Antioxidant and immunomodulatory activities of Oviductus ranae in mice

Oviductus ranae (OR) is a traditional Chinese medicine, which was first recorded in the Compendium of Materia Medica in the Ming Dynasty. OR contains high amounts of proteins and elicits therapeutic effects on neurasthenia, insomnia, and respiratory symptoms, which are related to oxidative stress and immunodeficiency. This study aimed to obtain the potential of OR for the development of functional food possessing antioxidant and immune-enhancement functions in the same dose. In antioxidant evaluation, OR can significantly decrease malondialdehyde and protein carbonyls (P < 0.05, P < 0.01) and significantly increase total superoxide dismutase and glutathione in a dose-dependent manner (P< 0.05, P < 0.01) against ethanol-induced oxidative stress in mice at 0.1, 1.0, and 3.0 g/kg BW. In immunomodulatory evaluation, OR could significantly enhance the phagocytosis of liver macrophages (P < 0.05, P < 0.01), delayed-type hypersensitivity response (P < 0.05, P < 0.01), hemolytic activity (P < 0.05), antibody-producing cells (P < 0.05), and natural killer cell activity (P < 0.05) in the same dose range described in antioxidant evaluation compared with those in the normal control. OR slightly influenced lymphocyte proliferation, peritoneal macrophage phagocytosis, and immune organ indices in mice. Thus, 3.0 g/kg BW OR showed potential for the development of functional food with antioxidant and immune-enhancement activities

Culture in vitro of rat ovarian granulosa cells and effect of Ranae Oviductus on its proliferation / 重庆医学

Objective To optimize the effective methods of isolation,purification,culture in vitro and identification of SD rat ovarian granulosa cells,to research the effects of Oviductus Ranae on the proliferation of rat ovarian granulosa cells by CCK-8,and to contrastively analyze the best optimal action concentration and time of serum contsining Oviductus Ranae on granulosa cells to lay the foundation for further in vitro experiment.Methods Nonage SD rats aged 25 d were selected and intraperitoneally injected by pregnant mare serum,then killed after 48 h.Ovarian granulosa cells were collected and cultured in the DMEM-F12 culture solution.The hematoxylin & eosin(HE) staining and immunofluorescence technique were used to identify the ovarian granulosa cells.Twen ty-five SD rats were randomly divided into the normal control group,positive medicine control group,and low,middle and high do ses Oviductus Ranae groups.Blood was collected and serum was separated after 7 d mediaction gavage.The volume percent of 10 ％,20％,40％,80％ serum in each group was added into the in vitro medium system of ovarian granulosa cells culture.Then the cell proliferation situation at 24,48,72 h in each group was measured by CCK-8.Results Oviductus Ranae significantly increased the proliferation ability of granulosa cells in a certain dose-dependent relation.With the increase of Oviductus Ranae concentration con centration,its.proliferation ability was gradually increased,after 48 h action,which in the Oviducthus Ranae-Comtaining serum group with the volume fraction of 20％ was most significant (P＜0.05).Conclusion Establishing in vitro cultural method of rat o varian granulosa cells is conductive to further research the action and mechanism of Oviductus Ranae on ovary.

Study on effect of Oviductus Ranae on the expression of p16, p21 and cyclinD1 in liver tissues of aged female rats / 中国药学杂志

OBJECTIVE: To investigate the effect of Oviductus Ranae (OR) on the expression of p16, p21 and cyclinD1 protein in liver tissue of D-galactose-induced aging female rats and to further explore the anti-aging mechanism of OR in female rats. METHODS: Fourty young female SD rats (SPF grade) were randomly divided into 5 groups including OR-H group, OR-M group, OR-L group, VE group, and D-gal group with 8 in each group. The 40 rats received subcutaneous injection of D-galactose on the nape for 42d to establish aging model. Another 8 rats were injected daily with normal saline (NS) at the same position as the normal control group. From the 15th day of the experiment, the rats were given oral OR (in OR group) or Vitamin E (in VE group) for 28 d. Then all the rats were sacrificed. P16 and p21 in liver tissue were detected by immuno-histochemistry (IHC), and cyclinD1 was detected by Western-blotting method. RESULTS: The result of IHC showed that p16 and p21 mainly multifocally located in endochylema of liver cells and their distribution was diffuse and focus. Positive cell integral of p16 and p21 in the liver tissue of D-gal female aged rats was significantly increased than in normal control group (P < 0.01). After taking OR, the integral of p16-positive cell in OR-M and OR-L groups was significantly decreased than in D-gal group (P < 0.01). The integral of p21-positive cell in OR-H, OR-M and OR-L groups was significantly decreased than D-gal group (P < 0.01). The result of Western-blotting showed that the expression of cyclinD1 protein in the liver tissue of D-gal female rats was significantly decreased than in normal control group (P < 0.01). The expression of cyclinD1 in all OR groups was significantly increased compared with D-gal group (P < 0.01). The expression of cyclinD1 in OR-H group was the highest in all OR groups. CONCLUSION: OR can decrease the over-expression of p16 and p21 protein in liver tissue. Meanwhile, OR can enhance the expression of cyclinD1 in the liver of female rats. OR can promote liver cell proliferation of aging female rats. The anti-aging effects of OR is possibly produced through regulation of p16, p21, and cyclinD1 signaling pathways to promote the expression of the proliferation regulatory protein, and the signaling pathways play an important role in delaying aging. Copyright 2012 by the Chinese Pharmaceutical Association.