[Mechanism of Danzhi Jiangtang Capsules on improving liver injury in hyperlipidemia rats based on MAPK pathway].

This study aimed to investigate the protective effect and preliminary mechanism of Danzhi Jiangtang Capsules( DJC) on liver of hyperlipidemic rats. The hyperlipidemia models were successfully made by high-fat diet for 12 weeks in male SD rats,and then divided into model control group and DJC treatment groups( 500 and 1 000 mg·kg~(-1)·d-1) via gavage administration for additional 8 weeks.The levels of serum lipid and liver metabolism indices were detected; HE and oil red O staining were used to observe the pathological changes of liver. Expression levels of extracellular regulated protein kinase 1/2( ERK1/2),c-Jun N-terminal kinase( JNK),and p38 mitogen-activated protein kinase( p38 MAPK) were detected by real-time polymerase chain reaction( RT-PCR). Expression of MCP-1,phosphorylated ERK( p-ERK),phosphorylated JNK( p-JNK),and phosphorylated p38 MAPK( p-p38) were analyzed by immunohistochemistry and Western blot. The results showed that DJC decreased body weight and serum levels of total cholesterol( TC),triglyceride( TG),alanine aminotransferase( ALT),aspartate aminotransferase( AST),increased serum high-density lipoprotein cholesterol( HDL-C) level,ameliorate injury and lipid deposition in the liver induced by the high-fat diet,decreased mRNA expression of ERK1/2,JNK and p-38 MAPK as well as protein expression of p-ERK,p-JNK,p-p38,and MCP-1,somewhat showing a dose-dependent effect. Therefore,DJC has an obvious protective effect on liver of hyperlipidemic rats with certain dose-dependent effect,and the mechanism may be related with inhibiting MAPK pathways and inflammation.

[Effect of Active Ingredients of Qingxin Kaiqiao Recipe on Spatial Learning and Memory Capacities, and Expressions of Bax, Bcl-2, Caspase-3, and PAPP in Hippocampus of AD Model Rats].

Objective To observe the effects of active ingredients of Qingxin Kaiqiao Recipe (QKR) , such as saponins, volatile oils, effective compositions of polysaccharides, on expressions of Bcl-2 associated X protein (Bax) , B-cell lymphoma-2 (Bcl-2) , cysteinyl aspartate specific proteinase-3 (Caspase-3) , and ß-amyloid precursor protein (pAPP) in hippocampus of Ap1_40-induced Alzheimer's disease (AD) model rats. Methods Totally 112 male Sprague-Dawley (SD) rats were randomly divided into 7 groups, i.e., the normal control group, the sham-operation group, the model group, the Aricept group, the saponin group, the volatile oil group, the polysaccharide group, 16 in each group. The AD rat model was established by injecting Aß1₋40 from bilateral hippocampus. Equal volume of double distilled water was administered to rats by gastrogavage in the normal control group, the sham-operation group, the model group from the 2nd day after modeling, once per day for 2 successive weeks (at 10:00 am). Aricept (Donepezil Hydrochloride Tablet, 1. 67 mg/kg per day) , saponin (9 mL/kg per day) , benzene (3. 33 mL/kg per day) , and polysaccharides (8. 33 mL/kg per day) was administered to rats by gastro- gavage to the Aricept group, the saponin group, the volatile oil group, the polysaccharides group, re- spectively, once per day for 2 successive weeks (at 10:00 am). By the end of gastrogavage spatial learning and memory capacities were detected using Morris water maze (MWZ). Apoptosis in hippocam- pal CAI region was detected using TUNEL staining. Expressions of Bax, Bcl-2, Caspase-3, and PAPP were measured via Real-time fluorescent quantitative PCR, Western blot, and immunohistochemistry, respectively. Results There was no statistical difference in pre-modeling escape latency and times of crossing platforms among groups at the same time point (P >0. 05). Besides, escape latency was gradu- ally shortened as time went by. Compared with the model group, escape latency was shortened, and times of crossing platforms was significantly increased in the Aricept group and the saponin group (P < 0. 05, P <0. 01). Compared with the model group, the amount of apoptotic cells in hippocampal CA1 re- gion was obviously reduced (P <0. 05, P <0. 01) , expressions levels of Bax, Caspase-3, and pAPP were down-regulated, Bcl-2/Bax ratio was obviously elevated in the saponin group, the volatile oil group, the polysaccharide group (P <0. 05, P <0. 01). Conclusion Three active ingredients (spaonins, benzene, and polysaccharides) of QKR could improve spatial memory and learning capacities to different degrees, which might be possibly achieved by decreasing expressions of Bax, Caspase-3, PAPP in hippocampal CA1 region, elevating Bcl-2 expression, and inhibiting apoptosis in hippocampus.

Development and Characterization of Polymorphic Microsatellite Markers for Sedum sarmentosum (Crassulaceae) and Their Cross-Species Transferability.

Sedum sarmentosum is an important Chinese medicinal herb that exhibits anti-inflammatory, anti-angiogenic and anti-nociceptive properties. However, little is known about its genetic background. The first set of 14 microsatellite markers were isolated and characterized for S. sarmentosum using an SSR-enriched library. Fourteen polymorphic microsatellite markers were acquired with satisfactory amplifications and a polymorphic pattern in 48 S. sarmentosum individuals. The number of alleles ranged from 3 to 15. The observed and expected heterozygosities varied from 0.0833 to 0.8750 and 0.2168 to 0.9063, respectively. Two loci showed significant departure from the Hardy-Weinberg equilibrium. Cross-species amplification was carried out in other Sedum species. High rates of cross-species amplification were observed. The transferability value ranged from 85.7% in S. lineare to 64.3% in S. ellacombianum. These markers will be valuable for studying the genetic variation, population structure and germplasm characterization of S. sarmentosum and related Sedum species.

[Study on the Mechanism of Three Kinds Extracts of Qingxin Kaiqiao Recipe in Improving Learning and Memory Capabilities of AD Rats].

OBJECTIVE: To explore the mechanism of three kinds extracts (saponins, volatile components, polysaccharide components) of Qingxin Kaiqiao Recipe (QKR) in improving learning and memory capabilities of Alzheimer's disease (AD) rats. METHODS: A controlled comparison method was used. Totally 56 male SD rats were randomly divided into seven groups, i.e., the normal control group, the sham-operation group, the model group, the Aricept group, the saponin group, the benzene group, and the polysaccharide group, 8 in each group. AD rat model was established by bilateral hippocampus injection of Aß1-40 (2 µL, 2.5 µg/µL). The next day after modeling rats in the saponin group, the benzene group, and the polysaccharide group, the saponin group, the Aricept group were intragastrically administered with saponin (at the daily dose of 9 mL/kg, 2.1 g/mL) , benzene (at the daily dose of 3.33 mL/kg, 5.7 g/mL) , polysaccharide (at the daily dose of 8.33 mL/kg, 2.28 g/mL), Aricept (at the daily dose of 1.67 mg/kg), respectively, once a day for 2 consecutive weeks from 10 am every day. Equal volume of normal saline was intragastrically administered to rats in the normal control group and the model group. Learning and memory capabilities were detected using water maze 2 weeks later. Expression levels of synaptotagmin-1 (Syt-1), interleukin-1ß (IL-1ß), glia fibrillary acidic protein (GFAP), and ß-amyloid precursor protein (ßAPP) in the cortex and hippocampus of AD rats were detected using immunohistochemistry. RESULTS: Learning and memory capabilities could be improved by three kinds extracts of QKR. There was no statistical difference in the escape latency between the polysaccharide group and the model group (P >0. 05). The escape lacency was shortened in the rest treatment groups (P < 0.05). The escape latency was obviously prolonged in three kinds extracts of QKR groups, when compared with the Aricept group (P < 0.05, P < 0.01). Compared with the model group, times for crossing platforms were significantly increased in the saponin group and the Aricept group (P < 0.05). Compared with the Aricept group, average times for crossing platforms were significantly lessened in three kinds extracts of QKR groups (P < 0.01). Compared with the sham-operation group, expression levels of Syt-1, IL-1ß, GFAP, and ßAPP in the cortex and hippocampus were increased in the model group (P < 0.01). Compared with the model group, the expression of cortical Syt-1 increased in the saponin group and the benzene group; the expression of cortical IL-1ß increased in the benzene group and the polysaccharide group; the expression of hippocampal GFAP increased in the three kinds extracts of QKR groups; expression levels of Syt-1, IL-1ß, GFAP, and ß-APP in the cortex and hippocampus decreased in the rest treatment groups (all P < 0.05, P < 0.01). Compared with the Aricept group, expression levels of Syt-1, IL-1ß, GFAP, and ßAPP in the cortex and hippocampus were significantly increased in three kinds extracts of QKR groups (P < 0.05, P < 0.01). CONCLUSION: Three kinds extracts of QKR might play roles in anti-AD possibly by decreasing expression levels of Syt-1, IL-1ß, GFAP, and ßAPP in the cortex and hippocampus.

Distinct effects of naringenin and hesperetin on nitric oxide production from endothelial cells.

Diets rich in citrus and citrus-based products have been negatively correlated with the risk of cardiovascular disease, but so far no studies have been conducted to determine whether naringenin and hesperetin, two major flavanones in citrus plants, influence endothelium nitric oxide (NO) production. The aim of this study is to clarify estrogenic activities of naringenin and hesperetin and to examine whether they affect endothelial NO production via estrogen receptor (ER) activation. Both naringenin and hesperetin were observed to promote growth of MCF-7 cells under greatly reduced estrogen conditions and to suppress estrogen-induced response. Naringenin activated both ERalpha and ERbeta, whereas hesperetin exhibited stronger potential to activate ERalpha rather than ERbeta. Hesperetin, but not naringenin, increased NO releases from human umbilical vein endothelial cells in a dose-dependent manner. Hesperetin-induce responses were suppressed by ICI 182 780 and actinomycin D. Real-time reverse transcription polymerase chain reaction (RT-PCR) and western-blotting analysis revealed that hesperetin up-regulated endothelium nitric oxide synthase (eNOS) expression. These results suggested that hesperetin exerts an antiatherogenic effect, in part, via ER-mediated eNOS expression and subsequent increase of endothelial NO production. Distinct effects of naringenin and hesperetin on NO production also imply that ERalpha might play the major role in estrogen-induced eNOS expression. However, the inefficacy of naringenin on NO production remains to be elaborately studied. Our findings add more proof to the molecular explanations for the health benefits of citrus used to prevent cardiovascular disease, especially for postmenopausal women.

[Effects of yishen huoxue decoction on proliferating cell nuclear antigen expression and extracellular matrix in 5/6 nephrectomized rats].

OBJECTIVE: To investigate the effects and mechanisms of Yishen Huoxue decoction (YHD) on chronic renal failure (CRF) rats induced by 5/6 nephrectomy. METHODS: The glomerulosclerosis model was established by 5/6 nephrectomy in rats. Experimental animals were allocated into the normal group, the model group, the YHD group and the benazepril group. Urine protein of 24 h (UP) at the 6th and 12th weekend after operation, blood urea nitrogen (BUN) and creatinine (SCr), albumin (Alb) and haemoglobin (HB) at the 12th weekend were measured, renal pathology changes were examined with light microscope, the expressions of proliferating cell nuclear antigen (PCNA) and fibronectin (Fn) were examined by immunohistochemistry and mRNA expressions of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) by RT-PCR at the 12th weekend. RESULTS: Compared with those in the normal group, the levels of UP, BUN and SCr, the area of glomerular mesangial matrix, the FN deposition, PCNA expression in glomeruli and tubular interstitium and mRNA expressions of CTGF and PAI-1 were all significantly higher in the model group (P < 0.05). All the above-mentioned indexes were lower in the YHD group than those in the model group (P < 0.05). PCNA positively expressed cells in glomeruli of the normal, model group, YHD group and benazepril group was 7.00 +/- 2.24,34.78 +/- 6.96,15.75 +/- 2.61 and 15.50 +/- 2.57 respectively, positively correlated to the expression of CTGF, PAI-1, FN and SCr level. CONCLUSION: YHD could delay the progression of CRF in 5/6 nephrectomized rats, and the mechanisms were mainly related to the inhibition on renal cell proliferation and it induced over-expression of cytokines, and accumulation of extracellular matrix.

[Spectroscopic analysis of the interaction of ethanol and acid phosphatase from wheat germ].

Conformational and activity changes of acid phosphatase from wheat germ in ethanol solutions of different concentrations were measured by fluorescence spectra and differential UV-absorption spectra. The effect of ethanol on kinetics of acid phosphatase was determined by using the double reciprocal plot. The results indicate the ethanol has a significant effect on the activity and conformation of acid phosphatase. The activity of acid phosphatase decreased linearly with increasing the concentration of ethanol. Differential UV-absorption spectra of the enzyme denatured in ethanol solutions showed two positive peaks at 213 and 234 nm, respectively. The peaks on the differential UV-absorption spectra suggested that the conformation of enzyme molecule changed from orderly structure to out-of-order crispation. The fluorescence emission peak intensity of the enzyme gradually strengthened with increasing ethanol concentration, which is in concordance with the conformational change of the microenvironments of tyrosine and tryptophan residues. The results indicate that the expression of the enzyme activity correlates with the stability and integrity of the enzyme conformation to a great degree. Ethanol is uncompetitive inhibitor of acid phosphatase.