ABSTRACT
Based on the metabolomics, this paper systematically analyzed the metabolic substance basis of Zuogui Pills and Yougui Pills in syndrome differentiation and treatment of diminished ovarian reserve(DOR), so as to provide a scientific basis for the traditional Chinese medicine(TCM) syndrome differentiation and treatment of DOR. Patients with DOR of kidney-Yin deficiency syndrome were collected from outpatient department of hospitals and treated with Zuogui Pills for 12 weeks. And kidney-Yang deficiency syndrome were treated with Yougui Pills for 12 weeks. Based on the non-targeted metabolomic research techniques, the potential biomarkers of Zuogui Pills and Yougui Pills in the treatment of DOR with kidney-Yin deficiency and kidney-Yang deficiency, respectively, were screened out, and metabolic pathways of biomarkers were analyzed. The pregnancy rate, basic serum hormone levels [basal follicle-stimulating hormone(bFSH), basal-luteinizing hormone(bLH), basal-estradiol(bE_2), and anti-Müllerian hormone(AMH)], TCM syndrome type score, and Kupperman score were recorded and statistically analyzed after treatment. The results showed that 23 patients with DOR of kidney-Yin deficiency syndrome and 25 patients of kidney-Yang deficiency syndrome were collected. Twenty-six differential metabolites, including L-carnitine, acetyl-CoA, coenzyme A, and coenzyme Q_(10)(CoQ10), were mapped to 12 metabolic pathways in patients with kidney-Yin deficiency treated with Zuogui Pills. Twenty-two differential metabolites, such as adipoyl-CoA, L-lysine, lysine arginine, and α-tocopherol, were mapped to 11 metabolic pathways in patients with kidney-Yang deficiency. After treatment, bFSH and bLH of patients with DOR were significantly lower than those before treatment(P<0.05). Although the comparison of bE_2 and AMH had no significant differences, there was a improvement trend. The TCM syndrome type score and Kupperman score of patients with DOR after TCM treatment were significantly lower than those before treatment(P<0.05).
ABSTRACT
OBJECTIVE：To investigate the contents of 5 kinds of heavy metal as copper （Cu），arsenic（As），cadmium （Cd），mercury（Hg）and lead （Pb）in Yougui pills ，and to evaluate its safety risk. METHODS ：Using yttrium （89Y），indium （115In）and bismuth （209bi）as internal standard ，the contents of each element were determined by ICP-MS. ICP-MS condition included that atomization gas flow rate was 0.95 L/min，auxiliary gas flow rate was 1.2 L/min，plasma gas （argon）flow rate was 18 L/min，pump speed was 30 r/min. RF power of inductively coupled plasma was 1 200 W，the voltage in simulation stage was 1 750 V，the voltage in pulse stage was 1 300 V，the voltage of deflection device was －12 V，and the detector was in analog and pulse dual-mode. The determination methods of various elements were investigated ，and 45 batches of marketed Yougui pills were determined. Hazard index （HI）was used to analyze the non-carcinogenic risk of each element and calculate the maximum residual limit（MRL）of each element. RESULTS ：The linear range of Cu ，As，Cd，Hg and Pb ranged from 10-200，1-50，0.4-30，0.2-6 and 2-100 μg/L（all r＞0.999 0），respectively. The limits of quantitation were 0.67，0.23，0.20，0.07，0.27 μg/L. The limits of detection were 0.20，0.07，0.06，0.02，0.08 μg/L. RSDs of precision，stability and reproducibility tests were all lower than 3.5％ （n＝6 or n＝5）. Average recoveries were 92.96％-100.89％（RSD＝2.23％-3.62％，n＝3）. Average contents of Cu ，As，Cd，Hg and Pb in 45 batches of Yougui pills were 2.72，0.28，0.07，0.05，0.62 mg/kg，and superimposed HI of each element was less than 1. The contents of 5 kinds of heavy metals in Yougui pills were lower than the proposed MRL （MRL of Cu ，As，Cd，Hg and Pb were 20，2，1，0.2，5 mg/kg or 111.11，4.44，2.22，1.48，8.89 mg/kg respectively ）. CONCLUSIONS ：Established method can be used for the determination of content of 5 kinds of heavy metal in Yougui pills ；the heavy metal pollution rate of marketed Yougui pills is low and the safety risk is small.
ABSTRACT
AIM To observe the effects of Zuogui Pills and Yougui Pills on the proliferation of BMSCs and the expression of apoptosis after osteogenic and adipogenic differentiation in ovariectomized rats.METHODS Of sixty female SD rats,forty rats followed by bilateral ovariectomy were randomly divided into ovariectomy group (1.0 g/kg distilled water),Zuogui Pills group (9.45 g/kg Zuogui Pills) and Yougui Pills group (10.26 g/kg Yougui Pills) and Bujiale group (0.09 mg/kg estradiol valerate),another ten rats as control group (1.0 g/kg distilled water),ten rats bilateral excision of a small amount of fat around the ovary was treated as sham operation group (1.0 g/kg distilled water).After 12 weeks of administration,the rats were killed,BMSCs were cultured in vitro.Cell proliferation was detected by MTT assay.Western blot was used to detect the protein expressions of Caspase-3 and Bcl-2.RE-SULTS MTT assay showed that the proliferation of BMSCs were promoted in Zuogui and Yougui Pills groups,and the proliferation effect in Zuogui Pills group was better than that in Yougui Pills group.After osteogenic differentiation,as compared with the control group,the Caspase-3 expression of ovariectomy group was up-regulated (P ＜0.05) and the expression of Bcl-2 was down-regulated (P ＜0.01).As compared with ovariectomy group,the expression of Caspase-3 was decreased and the expression of Bcl-2 was up-regulated in Zuogui Pills group and Yougui Pills group.After adipogenic differentiation,as compared with the control group,the expressions of Caspase-3 were significantly up-regulated and Bcl-2 were down-regulated in the ovariectomy group,Zuogui Pills group and Yougui Pills group (P ＜ 0.05).After osteogenic and adipogenic differentiation Zuogui Pills group and Yougui Pills group were significant different (P ＜ 0.05).CONCLUSION Zuogui Pills and Yougui Pills both can inhibit the apoptosis of BMSCs after osteogenic and adipogenic differentiation in ovariectomized rats.Zuogui Pills can promote BMSCs osteogenesis differentiation while Yougui Pills can promote adipogenic differentiation.
ABSTRACT
AIM To research the antiasthmatic effects of Zuogui Pills,Yougui Pills and Yupingfeng Powder and their mechanism of action.METHODS Male Brown Norway rats were randomly divided into five groups.OVA was used to induce the model of chronic asthma.Five groups of asthmatic rats were given Zuogui Pills,Yougui Pills,Yupingfeng Powder by gavage once daily for 14 d,respectively.The normal and the model group were given normal saline.Penh value,serum Thl,Th2 inflammatory cytokines,and CROT and ACTH protein levels were determined.Pathological changes of airway remodeling were measured.RT-PCR method was used to measure the expression of CRH mRNA in hypothalamus,and TGF-β1 and Smad3 mRNA in lung tissues.RESULTS Penh values of Yupingfeng Powder group and Yougui Pills group were significantly lower than that of Zuogui Pills group.The Thl inflammatory cytokine of Yupingfeng Powder group was significantly higher than that of Zuogui Pills group,while the Th2 cytokine was significantly lower than that of Zuogui Pills group.In Yougui Pills group,only IL-5 was significantly lower than that of Zuogui Pills group.Serum CROT,ACTH protein and CRH mRNA of Yupingfeng Powder group were significantly higher than those in Zuogui Pills group.No significant difference was found between Yupingfeng Powder group and Zuogui Pills group with regard to TGF-β1 and Smad3 mRNA.Smad3 mRNA of Yougui Pills group was significantly higher than that of the model group.Airway remodeling was significantly reduced by Yupingfeng Powder and Zuogui Pills.The number of goblet cells in Yougui Pills group was significantly lower than that in Zuogui Pills group.CONCLUSION All the three formulaes possess significant antiasthma effects.Regulation effects of Yupingfeng Powder on Thl/Th2 balance,HPA and TGF-β1/Smad3 pathway are strong.Compared with Zuogui Pills,Yougui Pills possesses weaker TGF-β1/Smad3 regulative effects and stronger HPA regulative effects.
ABSTRACT
AIM To observe the effects of Zuogui Pills and Yougui Pills on the proliferation of BMSCs and the expression of apoptosis after osteogenic and adipogenic differentiation in ovariectomized rats.METHODS Of sixty female SD rats,forty rats followed by bilateral ovariectomy were randomly divided into ovariectomy group (1.0 g/kg distilled water),Zuogui Pills group (9.45 g/kg Zuogui Pills) and Yougui Pills group (10.26 g/kg Yougui Pills) and Bujiale group (0.09 mg/kg estradiol valerate),another ten rats as control group (1.0 g/kg distilled water),ten rats bilateral excision of a small amount of fat around the ovary was treated as sham operation group (1.0 g/kg distilled water).After 12 weeks of administration,the rats were killed,BMSCs were cultured in vitro.Cell proliferation was detected by MTT assay.Western blot was used to detect the protein expressions of Caspase-3 and Bcl-2.RE-SULTS MTT assay showed that the proliferation of BMSCs were promoted in Zuogui and Yougui Pills groups,and the proliferation effect in Zuogui Pills group was better than that in Yougui Pills group.After osteogenic differentiation,as compared with the control group,the Caspase-3 expression of ovariectomy group was up-regulated (P ＜0.05) and the expression of Bcl-2 was down-regulated (P ＜0.01).As compared with ovariectomy group,the expression of Caspase-3 was decreased and the expression of Bcl-2 was up-regulated in Zuogui Pills group and Yougui Pills group.After adipogenic differentiation,as compared with the control group,the expressions of Caspase-3 were significantly up-regulated and Bcl-2 were down-regulated in the ovariectomy group,Zuogui Pills group and Yougui Pills group (P ＜ 0.05).After osteogenic and adipogenic differentiation Zuogui Pills group and Yougui Pills group were significant different (P ＜ 0.05).CONCLUSION Zuogui Pills and Yougui Pills both can inhibit the apoptosis of BMSCs after osteogenic and adipogenic differentiation in ovariectomized rats.Zuogui Pills can promote BMSCs osteogenesis differentiation while Yougui Pills can promote adipogenic differentiation.
ABSTRACT
Objective To observe the therapeutic effect and security of Thermal Moxibustion combined with Yougui Pills in the treatment of premature ovary failure(POF) of spleen and kidney Yang-deficiency.Methods One hundred and forty-two patients with POF of spleen and kidney Yang-deficiency were randomly divided into the treatment group and control group,71 cases in each group.The observation group were treated with Thermal Moxibustion combined with Yougui Pills,while the control group took oral Yougui Pills,12 weeks as a course of treatment.After treatment,the clinical therapeutic effects,K upperman score,related hormones levels(E2,FSH,LH) and occurrence rate of adverse reactions were compared between the two groups.Results The total efficacy rate of the observation group was 95.77％,which of the control group was 77.46％,and the observation group was significantly better than the control group(P＜0.05);there were significant improvement in the Kupperman score and levels of related hormones after treatment in the two groups(P＜0.05),but the improvement degree of the observation group was superior to the control group(P＜0.05).Conclusion Thermal Moxibustion combined with Yougui Pills can alleviate the clinical symptoms with high safety and higher clinical application value
ABSTRACT
Objective To observe the effects of Yougui Pills (YP) intervention on SREBP pathway related factors of sterol regulatory element binding protein-1c ( SREBP-1c) , cholesteryl ester transfer protein ( CETP) and fatty acid synthase ( FAS) protein and gene expression in kidney yang deficiency hyperlipidemia rat model. Methods Sixty SD rats were divided into normal control group, model group (in the dosage of 2.43 g/kg), and YP group. Rat model of kidney yang deficiency hyperlipidemia was induced with intramuscular injection of hydrocortisone. Normal control group and model group were given normal saline, and YP group was given YP suspension intragastrically. After treatment, the hepatic pathomorphology of rats in the three groups was examined, and the contents of serum lipids were examined. Hepatic SREBP-1c, CETP and FAS protein expression was detected by Western-blotting method, and their mRNA expression was detected by real-time polymerase chain reaction ( RT-PCR) . Results Compared with the normal control group, serum triglyceride ( TG ) , total cholesterol ( TC ) , low-density lipoprotein cholesterol ( LDL-C ) contents were significantly increased (P<0.01), high-density lipoprotein cholesterol (HDL-C) was significantly decreased (P<0.01), and hepatic pathomorphological changes of hyperlipoidemia were obvious in the model group. Compared with the model group, TG, TC, LDL-C contents were significantly decreased ( P<0.01) , and HDL-C was significantly increased (P<0.01) in YP group. Model group had higher SREBP-1c, CETP and FAS mRNA and protein expression than the normal control group ( P<0.05) , while YP group had lower SREBP-1c, CETP and FAS mRNA and protein expression than the model group ( P<0.05) . Conclusion YP can decrease the blood lipid levels probably by down-regulating the expression levels of SREBP-1c, CETP and FAS gene and protein related to the the SREBP pathway in rats of kidney yang deficiency hyperlipidemia induced by intramuscular injection of hydrocortisone.
ABSTRACT
Objective To observe the therapeutic effect of Yougui Pills and its separate formula for the treatment of kidney-yang deficiency syndrome. Methods A total of 60 patients of kidney-yang deficiency syndrome were randomly divided into treatment group and control group, 30 cases in each group. Treatment group was treated with Yougui Pills, and the control group was given the separate formula from Yougui Pills which had the actions of warming kidney yang. Four weeks constituted one treatment course and the treatment covered 2 courses. The changes of major clinical symptom scores, total effectives rate and associated indexes such as peripheral white blood cells, serum antibodies, and complements were compared between the two groups following the treatment for 8 weeks. Results The total effective rate, and improvement of major clinical symptoms and the peripheral white blood cells in the treatment group were superior to those in the control group ( P0.05) .Conclusion Yougui Pills show better effect than its kidney-yang-warming separate formula for the treatment of kidney-yang deficiency syndrome, indicating that the method of generating yang by activating yin is superior to the method of simply nourishing kidney yang.
ABSTRACT
Objective: To observe the immunohistochemical expression of interferon-? (IFN-?), matrix metalloproteinase -9(MMP-9) in brain and spinal cord tissue, and explore the action mechanism of Zuogui Pills and Yougui Pills on rats with experimental autoimmune encephalomyelitis(EAE). Methods: Lewis rats were immunized with the myelin basic protein (MBP).120 Rats were grouped randomly into nomal group, EAE group, prednisone group, Zuogui Pills group and Yougui Pills group after post immunization (PI). Rats in normal group and EAE group were administered sodium, each 3ml/d, rats in prednisone-group were administered suspension of prednisone after developed clinical signs, each 5mg/kg, rats in Zuogui Pills group were administered suspension of Zuogui Pills, each 2g/kg, and rats in Yougui Pills group were administered suspension of Yougui Pills each 3g/kg. On days 15 and 27 PI, rats were killed and the immunohistochemical staining was performed on the sections of brain and spinal cord. Results: On days 15 and 27 PI, the expression of IFN-? and MMP-9 in central nervous system of EAE group were higher than those in nomal group (P