight, monosaccharide composition, and anti-ulcerative colitis (UC) activity after vinegar processing. Scanning electron microscopic results showed that the appearance of S. chinensis polysaccharide changed significantly after steaming with vinegar. The MALDI-TOF-MS results showed that the mass spectra of raw S. chinensis polysaccharides (RSCP) were slightly lower than those of vinegar-processed S. chinensis polysaccharides (VSCP). The RSCP showed higher peaks at m/z 1350.790, 2016.796, and 2665.985, all with left-skewed distribution, and the molecular weights were concentrated in the range of 1300-3100, with no higher peak above m/z 5000. The VSCPs showed a whole band below m/z 3000, with m/z 1021.096 being the highest peak, and the intensity decreased with the increase of m/z. In addition, compared to RSCPs, VSCPs can significantly increase the content of intestinal short-chain fatty acids (SCFAs). This study showed that the apparent morphology and molecular weight of S. chinensis's polysaccharides significantly changed after steaming with vinegar. These changes directly affect its anti-UC effect significantly, and its mechanism is closely related to improving the structure and diversity of gut microbiota and SCFA metabolism.

Effects of different steaming times on the composition, structure and immune activity of Polygonatum Polysaccharide.

ETHNOPHARMACOLOGICAL RELEVANCE: As a commonly used traditional Chinese herbal medicine, Polygonati Rhizoma has high medicinal value, it can enhance the immune capacity of the body, regulate the metabolism of blood glucose and lipids, treat weakness of the stomach and intestines and physical fatigue, and so on. There are three plant varieties of Polygonati Rhizoma recorded in Chinese Pharmacopoeia, including Polygonatum sibiricum Red., Polygonatum kingianum Coll. et Hemsl. and Polygonatum cyrtonema Hua, compared with the first two, Polygonatum cyrtonema Hua is less studied. Polygonatum cyrtonema Hua is one of the basal plants of the Chinese herb Polygonati Rhizoma, that strengthens the spleen, moistens the lungs, and benefits the kidneys. Polygonatum polysaccharide is the main active substance of Polygonatum cyrtonema Hua, which has various biological effects of regulating immune system, anti-inflammatory, anti-antidepressant, antioxidant and other effects. AIM OF THE STUDY: In order to analyze the necessity and scientificity of multiple cycles of steaming during the traditional nine-steaming and nine-drying process of the concoction of Polygonatum, we investigated the changes in the composition and structure of polysaccharides, and explored its immunomodulatory activity and molecular biological mechanism. METHODS: The structural characterization and molecular weight of polysaccharides were studied by scanning electron microscope (SEM), high-performance size exclusion chromatography-evaporative light scattering detector (HPSEC-ELSD) and Matrix.assisted laser resolutionu ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The composition and proportion of monosaccharides were determined by PMP-HPLC method. A mouse immunosuppression model was established by intraperitoneal injection of cyclophosphamide to compare the immunomodulatory effects and mechanisms of different steaming times of Polygonatum, The changes of body mass and immune organ indices of mice were measured; the secretion levels of interleukin-2 (IL-2), interferon γ (IFN-γ) and the expression levels of immunoglobulin M (IgM) and immunoglobulin A (IgA) in serum were determined by enzyme-linked immunosorbent assay; and then flow cytometry was used to detect T-lymphocyte subpopulations to evaluate the differences of immunomodulatory effects of polysaccharides during the processing and preparation of Polygonatum. Finally, the Illumina MiSeq high-throughput sequencing platform was used to analyze short-chain fatty acids and to investigate the effects of different steaming times of Polygonatum polysaccharides on immune function and intestinal flora in immunosuppressed mice. RESULTS: The structure of the Polygonatum polysaccharide with different steaming times changed significantly, the relative molecular weight of Polygonatum polysaccharide decreased significantly, and the monosaccharide composition of Polygonatum cyrtonema Hua with different steaming times was the same but the content was different. The immunomodulatory activity of the Polygonatum polysaccharide was enhanced after concoction, which significantly increased the spleen index and thymus index, and increased the expression of IL-2, IFN-γ, IgA and IgM. The CD4+/CD8+ ratio of Polygonatum polysaccharide also increased gradually with different steaming times, indicating enhanced immune function and significant immunomodulatory effect. The content of short-chain fatty acids in the feces of mice in both six steaming six sun-drying of Polygonatum polysaccharides (SYWPP) and nine steaming nine sun-drying of Polygonatum polysaccharides (NYWPP) groups increased significantly, including the content of propionic acid, isobutyric acid, valeric acid, and isovaleric acid, and also had a good effect on the regulation and improvement of microbial community abundance and diversity, SYWPP and NYWPP increased the relative abundance of Bacteroides and the ratio of Bacteroides and Firmicutes (B:F), while SYWPP significantly increased the abundance of Bacteroides, Alistipes and norank_f__Lachnospiraceae, but the effect of raw Polygonatum polysaccharides (RPP) and NYWPP was not significant than SYWPP. CONCLUSION: Overall, both SYWPP and NYWPP could significantly enhance the immune activity of the organism, improve the imbalance of intestinal flora in immunosuppressed mice, and increase the content of intestinal short chain fatty acids (SCFAs), it is noteworthy that SYWPP has a better effect on the improvement of the immune activity of the organism. These findings can explore the stage of the concoction process of Polygonatum cyrtonema Hua to achieve the best effect, provide a reference basis for the development of quality standards, and at the same time promote the application of new therapeutic agents and health foods in raw and different steaming times of Polygonatum polysaccharide.

The multi-omics and analysis to reveal thermal processing enhanced anti-rheumatoid arthritis efficacy of Radix Clematidis in rats.

Radix Clematidis (RC) is the roots and rhizomes of Clematis chinensis Osbeck, which has potent effects for expelling wind and dispelling dampness. Processing RC with yellow wine is a traditional processing method. This study aimed to investigate thermal and yellow wine processing influences on potential effective components of RC and its anti-rheumatoid arthritis enhancement mechanisms. Different thermal and wine processing were adopted to get different RC samples. Scanning electron microscope and mercury intrusion porosimetry were used to measure fractal parameters of pore structure. Based on ultra performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF/MS), main constituents were identified and quantified. Besides, the correlation between fractal parameters and main constituents was investigated. The anti-rheumatoid arthritis effect was researched in adjuvant-induced arthritis (AIA) rats. The levels of inflammatory cytokine were determined with ELISA kits. Non-targeted serum metabolomics was performed with UPLC-QTOF/MS. 35 compounds were identified in RC, mainly triterpenoid saponins, also including organic acids and lignanoids. The extraction yield of four active triterpenoid saponins significantly increased because looser tissue and wider pore size distribution. Fractal dimension and total surface area significantly increased while total porosity and total volume decreased. In AIA rats, thermal and wine processed RC could markedly inhibit inflammatory cytokines IL-6, IL-1ß, TNF-α, and VEGF. Besides, tryptophan and lipid metabolism disorders were ameliorated. Thermal and yellow wine treatments engendered complex pore structure to increase the contents of four active triterpenoid saponins of RC, leading to greater anti-rheumatoid arthritis efficacy.

[Optimization of processing technology of wine-processed Polygonati Rhizoma based on orthogonal test and AHP-comprehensive scoring method and comparison of immunomodulation functions of Polygonati Rhizoma processed with different methods].

This study employed orthogonal design and AHP-comprehensive scoring method to optimize the processing technology of wine-processed Polygonati Rhizoma, and then explored the immunomodulation performance of the product. Orthogonal test was established based on single factor test results to study the effects of soaking time, steaming time, and drying temperature on the quality of wine-processed Polygonati Rhizoma. Further, the analytic hierarchy process(AHP) and comprehensive scoring method were employed to determine the optimum processing parameters. The immunosuppression model of mice was established by injecting cyclophosphamide intraperitoneally. The body weight, immune organ index, and white blood cell count(WBC) and red blood cell count(RBC) in peripheral blood were compared between the mice administrated with the non-processed Polygonati Rhizoma and the wine-processed Polygonati Rhizoma prepared with modern and traditional methods. Further, the levels of interleukin-2(IL-2) and interferon-gamma(IFN-γ) in serum were determined by enzyme-linked immunosorbent assay(ELISA) for comparison. The processing parameters were optimized as follows: soaking in Chinese rice wine for 10 h, steaming for 20 h, and drying thick slices at 60 ℃. The wine-processed Polygonati Rhizoma prepared with both modern and traditional methods can significantly enhance the immune function, with similar performance. The optimized processing technology of wine-processed Polygonati Rhizoma is stable and feasible and the product prepared with this process has obvious immune-enhancing effect, which provides a basis for the quality standard formulation and the modern research of wine-processed Polygonati Rhizoma.

Shen-Fu Injection () alleviates post-resuscitation myocardial dysfunction by up-regulating expression of sarcoplasmic reticulum Ca(2+)-ATPase.

OBJECTIVE: To compare the effect of Shen-Fu Injection (SFI) and epinephrine on the expression of sarcoplasmic reticulum Ca(2+) ATPase 2a (SERCA2a) in a pig model with post-resuscitation myocardial dysfunction. METHODS: Ventricular fibrillation (VF) was electrically induced in Wu-zhi-shan miniature pigs. After 8 min of untreated VF and 2 min of cardiopulmonary resuscitation (CPR), all animals were randomly administered a bolus injection of saline placebo (SA group, n=10), SFI (0.8 mg/kg, SFI group, n=10) or epinephrine (20 µg/kg, EPI group, n=10). After 4 min of CPR, a 100-J shock was delivered. If the defibrillation attempt failed to attain restoration of spontaneous circulation (ROSC), manual chest compressions were rapidly resumed for a further 2 min followed by a second defibrillation attempt. Hemodynamic variables were recorded, and plasma concentrations of catecholamines were measured. Adenylate cyclase (AC), cyclic adenosine monophosphate (cAMP) and the expressions of ß1-adrenoceptor (AR) and SERCA 2a were determined. RESULTS: Cardiac output, left ventricular dp/dtmax and negative dp/dtmax were significantly higher in the SFI group than in the SA and EPI groups at 4 and 6 h after ROSC. The expression of ß1-AR and SERCA2a at 24 h after ROSC were significantly higher in the SFI group than in the SA and EPI groups (P<0.05 or P<0.01). CONCLUSIONS: The administration of epinephrine during CPR decreased the expression of SERCA2a and aggravated postresuscitation myocardial function (P<0.01). SFI attenuated post-resuscitation myocardial dysfunction, and the mechanism might be related to the up-regulation of SERCA2a expression.

Comparison of Shenfu Injection () and epinephrine on catecholamine levels in a porcine model of prolonged cardiac arrest.

OBJECTIVE: To compare the effects of Shenfu Injection (SFI) and epinephrine (EPI) on catecholamine levels in a porcine model of prolonged cardiac arrest (CA). METHODS: After 8 min of untreated ventricular fibrillation, 24 Wuzhishan miniature pigs were randomly assigned to one of the three groups (n=8 per group) and received central venous injection, respectively: SFI group (1 mL/kg), EPI group (20 µg/kg EPI), and normal saline (NS) group. Cardiac output (CO), maximum rate of increase/decrease in left ventricular pressure (±dp/dt), serum levels of EPI, norepinephrine (NE), and dopamine (DA) were determined at baseline and at 0.5, 1, 2, and 4 h after restoration of spontaneous circulation. RESULTS: The duration of cardiopulmonary resuscitation was shorter in the EPI and SFI groups than in the NS group (P<0.05). The EPI level increased significantly after restoration of spontaneous circulation (ROSC) in all three groups, and was significantly different between the EPI group and the other two groups immediately after ROSC (both P<0.01), but these differences gradually disappeared over time. There were no significant differences in NE or DA levels among the three groups, and there were no correlations between catecholamine levels and CO or dp/dt (P>0.05). CONCLUSIONS: SFI did not significantly affect endogenous catecholamine levels during cardiopulmonary resuscitation after prolonged ventricular fibrillation. However, SFI improved oxygen metabolism, and produced a better hemodynamic status compared with EPI. SFI might be a potentially vasopressor drug for the treatment of CA.

Therapeutic effects of Shenfu Injection on post-cardiac arrest syndrome.

Survival rates after cardiac arrest have not changed substantially over the past 5 decades. Postcardiac arrest (CA) syndrome (PCAS) is the primary reason for the high mortality rate after successful restoration of spontaneous circulation (ROSC). Intravenous administration of Shenfu Injection (, SFI) may attenuate post-CA myocardial dysfunction and cerebral injury, inhibit systemic ischemia/reperfusion responses, and treat underlying diseases. In this article, we reviewed the therapeutic effects of SFI in PCAS. SFI might be useful in the treatment of PCAS, incorporating the multi-link and multi-target advantages of Chinese medicine into PCAS management. Further experimental and clinical research to verify the therapeutic effects of SFI in PCAS is required.

[Effects of 1, 25(OH)(2);D(3); on parathyroid hormone induced transdifferentiation and TGF-ß(1); expression in cultured human renal tubular epithelial cells].

AIM: To explore the effects of 1, 25(OH)(2);D(3); on parathyroid hormone (PTH) induced transdifferentiation and TGF-ß(1); expression in cultured human renal tubular epithelial cells. METHODS: HK-2 cells were cultured in DMEM/F12 medium supplemented with 50 mL/L FBS. Cells were divided into three groups. (1) CONTROL GROUP: without PTH or 1, 25(OH)(2);D(3);; (2) PTH group: 10(-10); mol/L PTH; (3) PTH and 1, 25(OH)(2);D(3); group: 10(-10); mol/L PTH and different concentrations of 1, 25(OH)(2);D(3); (10(-10);, 10(-9);, 10(-8); and 10(-7); mol/L). The gene expressions of α-SMA and TGF-ß(1); were detected by semi-quantitative RT-PCR. The protein expressions of α-SMA and TGF-ß(1); were detected by Western blot. Immunocytochemisty (ICC) was used to measure the expression of α-SMA in HK-2. ELISA was used to assay the level of TGF-ß(1); in the supernatant. RESULTS: The gene expressions of α-SMA and TGF-ß(1); in PTH group were significantly higher than those in control group (P<0.05). In contrast, they were significantly lower in PTH and 1, 25(OH)(2);D(3); group than those in PTH group (P<0.05). Western blot results showed α-SMA could not be detected in normal HK-2 cells, which could be detected in PTH group. TGF-ß(1); protein expression in PTH group was higher than that in control group. In PTH and 1, 25(OH)(2);D(3); group, α-SMA and TGF-ß(1); protein expressions were significantly lower than those in PTH group (P<0.05). ICC results showed that α-SMA was hardly expressed in cells of control group. However, positive expression of α-SMA could be seen in many cells in PTH group. In PTH and 1, 25(OH)(2);D(3); group, the cells of α-SMA positive expressed were significantly less than those in PTH group (P<0.05). ELISA results showed that the level of TGF-ß(1); in the supernatant of PTH group was higher than that in control group, which was also higher than that in PTH and 1, 25(OH)(2);D(3); group (P<0.05). CONCLUSION: 1, 25(OH)(2);D(3); can attenuate PTH-induced transdifferentiation and TGF-ß(1); expression in cultured human renal tubular epithelial cells.