Liuwei Dihuang Pills(六味地黄丸) Enhance the Effect of Western Medicine in Treating Type 2 Diabetes:A Meta-Analysis of Randomized Controlled Trials

Objective： To perform meta-analyses evaluating the efficacy of adding Liuwei Dihuang Pills （六味地黄丸 LDP） to Western medicine in improving treatment outcomes for type 2 diabetes. Methods： Medline, PubMed, Cochrane Library, and Chinese databases, including the Chinese National Knowledge Infrastructure were searched to identify eligible studies; i.e., if the study involved a randomized clinical trial in which the experimental group combined LDP with Western drugs and the control group used the corresponding Western drugs alone to treat type 2 diabetes. Outcomes were measured in terms of fasting blood glucose （FBG）, postprandial blood glucose （2hPG） and HbAlc level. Efficacy was also measured by using control and response rates. The combined odds ratio （OR）, mean difference （MD）, and 95% confidence intervals （95% CI） were calculated. Results： Studies included in the analysis were less adequate than expected in terms of methodological qualify. A total of 1,609 patients from 18 studies were included. We found that adding LDP can lower patients＇ FBG （MD=0.54 mmol/L, 95% CI [0.15, 0.93], P=0.007）, 2hPG （MD=1.05 mmol/L, 95% CI [0.29, 1.81], P〈0.01） and HbAlc （MD=0.23, 95% CI [0.02, 0.45], P=0.008）. There were also improvements in treatment response rates （OR=3.41, 95% CI [2.38, 4.90], P〈0.01） and control rates （OR=2.47, 95% CI [1.91, 3.20], P〈0.01）. Conclusion： Adding LDP to Western medicine might improve treatment outcomes of diabetes, including FBG, 2hPG, response rates and control rates.

Inhibition of N-methyl-N-nitrosourea-induced gastric tumorigenesis by Liuwei Dihuang Pill in db/db mice

AIM To investigate the inhibitory effect of Liuwei Dihuang Pill(LDP) on gastric tumorigenesis induced by N-methyl-N-nitrosourea(MNU) in diabetic mice.METHODS Four-week-old mice were divided into four groups: A, 12 db/m mice treated with MNU and saline, as the non-diabetic control; B, 12 db/db mice treated with MNU and saline, as the diabetic control; C, 12 db/db mice treated with MNU and metformin, as the positive control; and D, 12 db/db mice treated with MNU and LDP. MNU was administrated for 20 wk to induce gastric carcinogenesis. LDP was administrated for 10 wk for improvement of insulin resistance. Body weight and food intake were measured every week. Blood samples were collected for assays of fasting blood glucose, insulin, insulin-like growth factor(IGF)-1, adiponectin and leptin. Stomach tissues were collected for histopathological analysis, immunohistochemical staining of Ki67, quantitative reverse transcriptionpolymerase chain reaction and western blotting. RESULTS The incidence of MNU-induced gastric dysplasia was significantly elevated in diabetic(db/db) mice relative to the control(db/m) mice. The incidence of gastricdysplasia was significantly reduced by LDP with suppression of cell proliferation, as demonstrated by a decrease in Ki67 staining. Hyperglycemia, hyperinsulinemia and serum IGF-1 were inhibited by LDP. Expression of IGF-1 and insulin receptor m RNAs was decreased, phosphorylation of IGF-1 receptor and AKT protein was reduced in the stomach tissues by LDP. In addition, adiponectin was increased and leptin was decreased in the serum by LDP. CONCLUSION LDP decreased risk of gastric dysplasia in type 2 diabetic mice by down-regulation of IGF and insulin activity and correction of adipokines disorders.

Changes of Leucocytic Estrogen Receptor Levelsin Patients with Climacteric Syndrome andthe Therapeutic Effects of LiuweiDihuang Pills (六味地黄丸)

he levels of estrogen receptor (ER) in human peripheral leucocytes in 22 women withclimacteric syndrome were measured by radioligand method. The results were compared with those of 12healthy women of child-bearing age. It was found that the contents of leucocytic ER in patients with cli-macteric syndrome were significantly lower than healthy women of child-bearing age (372±149 vs 1143±255 Rs binding sites/cell). The authors used a Chinese prescription---Liuwei Dihuang Pills (LDP) totreat the patients for 2 months. The levels of leucocytic ER were increased affer treatment. The data indi-cate a decrease of ER Ievels in leucocytes may be involved in the pathogenesis of climacteric syndrome.LDP not only increase plasma estradiol levels , but also increase the leucocytic ER levels. This may be thebasis of the therapeutic effect of LDP on climacteric syndrome.

Liuwei Dihuang Pill(六味地黄丸)Treats Postmenopausal Osteoporosis with Shen(Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression

Objectives： To investigate the mechanism of Liuwei Dihuang Pill （六味地黄丸, LDP） in treating postmenopausal osteoporosis （PMOP） with Shen （Kidney） yin deficiency. Methods： In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group （Group A）, PMOP Shen-yang deficiency group （Group B）, PMOP without Shen deficiency group （Group C）, and control group （Group N）. Real-time polymerase chain reaction （RT-PCR） and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 （CLCF1）, ankyrin repeat and SOCS box containing 1 （ASB1）, and proldneticin 2 （PROK2） genes and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. Results： The mRNA （P〈0.05） and protein （P〈0.01） expression levels of the CLCF1 gone in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gone were obviously up-regulated （P〈0.01）. After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated （both P〈0.01）, and the average bone density of the top femur had significantly increased （P〈0.05）. In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3. Conclusions： The CLCF1 gone is an important gone associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gone expression and activation of the JAK/STAT signaling pathway.

Liuwei Dihuang pill suppresses metastasis by regulating the wnt pathway and disrupting β-catenin/T cell factor interactions in a murine model of triple-negative breast cancer

OBJECTIVE:To investigate if the Liuwei Dihuang pill(LWDHP)can inhibit metastasis to the liver and lungs in mice bearing triple-negative breast cancer(TNBC),and the molecular mechanism underpinning this action.METHODS:Ninety-nine TNBC bearing-mice were distributed randomly to five groups:control(Con),paclitaxel(PTX),low-dose LWDHP(LLP,2.3 g·kg^-1·d^-1),middle-dose LWDHP(MLP,4.6 g·kg^-1·d^-1)and high-dose LWDHP(HLP,9.2 g·kg^-1·d^-1).The LWDHP were administered(p.o.)to the agonal stage.The morphology of BC cells was observed by hematoxylin&eosin staining.Expression of axin-2,β-catenin,T cell factor(TCF),cyclin-D1 and vascular endothelial growth factor(VEGF)was detected by western blotting or immunofluorescence.β-catenin/TCF-1 interaction was measured using a co-immunoprecipitation assay.RESULTS:After LWDHP treatment,metastasis of BC cells to the lungs and liver was inhibited,expression of axin-2 was increased,expression of TCF-1,β-catenin,cyclin-D1 and VEGF was decreased,andβ-catenin/TCF-1 interaction was disrupted.CONCLUSION:The LWDHP could inhibit metastasis of BC cells to the liver and lungs.The molecular mechanism underlying this action may be regulation of protein expression andβ-catenin/TCF-1 interactions in the Wnt pathway.

Contribution of Borneolum syntheticum to the Intervention Effect of Liuwei Dihuang Pill (六味地黄丸) on Experimental Retinal Degeneration

Objective： To observe the contribution of Borneolum syntheticum to the intervention effect of Liuwei Dihuang Pill （六味地黄丸, LDP） on experimental retinal degeneration, and initially investigate the mechanism of Borneolum syntheticum as meridian-lead-in drug. Methods： A total of 180 sodium iodate- induced retinital degeneration rats were randomly divided into three groups, including distilled water group, LDP group, and LDP＋Borneolum syntheticum （LDP＋BS） group. Twenty normal rats were fed regularly without any treatment as normal control. On day 7 and 14 after treatment, histopathological study and transferase-mediated deoxyuridine triphosphate-biotin nick end labeling （TUNEL） test were performed to evaluate the retinopathy. Claudin-5 expression at blood-retina barrier （BRB） was detected by Western blot at different time points from 0.5 to 8 h after gavage. Results： On day 7 and 14 after treatment, the retinal lesion grades were significantly different among the three groups （P〈0.05）. The grade in the LDP＋BS group was significantly less than the LDP and distilled water groups （both P〈0.05）, no significant difference was observed between the LDP and distilled water groups （P〉0.05）. The apoptosis rates in the LDP＋BS group was significantly less than the distilled water and LDP groups （both P〈0.05）, while there was no significant difference between LDP and distilled water groups （P〉0.05）. Expression of claudin-5 in LDP＋BS group was significantly less than the other two groups at 0.5, 1 and 2 h after gavage （P〈0.05）. There was no apparent difference among the three groups at 4 and 8 h after gavage （P〉0.05）. Conclusion： Bomeolum syntheticum could strengthen the effect of LDP on experimental retinal degeneration, indicated that Bomeolum syntheticum might play the role of meridian-lead-in drug in the formula. The mechanism may be due to Bomeolum syntheticum could promote the physiologically openness of blood- retina barrier through transiently affecting the expression of claudin-5.