Bai-Hu-Tang regulates endothelin-1 and its signalling pathway in vascular endothelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Bai-Hu-Tang (BHT) is traditionally used to treat human and animal fever syndrome with four symptoms: large and vigorous pulse, large thirst, high sweat, and high heat. AIM OF THE STUDY: To investigate the mechanism of vasodilation regulation of Bai-Hu-Tang in primary vascular endothelial cells stimulated by lipopolysaccharide (LPS). MATERIALS AND METHODS: A hydrophilic concentrate of BHT was prepared, and the main components of mangiferin and timosaponin BⅡ were determined by HLPC analysis. The rabbit fever model was constructed by intravenous injection of LPS (15 µg/kg body weight), and BHT was gavaged to treat febrile rabbits. After treatment for 6 h, animal peripheral blood was collected, and serum was isolated for endothelin-1 (ET-1) and nitric oxide (NO) assays. Rabbit vascular endothelial cells (RVECs) were isolated and stimulated with 1 µg/mL LPS, and then inflammatory cells were treated with 125 or 250 µg/mL BHT for 24 h. The supernatant cytokines TNF-ɑ, IL-1ß, IL-6, and ET-1 were detected by ELISA kits. Gene expression levels of endothelin receptor type B (ETB receptor) were analysed by real-time polymerase chain reaction (RT-PCR), and protein expression levels of PI3K and Akt were detected by Western blot. A nitrite assay was used to measure intracellular nitric oxide (NO) production, and nitric oxide synthase (NOS) was measured by the T-NOS colorimetric method. RESULTS: Animal experiments demonstrated that BHT significantly restored ET-1 and NO in animal peripheral blood, which were disordered in LPS-induced fever rabbits. Moreover, a cytotoxicity assay demonstrated that BHT ≤700 µg/mL is innoxious to RVECs. BHT significantly repressed cellular TNF-α, IL-1ß, and ET-1, which were originally elevated by LPS in RVECs. Meanwhile, BHT elevated the gene expression level of the ETB receptor and promoted NOS and NO production in RVECs induced by LPS. CONCLUSION: BHT can inhibit excessive ET-1 secretion induced by LPS in vascular endothelial cells and activate the classic ET-1 signalling pathway to promote NO production, which may facilitate vasodilation of smooth muscle cells.

Schisandrin Protects against Norepinephrine-Induced Myocardial Hypertrophic Injury by Inhibiting the JAK2/STAT3 Signaling Pathway.

Aims. Heart failure is closely associated with norepinephrine-(NE-) induced cardiomyocyte hypertrophy. Schisandrin is derived from the traditional Chinese medicine Schisandra; it has a variety of pharmacological activities, and the mechanism of schisandrin-mediated protection of the cardiovascular system is not clear. Main Methods. NE was used to establish a cardiomyocyte hypertrophy model to explore the mechanism of action of schisandrin. An MTT assay was used for cell viability; Hoechst fluorescence staining was used to observe the cell morphology and calculate the apoptosis rate. The cell surface area was measured and the protein to DNA ratio was calculated, changes in mitochondrial membrane potential were detected, and the degree of hypertrophic cell damage was evaluated. WB, QRT-PCR, and immunofluorescence were used to qualitatively, quantitatively, and quantitatively detect apoptotic proteins in the JAK2/STAT3 signaling pathway. Key Findings. In the NE-induced model, schisandrin treatment reduced the apoptosis rate of cardiomyocytes, increased the ratio of the cell surface area to cardiomyocyte protein/DNA, and also, increased the membrane potential of the mitochondria. The expression of both JAK2 and STAT3 was downregulated, and the BAX/Bcl-2 ratio was significantly reduced. In conclusion, schisandrin may protect against NE-induced cardiomyocyte hypertrophy by inhibiting the JAK2/STAT3 signaling pathway and reducing cardiomyocyte apoptosis.

Cardioprotection of Sheng Mai Yin a classic formula on adriamycin induced myocardial injury in Wistar rats.

BACKGROUND: Sheng Mai Yin (SMY), a well-known Chinese herbal medicine, is widely used to treat cardiac diseases characterized by the deficiency of Qi and Yin syndrome in China. SMY-based treatment has been derived from Traditional Chinese Medicine (TCM), officially recorded in the Chinese Pharmacopoeia. PURPOSE: We aimed to clarify whether SMY attenuates myocardial injury induced by adriamycin in Wistar rats with chronic heart failure (CHF). METHODS: To quantify ginsenoside Rg1, ophiopogonin D, ophiopogonin D', schisandrin by HPLC. To establish CHF animal model, adriamycin was intraperitoneally injected in Wistar rats for 7 weeks at a dose of 2 mg/kg body weight. Overall, 180 rats were randomly assigned to six groups: control, CHF model, captopril (positive control), high dose (HSMY), medium dose (MSMY), and low dose (LSMY). Experimental rats were fed 0.625 mg/kg captopril and 90 mg/kg, 45 mg/kg, and 22.5 mg/kg SMY, respectively, over 7 weeks. The inflammatory cytokines TNF-α and IL-6 were measured using ELISA. Matrix metalloproteinases (MMPs) were identified using immunohistochemistry (IHC). Both IHC and RT-PCR were used for quantification of COL-IV expression levels in the heart tissues. Scanning electron microscopy (SEM) was used for the visualization of myocardium morphology. RESULTS: The concentration of ginsenoside Rg1, ophiopogonin D, ophiopogonin D' and schisandrin in SMY was found to be 25.63 ±â€¯3.42 mg, 11.00 ±â€¯1.17 mg, 7.02 ±â€¯0.51 mg, and 25.31 ±â€¯4.28 mg per gram of SMY, respectively. Compared with CHF model group, TNF-α levels were significantly lower (p < .01) in the four drug-administered groups. Moreover, except in the SYM low dose group, IL-6 levels in the other 3 drug-administered groups were also significantly reduced (p < .01). COL-IV expression was also significantly reduced on treatment with high SYM dose (p < .05). IHC results confirmed that SMY and captopril significantly reduced MMPs expression in the heart. CONCLUSION: SMY could control or slow CHF progression by suppressing pathological changes in the myocardium in CHF models. This could be attributed at least partly to the downregulation of IL-6 and TNF-α and inhibition of overexpression of MMPs and COL-IV, which significantly relieved the cardiac-linked pathologies, decreased the risk of myocardial fibrosis, and inhibited cardiac remodeling. These findings suggested that SMY and captopril have similar efficacy for the treatment of adriamycin-induced myocardial injury. In addition, Chinese herbal preparation SMY may play a role in the treatment of cardiac diseases.

Analgesic and anti-inflammatory effects of hydroalcoholic extract isolated from Semen vaccariae.

Semen vaccariae, the seeds of Vaccaria segetalis (Neck.) Garcke, is usually used as an important medication for female mammary gland diseases; it has also been used to promote lactation for centuries in China. The purpose of this work was to evaluate the analgesic and anti-inflammatory effects of hydroalcoholic extract from semen vaccariae (HESV) with oral doses of 50, 100 and 200mg/kg•bw in mice and rats. We observed that the HESV could effectively inhibit acetic acid-induced abdominal contraction and could elevate the latency time to thermal stimuli in the hot-plate test in mice. In the xylene-induced ear-swelling test in mice, HESV could suppress the ear swelling. Additionally, HESV could significantly decrease the peritoneal capillary permeability and leukocyte infiltration in mice induced by the intraperitoneal injection of acetic acid. HESV also significantly reduced paw thickness 2-4 hours after the injection of carrageenan in the carrageenan-induced rat paw edema test. This study was the first to demonstrate that the oral administration of HESV might play an important role in the process of analgesia and anti-inflammation, supporting its use for female mammary gland diseases in traditional medicine.

Efficacy of herbal tincture as treatment option for retained placenta in dairy cows.

Retained placenta remains therapeutic challenge in cattle. Certain traditional medicines are believed to be able to alleviate retained placenta condition and improve overall fertility in cows. The aim of the present study was to evaluate the efficacy of an herbal tincture for treatment of retained placenta. The herbal tincture was extracted from a combination of Herba Leonuri, Angelicae Sinensis Radix, Flos Carthami, Myrrha and Rhizoma Cyperi by percolation with 70% ethanol to a concentration of 0.5g crude herb/ml. Cows diagnosed with retained placenta (n=48) were randomly divided into one of two treatment groups (A and B), with animals in group A (n=26) receiving herbal tincture orally, and cows in group B (n=22) receiving oxytetracycline infusion into the uterus. Eighty six cows with no clinically visible pathological conditions, given birth alone and with no retained placenta diagnosis were included into control group (C). Retained placenta was expelled within 72h following initial treatment in 19 cows in group A, yet no cows in group B were recorded to expel placenta in the same time. The median number of days to first service (70.0 vs. 102.5 days; P<0.05) and median number of days open (76.0 vs. 134.0 days; P<0.01) were lower in group A than in group B. Percentage of cows pregnant within 100 days postpartum was the highest for animals in group A compared to controls (61.5% vs. 39.5%, P<0.05), and for animals in group B (61.5% vs. 22.7%; P<0.01). Herbal tincture used in the present study might facilitate expulsion of retained placenta and improve subsequent fertility, thus could present effective treatment option for retained placenta in cows.

The administration of Sheng Hua Tang immediately after delivery to reduce the incidence of retained placenta in Holstein dairy cows.

Sheng Hua Tang, a classical herbal formula consisting of Radix Angelicae sinensis, Ligustici rhizoma, Semen persicae, Zingiberis rhizoma, and Radix glycyrrhizae, is known to be beneficial in alleviating postpartum diseases and facilitating a return to normal reproductive function. This study investigated whether the administration of Sheng Hua Tang within 2 to 4 hours after delivery was effective as a preventive treatment for reducing the risk of retained placenta in Holstein dairy cows. A total of 357 cows, each of which had delivered its calf spontaneously, were randomly allocated to one of two groups. In the treatment group, the cows (n = 175) received Sheng Hua Tang with an oral dose of 0.36 g crude herb per kg·body weight once daily for three consecutive days. The controls (n = 182) received no treatment. The placental retention proportion was 4.0% and 17.0% within 12 hours after delivery in the treated and control animals, respectively (P < 0.01). We found decreases in the calving-to-first-service interval (73.2 ± 25.1 vs. 81.9 ± 32.8 days; P < 0.01), calving-to-conception interval (93.4 ± 38.8 vs. 114.6 ± 42.9 days; P < 0.01), and service per conception (1.5 ± 0.8 vs. 1.9 ± 1.0 days; P < 0.01) in the treatment group compared with the control group. The first artificial insemination conception proportion was higher in the treatment group than in the control group (60.4% vs. 41.1%; P = 0.01). Moreover, the between-group difference in the proportion of cows that were pregnant within 180 days postpartum approached statistical significance (88.2% vs. 80.6%; P = 0.07). Sheng Hua Tang showed beneficial effects in reducing the incidence of retained placenta and improving subsequent reproductive performance in cows. This preventive treatment strategy would be effective in improving the management of puerperal health. The potential benefits of Sheng Hua Tang warrant further investigation to determine whether this preventive treatment strategy can be endorsed as a general preventive approach in postpartum cows.

Aqueous extract of Bai-Hu-Tang, a classical Chinese herb formula, prevents excessive immune response and liver injury induced by LPS in rabbits.

ETHNOPHARMACOLOGICAL RELEVANCE: Bai-Hu-Tang (BHT) was traditionally used to reduce fever heat and promote generation of body fluids. AIM OF THE STUDY: To investigate the effect and mechanism of BHT in the prevention of lipopolysaccharide (LPS) fever in manners of immune modulation. MATERIALS AND METHODS: The model of fever syndrome of Chinese medicine pattern was imitated by LPS injection i.v. in rabbits, and BHT was gavaged. The serum levels of tumor necrosis factor-α (TNF-α), interleukin (IL-6, 10) and immunoglobulin (IgG, IgA, and IgM) were determined by enzyme-linked immunosorbent assay (ELISA); alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were tested by biochemical methods. Liver tissue damage was detected by hematoxylin-eosin (H&E) stain. Subpopulation of T cells was detected by Fluorescence Activated Cell Sorter (FACS). Genes expression of Toll-like receptor 4 (TLR4) and lipopolysaccharide binding protein (LBP) in liver tissue were assayed by real-time polymerase chain reaction (RT-PCR). RESULT: The results demonstrated that BHT prevented sudden increase of IL-10, TNF-α, ALT and AST, and liver damage induced by LPS. BHT also prevented significant decrease of the percentage of CD(8+) T cells since LPS injection. At the same time, BHT did not affect the gene expression of TLR4 and serum concentration of three immunoglobulins, which were increased by LPS, but made gene expression of LBP higher. CONCLUSION: The results of this study indicated that BHT played an important role in immunity protection and anti-injury through preventing immunoinflammatory damage by LPS. The achievement thereby scientifically provided mechanism of BHT in the prevention of febrile disease, and supported its traditional use.