This study investigated the inhibitory effect of grape seed proanthocyanidin extract (GSPE) on selenite-induced cataract formation in rats and the possible mechanism. Eighty 8-day-old Sprague-Dawley rats were divided randomly into 5 groups: control group, model group, three GSPE groups (low dose, medium dose and high dose). Control group received subcutaneous injection of physiological saline. Model group was given subcutaneous injection of sodium selenite (20 μmol/kg body weight) on the postpartum day 10, and once every other day for consecutive three times thereafter. GSPE treated groups were respectively administered GSPE at doses of 50, 100, and 200 mg/kg body weight intragastrically 2 days prior to the selenite injection (that was, on the postpartum day 8), and once daily for fourteen consecutive days thereafter. The opacity of lenses was observed, graded and photographed under the slit lamp microscopy and the maximal diameter of the nuclear cataract plaques was measured. The lenses were analyzed for superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX), malondialdehyde (MDA), calcium (Ca(2+)), nitric oxide (NO) and anti-hydroxyl radical ability (anti-OH(-)). The histomorphology of lenses was observed with HE staining under a light microscope. The levels of calpainII, and iNOS protein and mRNA expression in lenses were detected by using immunohistochemistry and real-time quantitative RT-PCR. The results showed subcutaneous injection of sodium selenite led to severe nuclear cataract in model group, and the achievement ratio of model group was 100%. As compared with model group, the degree of lenses opacity and the maximal diameter of nuclear cataract plaques were significantly reduced in GSPE-treated groups. Moreover, we observed selenite treatment caused a significant decrease in the activities of antioxidative enzymes (SOD, CAT, GSH-PX) and anti-OH(-) ability, accompanied by a significant increase in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. Administration of GSPE could dose-dependently preserve the activities of these antioxidative enzymes and anti-OH(-) ability, accompanied by a significant reduction in the levels of MDA, NO, Ca(2+) as well as iNOS, and calpainII protein and mRNA expression. These results suggested that GSPE markedly prevented selenite-induced cataract formation probably by suppressing the generation of lipid peroxidation and free radicals as well as the activation of iNOS, and calpainII in the lenses.
Animals ; Cataract ; chemically induced ; drug therapy ; Grape Seed Extract ; pharmacology ; Plant Extracts ; pharmacology ; Proanthocyanidins ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Selenious Acid ; adverse effects

OBJECTIVETo determine the difference on purgative biopotency of different processed products of rhubarb and compare rhubarb before and after preparation.

METHODThe prime biopotency of rhubarb reference substance was determined by comparing with the control substance of sennoside B using rat purgative model. Comparing with calibrated rhubarb reference substance, the bioactivity different processed products of rhubarb was determined by the 2,2',2" method.

RESULTThe purgative biopotency of crude rhubarb was 825.22 U x g(-1). The purgative biopotency of alcohol-processed rhubarb was 699.05 U x g(-1), The purgative biopotency of steamed rhubarb were 459.76 U x g(-1). Carbonized rhubarb cannot be determinate.

CONCLUSIONCrude rhubarb showed a significantly declineing purgative biopotency after being processed, alcohol-processed rhubarb showed less than crude rhubarb and followed by steamed rhubarb. Carbonized rhubarb lost almost all purgative bioactivity.

Animals ; Cathartics ; administration & dosage ; pharmacology ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Male ; Mice ; Mice, Inbred ICR ; Plant Extracts ; administration & dosage ; pharmacology ; Rheum ; chemistry ; Senna Extract ; pharmacology

BACKGROUNDDiabetic retinopathy (DR) is a leading cause of visual impairment and blindness among the people of occupational age. To prevent the progress of retina injury, effective therapies directed toward the key molecular target are required. Grape seed proanthocyanidin extracts (GSPE) have been reported to be effective in treating diabetic complications, while little is discussed about the functional protein changes.

METHODSWe used streptozotocin (STZ) to induce diabetes in rats. GSPE (250 mg/kg body weight per day) were administrated to diabetic rats for 24 weeks. Serum glucose, glycated hemoglobin and advanced glycation end products (AGEs) were determined. Consequently, 2-D difference gel electrophoresis and mass spectrometry were used to investigate retina protein profiles among control, STZ-induced diabetic rats, and GSPE treated diabetic rats.

RESULTSGSPE significantly reduced the AGEs of diabetic rats (P < 0.05). Moreover, GSPE significantly suppressed the vascular lesions of central regions, decreased capillary enlargements and neovascularization, similar to those of the control rats under light microscope. Eighteen proteins were found either up-regulated or down-regulated in the retina of STZ-induced diabetic rats. And seven proteins in the retina of diabetic rats were found to be back-regulated to normal levels after GSPE therapy. These back-regulated proteins are involved in many important biological processes such as heat shock, ubiquitin-proteasome system, cell proliferation, cell growth and glucose metabolism.

CONCLUSIONSThese findings might promote a better understanding for the mechanism of DR, and provide novel targets for evaluating the effects of GSPE therapy.

Animals ; Blood Glucose ; drug effects ; metabolism ; Body Weight ; drug effects ; Diabetes Mellitus, Experimental ; complications ; metabolism ; pathology ; Diabetic Retinopathy ; drug therapy ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Glycated Hemoglobin A ; metabolism ; Glycation End Products, Advanced ; metabolism ; Grape Seed Extract ; Male ; Plant Extracts ; pharmacology ; Proanthocyanidins ; pharmacology ; Proteomics ; methods ; Rats ; Rats, Wistar

Background: Pectin is a pharmaceutically relevant excipient that can be upcycled from selected Philippine fruit peel wastes. Method optimization of pectin extraction leads to maximizing yields from limited resources, while also reducing environmental wastes, and providing local alternative sources. Objectives: This study aimed to optimize the method of extracting pectin from selected Philippine fruit peel wastes using the Box-Behnken design, by varying the acid extraction solvent, treatment time, and working temperature. Methodology: The three-level (-1, 0, 1) Box-Behnken design (15 set-ups) was used to optimize the pectin extraction in each of the fruit peel samples (C. maxima; A. heterophyllus; ripe and unripe M. indica; D. zibethinus; and H. undatus). The three experimental factors were the type of 3N acid used as extracting solvent (HNO₃, H₂SO₄, and HCl); duration of treatment in minutes (60, 90, and 120); and temperature of treatment in C 60, 75, and 90). The %yield was computed in each set-up, and the projected yields were generated using multiple linear regression. The pectin samples obtained from the optimized conditions were subjected to the physicochemical characterization, with apple pectin as the standard. Degree of esterification (DE), equivalent weight (EW), methoxy content (MC), alkalinity of ash (AA), and anhydrouronic acid content (AUA) were performed. Results: Maximum yields were extracted from C. maxima (28.96%), A. heterophyllus (20.12%), ripe M. indica (26.23%), and unripe M. indica (25.89%), using 3N H₂SO₄, for a treatment duration of 60 minutes, at a working temperature of 90 C, and H. undatus (25.03%) at 60 C, for a treatment duration of 120 minutes. Conclusion Optimum conditions were identified to extract pectin in each of the fruit peel samples. The 3N H₂SO₄ produced the highest pectin yields in all of the set-ups, while the treatment time and working temperature vary per fruit peel sample. Pectin extract from C. maxima, A. heterophyllus, and M. indica was comparable to the standard.
Pectins

The aim of this study was to investigate the protective effects of compound tianpupian (TPP) and its compositions against oxidative damage in mouse erythrocytes. The protective effect of TPP and its compositions against the red cell hemolysis induced by (2)O(2) or auto-oxidation were observed by scanning electron microscopy and spectrophotomety. The result indicated that compound TPP and all of its four components including extract of Rhodiola sachalinensis, Grape Seed Extract proanthocyanidins, Acanthopanax senticosus extract, and tea polyphenols had significant inhibitory activities for the oxidative damage of mouse erythrocytes, out of which the Grape seed extract proanthocyanidins showed the maximal protective effect. It is concluded that compound TPP can protect erythrocytes against oxidative stress and can be used as a valuable Chinese traditional medicine.
Animals ; Antioxidants ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Erythrocytes ; drug effects ; metabolism ; Grape Seed Extract ; pharmacology ; Hemolysis ; drug effects ; Mice ; Mice, Inbred ICR ; Oxidation-Reduction ; Oxidative Stress ; drug effects ; Proanthocyanidins ; pharmacology ; Rhodiola

The aim of this study was to explore the protective effect of compound tianpupian (TPP) against (2)O(2)-induced the apoptosis of murine splenic lymphocytes and its mechanism. The cell apoptosis rate was detected by MTT method; the cell apoptosis and mitochondrial membrance potential were detected by flow cytometry (FCM) with Annexi-V/PI double staining and JC-1 staining method, respectively; and caspase 3 relative activity was determined by colorimetry. The results indicated that after treating with (2)O(2), the absorbance value of cultured lymphocytes and the red/green ratio of JC-1 were reduced, and the apoptotic rate and caspase 3 activity were increased, coculture of (2)O(2)-treated cells with compound TPP increased the cell absorbance ratio and red/green rate of JC-1, while reduced the apoptosis rate and caspase 3 activity. It is concluded that compound TPP alleviates intracellular oxidative damages and dose-dependently inhibited apoptosis of murine splenic lymphocytes through reducing mitochondrial membrane potential and inhibiting caspase 3 activity. This suggests that compound TPP is a potential anti-apoptotic agent.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Grape Seed Extract ; pharmacology ; Hydrogen Peroxide ; adverse effects ; Lymphocyte Count ; Lymphocytes ; cytology ; drug effects ; Mice ; Mice, Inbred ICR ; Proanthocyanidins ; pharmacology ; Rhodiola

Paeoniae Radix Alba is the dried root of Paeonia lactiflora, which was first recorded in the Shennong's Classic of Materia Medica and listed as the top grade. It is a common blood-tonifying herb, and its chemical components are mainly monoterpenes and their glycosides, triterpenes, flavonoids and so on. Modern research has demonstrated that Paeoniae Radix Alba has the activities of anti-inflammation, pain easing, liver protection, and anti-oxidation, and thus it is widely used in clinical practice and has broad development prospects. In this paper, the research progress on the chemical composition, pharmacological effects, and quality control of Paeoniae Radix Alba were summarized. On this basis, the Q-markers of Paeoniae Radix Alba were predicted from the aspects of mass transfer and traceability, chemical composition specificity, and availability and measurability of chemical components, which will provide a scientific basis for the quality evaluation of Paeoniae Radix Alba.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Monoterpenes ; Paeonia ; Plant Extracts

As a most important Chinese materia medica, Aconiti Lateralis Radix Praeparata(Fuzi) had been widely used in China for thousands of years. This herbalogical study was systematically performed based on variation characteristics of the naming, habitat, harvesting, processing and properties. The sharp toxicity of Fuzi had been well known since the spring and autumn period in the history, which was much earlier than that its medical properties was understood and applied. Sichuan province was regarded as the geo-authentic region of Fuzi all along, where the best quality goods could be provided for clinic use. The study showed the harvesting time of Fuzi was changing in different periods, and the possible effects were of climate change and artificial planting. The perishable characteris-tics of Fuzi severely limited its storage period; therefore, different kinds of storage methods were effectively used since Tang Dynasty. For thousands of years, Fuzi had been processed with various accessories to reduce toxicity, while simultaneously the study on processing mechanism was on going all the time. Fuzi was widely used in clinical practice to cure Yang depletion syndrome, which was based on its function of enhancing Yang and removing cold. Along with the further study on quality evaluation standard, Fuzi will probably get a much wider range of applications.
Aconitum ; China ; Drugs, Chinese Herbal/toxicity* ; Plant Extracts

In the pharmacopoeia, many process parameters for the purification process of Scutellariae Radix are unclear. In this study, deterministic screening design combined with design space method was used to optimize the purification process of Scutellariae Radix extract. Nine method parameters such as mass fraction of solution(X_1), first acid precipitation pH(X_2) and first holding time(X_3) in the purification process were firstly studied by definitive screening design. The yield of baicalin was defined as the evaluation index. A stepwise regression method was used then to build quantitative models between evaluation index and method parameters and the three most critical impact parameters were determined. Probability-based design space was calculated and successfully verified with the experimental error simulation method. Finally, the second standing temperature, the first standing temperature and the pH value of the second acid precipitation were determined as the three most critical method parameters. The recommended operating space was as follows: the second standing temperature 5-7 ℃, the first standing temperature 13-15 ℃, and the pH of the second acid precipitation 1.5-1.7. Within this operating space, the baicalin yield in the purification process was over 80%, and the probability of reaching the standard was over 0.96. In this study, we optimized the effect of various parameters for the purification process of the Scutellariae Radix extract in the pharmacopoeia on the yield of baicalin and provided a reference for industrial production of the exact of Scutellariae Radix.
Drugs, Chinese Herbal ; Flavonoids ; Plant Extracts ; Scutellaria baicalensis

With the rice-steamed Rehmanniae Radix unearthed from the tomb of Haihunhou in the Western Han Dynasty as the re-ference, the present study evaluated the quality of Rehmanniae Radix and investigated the processing technology of rice-steamed Rehmanniae Radix to lay the foundation for the research on rice-steamed Rehmanniae Radix products. With catalpol and rehmannioside D as the investigation indexes, the quality and grade of Rehmanniae Radix from different producing areas were evaluated with the methods in 2020 edition of Chinese Pharmacopoeia. UPLC method was established for the determination of catalpol and rehmannioside D in the rice-steamed Rehmanniae Radix. The effects of steaming time, the amount of supplementary rice, and steaming times in the rice-steamed processing on the quality of products were investigated by L_9(3~4) orthogonal test and multi-index comprehensive balance scoring method combined with the content of catalpol and rehmannioside D and appearance characteristics. At last, the stability of the processing technology was tested. The results showed that the optimal processing technology for rice-steamed Rehmanniae Radix was as follows: Rehmanniae Radix and rice(200 g∶4 g) were steamed twice at atmospheric pressure, four hours each time. The mass fractions of catalpol and rehmannioside D were 0.184% and 0.335%, respectively, and the character score was 6.5. The processing conditions are reaso-nable, stable, and feasible. It can provide a basis for the restoration of the ancient rice-steamed processing technology and references for the development of rice-steamed Rehmanniae Radix products in the future.
Drugs, Chinese Herbal/pharmacology* ; Oryza ; Plant Extracts ; Rehmannia ; Technology