Synergistic nourishing 'Yin' effect of iridoid and phenylpropanoid glycosides from Radix Scrophulariae in vivo and in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Radix Scrophulariae (RS), is a renowned traditional Chinese medicine used as nourishing 'Yin'. The iridoid glycosides (IG) and phenylpropanoid glycosides (PG) are main chemical constituents in RS. However, there had been no pharmacological experiment studies of synergy between IG and PG. Due to the constituents interactions, exploring their synergy profile is of great important for explaining the essence of nourishing 'Yin' efficacy of RS. AIM OF STUDY: The present study was undertaken to evaluate synergistic nourishing 'Yin' effect of IG and PG from RS in vivo and in vitro through their immunoregulation and antioxidant activities. MATERIALS AND METHODS: In this study, IG and PG fractions in RS were isolated and identified by High Performance Liquid Chromatography coupled with tandem quadrupole time-of-flight Mass Spectrometry (HPLC-Q-TOF-MS). The synergistic nourishing 'Yin' effect of two fractions were investigated in vivo and in vitro with thyroxine-induced 'Yin' deficiency (YD) mice model and primary splenic lymphocyte, respectively. The exterior syndrome signs and serologic and cellular biomarkers changes were detected. Then, the synergistic coefficient (SC) of IG and PG on every pharmacodynamics index were calculated by Webb method. RESULTS: Compared with model and mono-therapy group (IG or PG group), IG combined with PG group significantly ameliorated YD by exerting immunoregulation and antioxidant effects. Based on the SC, IG and PG possessed a synergistic effect on heart rate, average speed, upright times, spleen index, LPO, SOD, IL-6, Na+-K+-ATP enzyme in vivo, and cAMP/cGMP, IFN-γ/IL-10, and MDA in vitro with SC > 1. CONCLUSIONS: The nourishing 'Yin' benefits were clearly produced when IG and PG were used in combination, which provided the scientific evidence of multiple-components and multiple-approach synergistic effect of Chinese traditional herbal medicine to control and management of diseases.

The Chinese herbal formula Zhibai Dihuang Granule treat Yin-deficiency-heat syndrome rats by regulating the immune responses.

ETHNOPHARMACOLOGICAL RELEVANCE: Zhibai Dihuang Granule (ZDG), a traditional Chinese medicine (TCM) made from eight Chinese herbs, has been classically used to treat Yin-deficiency-heat (YDH) syndrome. ZDG is well known with the therapeutic efficacy of nourishing Yin and decreasing internal heat in clinic, but the mechanism of ZDG's therapeutic effect is still not clear. MATERIALS AND METHODS: High doses of triiodothyronine (T3) were given intraperitoneally to induce Hyperthyroid YDH syndrome in SD rats. The animals were then treated with ZDG for one week. The iTRAQ-coupled with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) technique was used to screen the differentially expressed serum proteins between ZDG treated rats and YDH syndrome rats. The differentially expressed proteins were analyzed by bioinformatics method and were verified by enzyme-linked immunosorbent assay (ELISA). RESULTS: A total of 55 differentially expressed proteins were identified, including 23 up-regulated proteins (>1.25 fold, p < 0.05) and 32 down-regulated proteins (<0.80 fold, p < 0.05). Among the differentially expressed proteins, 26 proteins returned to normal after ZDG treatment. Bioinformatics analysis showed that these proteins were mainly involved in immune response, including regulation of immune system process, complement activation, and humoral immune response mediated by circulating immunoglobulin. ELISA revealed significantly increased levels of Zinc-alpha-2-glycoprotein (Azgp1), L-selectin, C-reactive protein (Crp), Plasminogen (Plg), Kininogen 1 (Kng1), and significantly decreased levels of Mannose binding lectin 2 (Mbl2) and Complement C1qb chain (C1qb) in ZDG treated rats compared with YDH syndrome rats. Bioinformatics analyses indicated that Azgp1 participated in antigen processing and presentation, Crp, C1qb, and Mbl2 were involved in complement activation, while L-selectin, Plg, and Kng1 were involved in regulating the inflammatory response. CONCLUSIONS: Our study provides experimental evidence to understand the therapeutic mechanism of ZDG in YDH syndrome. The results suggested that ZDG may regulate the complement activation and inflammatory response, and promote the ability to recognize antigens to alleviate YDH syndrome.

[Nourishing-yin effect and mechanism of different parts of Cornu Elaphuri Davidiani in rats].

OBJECTIVE: To investigate nourishing-yin effect and mechanism of different parts of Cornu Elaphuri Davidiani in rats. METHOD: The model of yin asthenia rats was built by thy roxine. The substance metabolism, pain threshold, hormone levels and biochemical indicators in serum were measured. RESULTS: The ethanol extract of Cornu Elaphuri Davidiani could regulate the substance metabolism and raise the pain threshold in yin asthenia model rats. Furthermore, it could regulate the hormone levels, biochemical indicators in serum and it could improvte the antioxidant ability. CONCLUSION: The ethanol extract of Cornu Elaphuri Davidiani showed significant nourishing-yin effect in rats and the possible mechanism is correlated with regulating the neuroendocrine network.

[Mathematical analysis of characteristics of glucocorticoid-induced yang deficiency or yin deficiency syndrome in animal models based on information entropy theory].

OBJECTIVE: Based on information entropy theory, this study analyzes the experimental indicators of glucocorticoid-induced traditional Chinese medicine yang deficiency or yin deficiency syndrome in animal models, thus laying the foundation of defining and interpreting the model state. METHODS: Data of biochemical indicators from 24 groups of animal models with different modeling conditions (type of modeling agent, dosage of modeling agent and modeling time) were collected. Information gain values for three study objects (type of modeling agent, dosage of modeling agent and modeling time) were calculated respectively after standardization, and then characteristics of yang deficiency or yin deficiency syndrome models were interpreted with these values and ranking map. RESULTS: Greatest information gain values of most indicators were got when the study object was the type of modeling agent, which is the most important factor in the differentiation of model state. With this study object, triglyceride (TG), total cholesterol (TC), total protein (TP), albumin (ALB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and corticosteroid (CS) got larger information gain values than testosterone (T), lactic dehydrogenase (LDH), and cyclic adenosine monophosphate (cAMP). This indicated that the former seven indicators may be significantly different between the two animal models induced by hydrocortisone and dexamethasone, respectively. In the study of the modeling time, AST may be affected more than others. In the study of the dosage of modeling agent, TC and ALB may be affected more in the model of hydrocortisone and no indicators were significantly affected by the dosage of modeling agent in the dexamethasone model. CONCLUSION: Mathematical method based on information entropy theory allows researchers to analyze experimental data in several experiments at the same time and to interpret the trend and stability of indicators. Based on this study and by analyzing the impacts of different modeling conditions on the indicators, elucidations of the characteristics of glucocorticoid-induced yang deficiency or yin deficiency syndrome in animal models and some biochemical indicators for model comparison are in need.

[Effects of Sisheng Decoction on spontaneous activity and serum concentration of malondialdehyde in mice with yin deficiency syndrome].

OBJECTIVE: To assess the therapeutic effects of Sisheng Decoction, a compound traditional Chinese herbal medicine, on a mouse model of yin deficiency syndrome induced by thyroid hormone, and to make the preliminary study on its mechanisms. METHODS: Simultaneous modeling and treatment were carried out. Sixty mice were randomly divided into six groups: normal group, yin deficiency model group, low-, medium- and high-dose Sisheng Decoction group and Shengmai oral liquid group. Normal group and yin deficiency model group were administered with double distilled water. Spontaneous activity and serum concentration of malondialdehyde in different groups were detected. RESULTS: The symptoms of yin deficiency syndrome such as xerostomia, dysphoria and fervescence were improved in the Sisheng Decoction groups. Compared with the yin deficiency model group, the spontaneous activity was increased and the serum concentration of malondialdehyde was decreased in the Sisheng Decoction groups (P<0.05). There was no significant difference between the Sisheng Decoction groups and the Shengmai oral liquid group (P<0.05). CONCLUSION: Medium- or high-dose Sisheng Decoction is effective for nourishing yin, clearing heat, engendering liquid and allaying thirst. The above effects of Sisheng Decoction may be realised by improving the spontaneous activity and resisting oxidative damage.

[Effects of Semen Ziziphi Spinosae on the anxiety behavior of the yin deficiency mice].

OBJECTIVE: To observe the effects of Semen Ziziphi Spinosae on the anxiety behavior of the yin deficiency mice, to filter the active material of Semen Ziziphi Spinosae and explore its anti-anxiety mechanism. METHODS: The yin deficiency mice model with intragastrical 320 mg/kg thyroid tablet for nine days exception normal control group. To observe yin deficiency model mice anxiety action by means of the elevated plus-maze and light-dark transitions of mice. After the experiment, peeled off cerebra at once. The left saved in liquid nitrogen for assaying GABA and Glu with TLCS, and right saved in formalin for the protein expression of GABAaR1 and NMDAR1 with immunohistochemical method. RESULTS: (1) Compared with yin deficiency model group, Tabellae Diazepami group and the middle alcohol extracts group increased instinctively in the percentage of time and degree entering open-arm in eleveated plus maze, and the passing times in light-dark transitions increased instinctively (P<0.05). (2) Compared with yin deficiency model group, the contents of GABA increased evidently (P<0.05) and Glu reduced evidently (P<0.05) in Tabellae Diazepami group and the middle alcohol extracts group. (3) Compared with yin deficiency model group,the expression of GABAAR1 increased evidently (P<0.05) and NMDAR1 reduced evidently (P<0.05) in the diszepam group and the middle alcohol extracts group. CONCLUSIONS: The alcohol extracts of Semen Ziziphi Spinosae have the anti-anxiety effects instinctively. Its mechanism may be related to increasing the GABA and expression of GABAAR1 and reducing the Glu and expression of NMDAR1.

[Thyroid-induced hyperthyroidism Yin-deficiency model of rats].

OBJECTIVE: To establish a rat model of hyperthyroidism Yin-deficiency. METHODS: SD rats separate male and female were randomly divided into four groups, a total of eight groups, each with six. The first group for the control group, 2 to 4 for the experimental drug-treated group, including thyroid-low, medium and high-dose group. In addition to the first group lavage volume of saline, the other groups were given the concentration of 2. 5, 5, 10 mg/kg-saline suspension of the thyroid. Once a day, 30 days for delivery. RESULTS: Compared with the control group, with the exception of Group 4 female rats serum T3, T4, FT3, FT4 values were significantly higher (P < 0.01), serum TSH was significantly lower (P < 0.05), the other treatment groups had different values no significant difference (P > 0.05). Group 4 that female rats modeling was success. CONCLUSION: Administration of thyroid-Suspension 10 mg/ kg for 30 d, female rats hyperthyroidism Yin-deficiency rat model is success.

[Effects of herbs for replenishing yin to tonify kidney on testicular ultrastructural changes in rats with glucocorticoid-induced kidney deficiency].

OBJECTIVE: To investigate the effects of herbs for replenishing yin to tonify kidney on testicular ultrastructural changes in rats with glucocorticoid-induced kidney deficiency. METHODS: Thirty rats were randomized into three groups: normal control group, untreated group and Erdi Biejia Decoction (EDBJD)-treated group. Kidney deficiency rat model was induced by glucocorticoid. Testicular structure and ultrastructure were observed under the light microscope and the transmission electron microscope. RESULTS: The light microscope revealed the decreased thickness of seminiferous epithelium layer, reduced number of spermatogenic cells, interstitial tissue edema, and a qualitative reduction in the number of interstitial cells and hematoxylin-eosin stained cytoplasm in the untreated group. The transmission electron microscope revealed loose structure of seminiferous epithelium, degenerative mitochondria, and increased amounts of lysosome, presence of numerous cytoplasmic vacuolation and chromatin in interstitial cell cytoplasm in the untreated group. The aggregates of heterochromatin around the nuclear membrane and the shrinkage of nucleus were also observed. Compared to the untreated group, these pathological changes were inconspicuous in EDBJD-treated group. CONCLUSION: EDBJD as replenishing yin to tonify kidney herbs can suppress glucocorticoid induced pathological changes in the testes of rats.

[Intervention of maiwei dihuang oral liquid on hormonotherapy in treating active systemic lupus erythematosus].

OBJECTIVE: To observe the intervention of Maiwei Dihuang Oral Liquid (MDOL) on hormonotherapy in treating active systemic lupus erythematosus (SLE). METHODS: Sixty SLE patients in active stage were randomly and equally allocated into two groups, and administered with prednisone, which was medicated in initial dose of 0.5-1 mg/kg, and with the dose being reduced conditionally 6-8 weeks. To the 30 patients in the treated group 10 ml MDOL twice daily was given additionally. The therapeutic course was 3 months. RESULTS: The therapeutic effect in the treated group was better than that in the control group (P < 0.05). Systemic lupus erythematosus disease activity index (SLEDAI) was significantly improved in both groups (P < 0.01), but was superior in the treated group (P < 0.05). The dose of prednisone used was significantly reduced (P < 0.01), and the scores of Yin-deficiency fire-flourishing syndrome were obviously decreased (P < 0.01) in the treated group while in the control group, these indexes were unchanged (P > 0.05), the difference between the two groups was significant (P < 0.01). The occurrence of adverse reaction was significantly lower in the treated group than that in the control group (P < 0.05). CONCLUSION: MDOL can obviously improve the effect of hormonotherapy in SLE, it has advantages in reducing the dosage used and antagonizing the adverse reactions of glucocorticoid.

[Clinical study on intervention of liuwei dihuang pill on hormonotherapy in treating nephrotic syndrome].

OBJECTIVE: To observe the intervention of liuwei dihuang pill (LDP) on therapeutic effectiveness and adverse reaction of hormonotherapy in treating nephrotic syndrome. METHODS: Patients allocated in two groups were medicated with initial dose of prednisone 1 mg/kg once a day at 8 am in the morning. After being medicated for 8 to 12 weeks, the dose of prednisone was decreased by 5.0 mg every 2 weeks till 0.5 mg/kg per day. Then the medication was changed to that two days' dosage orally take once a day with the daily dose reduced by 5.0 mg/kg every 2 to 3 weeks, and maintained at 0.4 mg/kg once every two days. At same time, necessary symptomatic treatment was given. To the treated group oral administration of LDP 8 capsules was given additionally, 3 times per day until prednisone decreased to maintenance dose. RESULTS: Therapeutic effect in the treated group was significant better than that in the control group (P < 0.05). Urinary protein, plasma albumin, triglyceride (TG) and total cholesterol (TC) in both groups were obviously improved (P < 0.05 or P < 0.01). However, as compared with the control group, the improvement was better, and the recurrent rate was lower (P < 0.05) in the treated group. Scores of Yin-deficiency caused excessive Fire syndrome and incidence rate of adverse reaction in the treated group were lower than those in the control group (P < 0.05 or P < 0.01). CONCLUSION: LDP can markedly improve the therapeutic effectiveness and counteract the adverse reaction of hormonotherapy in treating nephrotic syndrome, and reduce the recurrence of the disease.

[Effects of di-zhen granules on catecholamine transmitter and gonadotropin of clamacteric rats with deficiency of yin brings about the interior heat-syndrome].

Fifteen-month-old Sprague-Dawley rats were given with some kinds of hot property of herb to produce model of Deficiency of yin brings about the interior heat-syndrome. The weight, water drinking volume, and body temperature of model rats were observed at the process of producing model and the treatment of di-zhen granules. Hypothalamic NE, DA and plasma NE, E concentration were determined by high-performance liquid chromatography, and serum E2, FSH, LH levels were determined by radioimmunoassay. The results were that after having been given the hot property of herbs for 2 weeks, the model rats increased water drinking volume and body temperature, which is similar to the appearance of patients with Deficiency of yin brings about the interior heat syndrome, and after having been treated with di-zhen granules, the water drinking volume and temperature of model rats decreased significantly (P < 0.01). The hypothalamic NE, DA, plasma NE, E, and the serum FSH, LH of the group treated with di-zhen granules were significantly lower than that of the model group and the normal group(P < 0.01). The results show that our model mentioned above possesses some characteristic features of the human climacteric syndrome which exhibits Deficiency of yin brings about the interior heat-syndrome. Di-zhen granules may regulate the function of GnRH neurons and other neurotic nucleus in hypothalamus, and then rectify the endocrine disorders of climacteric.

[Effect of taibai-ginseng on the mice pathology model of "kidney yindeficiency" induced by excessive throxine].

Effect of Taibai-ginseng on mice pathology model of "kidney yindeficiency" induced by excessive throxine was observed in this paper. The result showed taibai-ginseng could improve the pathology symptoms remarkably. It could restrain the rise of body oxygen consuming, body temperature, capacity of drinking water and the decline of pain threshold.

[Experimental studies of Chinese taihe chicken on effects of "Qi" reinforcing and "Yin" nourishing].

The "Qi" deficiency and "Yin" deficiency rats were fed with Chinese Taihe Chicken or common Chicken. The results showed: Chinese Taihe Chicken and common Chicken could all improve in part the "Qi" deficiency of rats caused by forced swimming, furthermore, the effect of Chinese Taihe Chicken was better than that of common Chicken. Chinese Taihe Chicken could obviously reduce the activity of adenosine triphosphatase of kidney tissue of "Yin" deficiency rats caused by injecting thyroxine. It suggested that Chinese Taihe Chicken could nourish the "Yin".

[Effect of zuogui wan on the contents of hypothalamic monoaminic transmitters and body weight in rats treated with monosodium glutamate neonatally].

OBJECTIVE: To evaluate the interrelationship between the traditional Chinese medicine the Kidney-Yin Deficiency Syndrome and the metabolism of hypothalamic monoaminic neurotransmitters after the lesion of arcuate nucleus (ARC). METHODS: Rats were injected with monosodium glutamate (MSG, at 2, 4, 6, 8, 10 days after born. 4 mg/g body weight, s.c.) during the neonatal period, then the contents of hypothalamic monoaminic transmitters were determined with HPLC and the changes of increase of body weight were observed. RESULTS: Decreases in levels of norepinephrine (NE), dopamine (DA) associated with its metabolite, 3,4-dihydroxyphenyl acetic acid (DOPAC) were observed, while vanillylmandelic acid (VMA), metabolite of NE, serotonin (5-HT) and 5-hydroxyindole acetic acid (5-HIAA) were unchanged. Furthermore, adult rats with neonatal MSG treatment also demonstrated growth retardation, obesity but lower body weight, and shorter length of naso-anal. Zuogui Wan, classical drugs of nourishing and recuperating the Kidney-Yin, relieved the disorder of the metabolism of hypothalamic monoaminic transmitter in MSG-rats. CONCLUSION: The metabolic disorders of the central monoaminic transmitters may be related with the Deficiency of the Kidney-Yin.

[Effect of yin and qi tonifying herbs on hyperthyroid model of yin-deficient rats].

The article reports the effect of Yin and Qi tonifying herbs (Radix Rehmanniae, Radix Ophiopogonis and Pseudostellaria Heterophylla) on hyperthyroid model of Yin-deficient rats. The model rats were made Yin-deficient by injecting thyroxine. Observation showed that after injection, indexes such as anus temperature, oxygen consumption, weight, diet, activity, general conditions, triiodothyronine (T3) and thyroxine (T4) in serum were markedly changed. It is thus suggested that the Yin and Qi tonifying mixture of the above three herbs can regulate and correct Yin-deficiency in hyperthyroid model of Yin-deficient rats.

[134 patients with adult primary nephrotic syndrome treated with combined traditional Chinese medicine and Western medicine].

This paper studied the method and effect of adult primary nephrotic syndrome (PNS) with the integrated traditional Chinese and Western medicine (TCM-WM) treatment. 134 patients were divided into WM group (66 cases) and TCM-WM group (68 cases). Both groups were mainly treated with corticosteroid. The latter group was treated with corticosteroid plus Chinese medicinal herbs. The results showed that total effective rate of WM group and TCM-WM group was 56.1% and 85.3% respectively, while the occurrence rate of side effect was 48% and 14.8% respectively. It indicated that corticosteroid plus Chinese medicinal herbs could enhance curative effect and reduce side effect.

[Herbal decoction of qingwen baidu yin in treating endotoxic fever in rabbits].

Qingwen Baidu Yin (QBY) has good curative effects on the endotoxic fever of rabbits induced by injecting endotoxin of E. Coli. The test group was given QBY orally, while the control group was given NS orally instead. Result showed QBY could: (1) Markedly inhibit the fever, it was effective in reducing febrile curve. delta T and TRI5 of the test group were smaller (P < 0.001). (2) Ameliorate the leukocytopenia and leukocytosis, and improve thrombocytopenia. (3) Antagonize hyperviscosity syndrome and had the actions of depolymerization and dilution. (4) In test group, the increased cAMP content in plasma was reduced, and the decreased cGMP content raised, the ratio of cAMP and cGMP was nearly normal. All these provided the clue in elucidating the essence of "Excessive Yang causes Heat" and "Predominance of Yang leads to disorder of Yin". (5) Pathomorphological examination showed that QBY had the functions of protecting the internal organs and reducing the organic damage induced by endotoxing in rabbits.