Pilose antler (Cervus elaphus Linnaeus) polysaccharide and polypeptide extract inhibits bone resorption in high turnover type osteoporosis by stimulating the MAKP and MMP-9 signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Pilose antler is a traditional Chinese medicine used to improve kidney function, strengthen tendons and bones, and prolong life, among other uses. It is widely employed in the treatment of osteoporosis. However, the molecular mechanisms underlying the treatment of high turnover osteoporosis are not fully understood. AIM OF THE STUDY: The present study aimed to investigate the molecular mechanism underlying pilose antler polysaccharide and polypeptide extracts in inhibiting bone resorption in high turnover osteoporosis, and compare the effects of the two components alone and in combination to explore whether they could produce synergistic enhancement effects. MATERIALS AND METHODS: The quantitative and qualitative characteristics of pilose antler polysaccharide and polypeptide extracts were detected by UV-visible spectrophotometry and high-performance liquid chromatography. A rat model of retinoic acid-induced osteoporosis was used to evaluate the inhibitory effect of the extracts on bone resorption. Enzyme-linked immunosorbent assay (ELISA) was used to detect the activity of factors related to high turnover type osteoporosis in rat serum. Western blotting was used to detect the expression of proteins related to the MAKP and MMP-9 signaling pathways in rat femurs. Fluorescence quantitative PCR was used to detect the transcription levels of genes related to the MAKP and MMP-9 signaling pathways in rat femur tissues. Hematoxylin and eosin staining were used to observe the osteoprotective effects of pilose antler polysaccharides and polypeptides. RESULTS: The yield of pilose antler polysaccharides was 8.3%, and was mainly composed of mannose, glucosamine hydrochloride, glucuronic acid, Galacturonic acid, Galactose hydrochloride, glucose, and galactose. The yield of the polypeptides was 26.2%, and eighty percent of the molecular weight of the antler polypeptides was 1.6 kDa-7kD, among which, the molecular weight of 7kD peptide accounted for 52% of the total. Both polysaccharides and peptides could reduce the activities of TRACP, OCN, ERK1, JNK, and MMP-9 in rat serum and reduce both the protein expression and gene transcription levels of ERK1, JNK, and MMP-9 in rat femur tissue with significant differences compared with the model group. Both extracts exerted significant protective effects on rat femur tissue. The effect of pilose antler polypeptides alone was better than that of polysaccharides either alone or in combination. CONCLUSIONS: Pilose antler polysaccharides, polypeptides, and their mixtures could inhibit the occurrence of bone resorption of high turnover osteoporosis by stimulating the MAKP and MMP-9 signaling pathways to reduce the expression of the ERK1, JNK, and MMP-9 genes and proteins, and could help alleviate bone loss caused by retinoic acid. Pilose antler polypeptides had a stronger effect on inhibiting bone resorption. The combination of the two components did not show synergistic enhancement effect, and the polysaccharide tended to moderate the inhibitory enhancement effect of the polypeptide.

[Qualitative and quantitative detection of Poria cocos by near infrared reflectance spectroscopy].

OBJECTIVE: The present study is concerning qualitative and quantitative detection of Poria cocos quality based on FT-near infrared (FT-NIR) spectroscopy combined with chemometrics. METHOD: The Poria cocos polysaccharides contents were determined by UV. Transmission mode was used in the collection of NIR spectral samples. The pretreatment method was first derivation and vector normalization. Then principal component analysis (PCA) was used to build classification model and partial least square (PLS) to build the calibration model. RESULT: The results showed that conventional criteria such as the R, root mean square error of calibration (RMSEC), and the root mean square error of prediction (RMSEP) are 0.944 0, 0.072 1 and 0.076 2, respectively. The misclassified sample is 0 using the qualitative model built by PCA. CONCLUSION: The prediction models based on NIR have a better performance with high precision, good stability and adaptability and can be used to predict the polysaccharose content of Poria cocos rapidly, which can provide a fast approach to discriminate the different parts of Poria cocos.

[Changes of fingerprints with crude and processed cassiae semen in xuezhiling tablets].

OBJECTIVE: To establish an effective and convenient method for HPLC fingerprints of Xuezhiling tablets and new Xuezhiling tablets, and to observe the changes of fingerprint with crude and processed Cassiae Semen in Xuezhiling tablets. METHODS: The HPLC with Agilent TC-C18 (4.6 mm x 250 mm, 5 microm) column was used for the gradient elution of acetonitrile-0.1% phosphoric acid solution, at the flow rate of 1.0 mL/min. Detection wavelength was set at 284 nm and the column temperature was 30 degrees C. RESULTS: Ten batches of Xuezhiling tablets and new Xuezhiling tablets were tested and gained HPLC fingerprint containing 20 common peaks, respectively. CONCLUSION: This method is stable and reliable. The number of common peaks of fingerprint had little change after the crude and processed Cassiae Semen in Xuezhiling tablets interchangeably, but the contents of some components had significant changes.

[Study on the tissue distribution of berberine from Rhizoma Coptidis and compatibility with Rhizoma Coptidis and Cortex Cinnamomi in rats].

OBJECTIVE: To study the effects of Rhizoma Coptidis extracts and Rhizoma Coptidis and Cortex Cinnamomi extracts on the distribution of terberine in rats' tissues. METHODS: The rats were intragastrically given the two different extracts. HPLC was used to determine the concentration of berberine in rats' tissues and the sample was extracted by liquid-liquid extraction method. RESULTS: At the same time, with the same gavage of the three extracts, the trend of the berberine in rats was: the concentration of Rhizoma Coptidis and Cortex Cinnamomi extracts > the concentration of Rhizoma Coptidis extracts. CONCLUSION: Berberine can be detected in intestinal, stomach, colon, pancreatic, heart, kidney, liver, spleen, lung, testicular, uterus, especially the gastrointestinal system with higher concentration, so berberine has a wide range of distribution in vivo.

[Studies on quality evaluation of Penis et Testis Cervi by Grey incidence degree method].

OBJECTIVE: To systematically evaluate the quality of Penis et Testis Cervi. METHODS: Grey model was set up by determining six main compositions contained in the samples. RESULTS: The quality evaluation on 9 samples by this model was as same as tradition and reality. CONCLUSION: Grey incidence degree method and the model can be used to evaluate the quality of Penis et Testis Cervi.

[Determination of oridonin and rosemarinic acid in Rabdosia rubescens by HPLC].

OBJECTIVE: To establish the method for determining the content of oridonin and rosemarinic acid in Rabdosia rubescens by HPLC. METHODS: Using ODS-C18 column (150 mm x 4.6 mm, 5 microm), methanol-0.3% phosphoric acid (40:60), oridonin detection wavelength at 238 nm and rosemarinic acid at 329 nm. RESULTS: The average recovery oridonin was 97. 1% and rosemarinic acid was 99.3%. Oridonin relative standard deviation (RSD) was 2.8% and rosemarinic acid was 2.9%. CONCLUSION: This method can be used to determine the content of oridonin and rosemarinic acid in Rabdosia rubescens.