BACKGROUND:Mongolian Pharmaceutical Betel Shisanwei Ingredients Pil has achieved good clinical efficacy, but the underlying mechanism remains unclear. OBJECTIVE: To study the effects of Mongolian Pharmaceutical Betel Shisanwei Ingredients Pil on the hypothalamic-pituitary-adrenal axis negative feedback function in the chronic depressed rats, and to explore anti-depression mechanisms of Mongolian Pharmaceutical Betel Shisanwei ingredients pil. METHODS: Eighty male Wistar rats were randomly divided into ten groups according to the sugar consumption test (with eight rats in each group): normal control group, model group, fluoxetine group, high-, medium- and low-dose Betel Shisanwei Ingredients Pil groups, RU486 group, high-, medium- and low-dose Betel Shisanwei Ingredients Pil plus RU486 groups. Except normal control group, the other groups were treated with the chronic unpredictable mild stress stimulation combined with lonely rising, to establish depression models. In the meantime, rats of the high-, medium- and low-dose Betel Shisanwei Ingredients Pil groups were given oral gavage of Betel Shisanwei Ingredients Pil (0.2, 0.4, 0.8 g/kg) for 28 days; rats of the normal control group and model group were intragstricaly administered with sodium carboxymethyl celulose; rats of RU486 group were given abdominal subcutaneous injection of RU486 from day 21 after modeling; rats of the high-, medium- and low-dose Betel Shisanwei Ingredients Pil plus RU486 groups were intragstricaly administered with Betel Shisanwei Ingredients Pil (0.2, 0.4, 0.8 g/kg) and subcutaneous injection of RU486 from day 21. RESULTS AND CONCLUSION:Compared with normal control group, cortisone content increased significantly (P < 0.05), the expression of glucocorticoid receptor mRNA in hippocampus, hypothalamus and pituitary gland decreased significantly, and hypothalamic corticotrophin releasing hormone mRNA expression increased significantly in the model group and RU486 group. Compared with model group, cortisone content decreased, the expression of glucocorticoid receptor mRNA in hippocampus, hypothalamus and pituitary gland increased significantly, and hypothalamic corticotrophin releasing hormone mRNA expression decreased significantly in rats treated with Betel Shisanwei Ingredients Pil. Compared with RU486 group, Betel Shisanwei Ingredients Pil administration led to changed in cortisone content, glucocorticoid receptor mRNA expression in hippocampus, hypothalamus and pituitary gland, as wel as hypothalamic corticotrophin releasing hormone mRNA expression. Experimental findings indicate that, Betel Shisanwei Ingredients Pil can directly regulate excessive secretion of glucocorticoid, and improve the dysfunction of hypothalamic-pituitary-adrenal axis central negative feedback through increasing glucocorticoid receptor mRNA expression and decreasing corticotropin releasing hormone mRNA expression. After the hypothalamic-pituitary-adrenal axis negative feedback pathway is blocked, the effect of Betel Shisanwei Ingredients Pil is weakened.

The renal toxicity of rats after a single dose ofMeng-Gen-Wu-Su (mercury) processed products,Meng-Gen-Wu-Su (mercury)-18-composition pill, mercuric sulfide, mercuric chloride, and mercurous chloride was studied. Fifty-four male Wistar rats were randomly divided into nine groups according to body weights (6 rats in each group): normal control group, low and high dose groups (0.033, 0.33 g·kg-1·d-1) ofMeng-Gen-Wu-Su (mercury) processed products, low and high dose groups (0.29, 2.9 g·kg-1·d-1) ofMeng-Gen-Wu-Su (mercury)-18-composition pill, simplified prescription ofMeng-Gen-Wu-Su (mercury)-18-composition pill group (0.26 g·kg-1·d-1), mercuric sulfide group (17.39 mg·kg-1·d-1), mercuric chloride group (4.06 mg·kg-1·d-1) and mercurous chloride group (35.3 mg·kg-1·d-1). After acclimation for one week, once oral administration was given to each group of rats. After 24 h, function and morphological changes of liver and kidney were detected. Mercury accumulation in kidney was determined by inductively coupled plasma optical emission spectroscopy (ICP-OES) and inductively coupled plasma source mass spectrometer (ICP-MS). Apoptosis of renal cell was determined by terminal-deoxynucleoitidyl transferase mediated Nick End Labeling (TUNEL). Renal typeⅢ collagen protein's expression was determined by immunohistochemical (HIC) method and expression changes of MT-1, MT-2 mRNA in kidney were also determined by real-time fluorescence quantitative PCR (real-time-PCR). There was no significant difference of ALT, AST in serum between normal control group and other groups (P>0.05). CREA and UREA in mercurous chloride group were apparently higher than normal control group and low dose group of Meng-Gen-Wu-Su processed products (P<0.01). Hepatic and renal pathologic examination results showed that liver cell of low dose groups ofMeng-Gen-Wu-Su processed products andMeng-Gen-Wu-Su-18-composition pill swelled to a low degree and glomerular disease was not obvious. In high-dose groups ofMeng-Gen-Wu-Su processed products,Meng-Gen-Wu-Su-18-composition pill and mercuric sulfide group, liver and kidney appeared some pathological changes and such changes were more significant in mercuric chloride and mercurous chloride groups. Compared with normal control group and low dose group ofMeng-Gen-Wu-Su processed products, the mercury kidney volume in mercuric chloride and mercurous chloride groups increased significantly (P<0.01). The apoptosis rate of renal cell and expression of typeⅢ collagen protein increased significantly in the groups of mercuric sulfide, mercuric chloride and mercurous chloride (P<0.01). MT-1and MT-2 mRNA gene expression rised significantly in the groups of mercuric chloride and mercurous chloride (P<0.05 orP<0.01). In summary, the rats renal toxicity after a single dose ofMeng-Gen-Wu-Su (Mercury) processed products or MongolianMeng-Gen-Wu-Su (Mercury)-18-composition pill were both far less than that of mercuric chloride or mercurous chloride.

Objective To study the inhibitory mechanism of hydroxysafflor yellow A (HSYA)on abnor-mal proliferation of vascular endothelial cells.Methods Co-cultured model in vitro was established, with supernatant fraction of LS180 human colon adenocarcinoma cells tumor cells and ECV304 human umbilical vein endothelial cells.Then mRNA expressions of vascular endothelial growth factors (VEGF) and VEGF receptors (KDR),basic fibroblast growth factor (bFGF)and bFGF receptors(bFGFR),pro-teoglycan related to cell proliferation (HSPG2),p53 and c-myc were detected using real-time fluores-cence quantitative PCR;protein expressions of VEFG,KDR,bFGF and bFGFR in supernatant fraction were measured by ELISA.Results In the cell co-cultured model,the HSYA concentration of 0.66 and 0.33 mg/L inhibited mRNA and protein expressions of VEGF,KDR,bFGF and bFGFR ,inhibited mR-NA expressions of c-myc and HSPG2,while upregulated p53 mRNA expression.Conclusion HSYA in-hibited angiogenesis and abnormal proliferation of vascular endothelial cells in co-cultured model by regu-lating expressions of VEGF,VEGFR,bFGF,bFGFR,HSPG2,c-myc,wild type p53.HSYA may be a potential tumor angiogenesis inhibitor.

Objective To study the effects of Peiyuan Jieyu Formula ( PYJYF) on rats treated with glu-cocorticoid, its influences on metabolism of the TRP-KYN pathway and the impacts on hippocampal and cortical neuronal plasticity. Methods 70 rats were randomly divided into the following 7 groups ( n=10 in each group): normal blank group, model group, fluoxetine group, mid- and high-dose Tiansi Yin ( TSY) groups, Sini San ( SNS) groups, and PYJYF groups. Rat model of depression was established by intraperitoneal injection of glucocorticoid for 28 days. Sucrose solution consumption and Open-field test were conducted. The content of TRP, KYN, KA and Q in serum was measuredby using HPLC-MS/MS technique. The mRNA expressions of NR and BDNF in brain tissue were measured by using RT-PCR. Results 28 days after the injection of glucocorticoid in model group, the consumption of sucrose solution and total score in open-field test was significantly decreased ( P <0 . 05 ) . The ratios of KYN/TRP and KYN/KA increased significantly ( P<0 . 05 ) . The mRNA expression level of BDNF in hippocampal de-creased significantly ( P<0 . 05 ) . The mRNA expression level of NR increased significantly ( P<0 . 05 ) . Peiyuan Jieyu Formula increased the consumption of sucrose solution (P<0. 05),the total score in open-field test,the ratios of KYN/Q and the mRNA expression level of BDNF in hippocampus(P<0. 05) when compared with the model group. The ratios of KYN/TRP and KYN/KA(P<0. 05) and the mRNA ex-pression level of NR in cortex ( P <0. 05 ) of Peiyuan Jieyu Formula group decreased. Conclusion Peiyuan Jieyu Formula can reduce the metabolism of TRP-KYN, inhibit neurotoxicity, improve neural protection, and promote the hippocampal and cortical neural plasticity.

ObjectiveTo explore the improvement effect of Flos Puerariae， Hoveniae Semen， and their compatibility on acute alcoholic gastric mucosal injury， and lay a foundation for further development of Flos Puerariae， Hoveniae Semen， and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury. MethodThe acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor （15 mL·kg^-1）. A total of 120 male ICR mice were randomly divided into 8 groups， namely， the blank group， model group， omeprazole group （0.026 g·kg^-1）， Flos Puerariae-Hoveniae Semen （compatibility） high， medium， and low-dose groups （29.2，14.6， 7.3 g·kg^-1）， Flos Puerariae group （19.5 g·kg^-1）， and Hoveniae Semen group （19.5 g·kg^-1）， with 15 mice in each group. After one week of adaptive feeding， the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg^-1 for 3 d. From the 4^th day， after 1 h of administration， Erguotou liquid was administrated at the rate of 15 mL·kg^-1 and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration， the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph， and the activity of ethanol dehydrogenase （ADH） in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of nuclear transcription factor-κB （NF-κB） p65 and NF-κB inhibitory protein α （IκBα） were detected by real-time polymerase chain reaction （Real-time PCR）. ResultAs compared with the normal group， the content of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） in serum of mice in the model group was increased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues was increased （P<0.01）， and the mRNA expression of IκBα was decreased （P<0.01）. As compared with the model group， the drunkenness time of the omeprazole group， high and medium-dose compatibility groups， and Flos Puerariae group was prolonged （P<0.05）， the sober up time of the high and medium-dose compatibility groups was shortened （P<0.05）， the ethanol concentration in the serum of the high-dose compatibility group was decreased （P<0.05）， the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased （P<0.05）， the macroscopic injury score of the high， medium， and low-dose compatibility groups and Flos Puerariae group was decreased （P<0.05）， the score of pathological injury in the omeprazole group， high， medium， and low-dose compatibility groups， and Flos Puerariae group was decreased （P<0.01）， the expression of IL-6 in serum of all drug groups was decreased （P<0.05）， the expression of IL-1β in serum of the omeprazole group， high， medium， and low-dose Flos Puerariae groups， and Hoveniae Semen group was decreased （P<0.05）， the expression of TNF-α in serum of high and medium-dose groups was decreased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues of all drug groups was decreased （P<0.05）， and the mRNA expression of IκBα in gastric mucosa tissues of the omeprazole group and high， medium， and low-dose compatibility groups was increased （P<0.05）. As compared with the high-dose compatibility group， the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened （P<0.01）， the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged （P<0.01）， the concentration of ethanol in the serum of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group increased （P<0.05）， the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased （P<0.05）， the pathological injury score of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， the content of IL-1β in serum of low-dose compatibility group， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， and the mRNA expression of IκBα in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased （P<0.05）. As compared with the medium-dose compatibility group， the drunkenness time in the Hoveniae Semen group was shortened （P<0.05）， the sober up time in the Flos Puerariae group was prolonged （P<0.05）， the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased （P<0.01）， and the content of IL-1β in serum of the low-dose compatibility group， the Flos Puerariae group， and Hoveniae Semen group was increased （P<0.05）. As compared with the low-dose compatibility group， the pathological injury score of the Hoveniae Semen group was increased （P<0.05）. ConclusionFlos Puerariae， Hoveniae Semen， and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice， which may be related to the inhibition of the expression of NF-κB signal pathway in gastric mucosa， and the high-dose compatibility group has the optimal effect.

ObjectiveTo observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang （HYT） on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020. MethodA total of 128 male Wistar rats were randomly divided into a blank group， a model group， a euthyrox group （20 μg·kg^-1）， a HYT group （12.06 g·kg^-1）， a HYT without Sargassum （HYT-H） group （9.90 g·kg^-1）， a HYT without Glycyrrhizae Radix et Rhizoma （HYT-G） group （10.26 g·kg^-1）， a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma （HYT-HG） group （8.10 g·kg^-1）， and a Sargassum and Glycyrrhizae Radix et Rhizoma （HG） group （3.96 g·kg^-1）. The blank group was given deionized water by gavage， and the others were given propylthiouracil （PTU） to replicate the goiter pathological model. Euthyrox was taken as a positive control drug， and the rest of the Chinese medicine groups were given the corresponding decoction by gavage， the material was collected 12 hours after the last dose. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， alkaline phosphatase （ALP）， and the contents of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， malondialdehyde （MDA）， reactive oxygen species （ROS） in liver tissue were detected in each group. The pathological changes in the liver were observed via hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1 （Keap1）， nuclear factor E₂-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， p53 and Caspase-3 in liver tissues. Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways. ResultCompared with control group， the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues （P<0.05， P<0.01）， significantly reduced activities of SOD and GSH-Px in the liver （P<0.01）， significantly increased mRNA expression of Keap1 （P<0.01）， and significantly decreased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. Compared with the model group， the HYT group manifested significantly reduced serum levels of AST， ALT， and ALP （P<0.05， P<0.01）， significantly reduced contents of MDA and ROS in liver tissue （P<0.01）， significantly increased the activities of SOD and GSH-Px （P<0.01）， significantly decreased mRNA expressions of Keap1， p53， and Caspase-3 （P<0.01）， and significantly increased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. ConclusionUnder the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020， Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the HYT on oxidative stress in the liver of goiter rats had different effects. The HYT that contains Sargassum and Glycyrrhizae Radix et Rhizoma has a protective effect on the liver of goiter rats， and the effect is better than that of the HG group， the euthyrox group， and the incomplete groups. Its mechanism may be related to activating the Nrf2/HO-1 signaling pathway to alleviate liver oxidative stress and inhibiting the p53/Caspase-3 signaling pathway to reduce hepatocyte apoptosis.