Zengsheng Han

Antidiabetic and antioxidant effects of oleanolic acid from Ligustrum lucidum Ait in alloxan-induced diabetic rats.

The present study evaluated the antidiabetic and antioxidant effects of oleanolic acid (OA) from Ligustrum lucidum Ait (LLA) in alloxan‐induced diabetic rats. OA in the alloxan‐induced diabetic rats showed significant hypoglycemic activity by lowering blood glucose (at doses of 60 and 100 mg/kg for 40 days). The levels of serum total cholesterol (TC), triglycerides (TG) and low‐density lipoprotein cholesterol (LDL‐c) in the OA‐treated diabetic rats were lower, and the high‐density lipoprotein cholesterol (HDL‐c) level was higher than in the control diabetic rats. A significant reduction in the serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP) levels of diabetic rats following OA treatment was also observed. Furthermore, OA treatment decreased the malondialdehyde (MDA) level, but increased superoxide dismutase (SOD) and glutathione peroxidase (GSH‐px) activities of the liver and kidney in diabetic rats. These results indicate that OA could protect the liver function avoiding alloxan‐induced damage; OA had hypoglycemic, hypolipidemic and antioxidant efficacy in the diabetic rats. The antioxidant ability of OA might be one of the mechanisms of its hypoglycemic and hypolipidemic effects. Copyright

Protective effects of fraction 1a of polysaccharides isolated from Solanum nigrum Linne on thymus in tumor-bearing mice.

AIM OF THE STUDY To further screen out the anti-tumor active compound of polysaccharides isolated from Solanum nigrum Linne (SNL-P), which had shown to have anti-cervical cancer and modulating properties, and evaluate the thymus protective effects of this active compound. MATERIAL AND METHODS SNL-P was separated and purified by column chromatography, and its anti-cervical cancer activity was evaluated by mice models injected of ascites U14 cells. Furthermore, the protective effect of fraction 1a of SNL-P (SNL-P1a) on the thymus tissue of tumor-bearing mice was evaluated by histological study and TUNEL staining. Finally, the protein expression of Bcl-2 and Bax gene were assayed by immunohistochemistry. RESULTS SNL-P1a has shown a marked inhibition effect on U14 cevical cancer, it restore the ratio of CD4(+)/CD8(+) peripheral blood T-lymphocyte subpopulation. Histological study and TUNEL staining results showed that SNL-P1a protect thymus tissue against the onslaught of tumor by inhibiting thymus lymphocyte apoptosis, and immunohistochemistry assay displayed that SNL-P1a treatment could increase Bcl-2/Bax ratio in thymus lymphocytes of tumor-bearer, which might promote more thymus lymphocytes towards proliferation. CONCLUSION SNL-P1a had significant growth inhibition effect on U14 cervical cancer and protective effect on thymus tissue of tumor-bearing mice.

Antitumor and immunomodulating effects of polysaccharides isolated from Solanum nigrum Linne

We have examined the effects of the crude polysaccharides isolated from Solanum nigrum Linne (SNL‐P) in vitro and in vivo against U14 cervical cancer. SNL‐P showed no antiproliferative effects in vitro at a dose up to 1 mg/ml. In vivo administration with SNL‐P (90, 180, 360 mg/kg b.w., p.o.) decreased the number of ascites tumor cells and prolonged the survival time of U14 cervical‐cancer‐bearing mice. FACScan flow cytometer analysis showed that most of the ascites tumor cells were arrested in G2/M phase of cell cycle and the ratio of CD4+/CD8+ peripheral blood T‐lymphocyte subpopulations were restored following treatment of SNL‐P. Furthermore, the treatment with SNL‐P also caused a significant increment in IFN‐γ (p < 0.01, 90, 180 and 360 mg/kg b.w.) and a remarkable decrease in Il‐4 (p < 0.01, 90, 180 mg/kg b.w.; p < 0.05, 360 mg/kg b.w.) by the method of ELISA. These data showed that SNL‐P possess potent antitumor activity and SNL‐P might exert antitumor activity via activation of different immune responses in the host rather than by directly attacking cancer cells on the U14 cervical cancer bearing mice. Thus, SNL‐P could be used as an immunomodulator and an anticancer agent. Copyright

Rhodiola sacra aqueous extract (RSAE) improves biochemical and sperm characteristics in cryopreserved boar semen.

Although Rhodiola sacra aqueous extract (RSAE) has been used in many studies as an antioxidant, its effects on semen characteristics and its antioxidant properties during cryopreservation of boar sperm have never been evaluated. Semen was collected from five Duroc boars (2-4-year-old) twice weekly and frozen-thawed in extender with RSEA. Motion characteristics were assessed with a computer-aided semen analysis (CASA) system, whereas other sperm quality end points were assessed by routine methods. The effective concentration of RSEA in extender ranged from 4 to 8mg/L and the effect of RSEA on sperm quality was better in glycerol-free extender than extender containing glycerol (P<0.05). In frozen-thawed boar semen, there was a direct correlation (P<0.05) between RSEA concentration and glutathione (GSH) concentrations, mitochondrial activity, and hypoosmotic swelling test (HOST), and an inverse correlation (r=-0.982, P<0.05) between RSEA concentration and malondialdehyde (all end points were significantly higher at 6mg/L than in the control group). In summary: (i) the effective concentration of RSEA in extender ranged from 4 to 8mg/L; (ii) the effect of RSEA on sperm quality was better in extender without glycerol; and (iii) there was a significant correlation between RSEA concentrations and concentrations of GSH and MAD in frozen-thawed boar semen (antioxidant effects of RSEA were concentration-dependent). Further studies are needed to define the active ingredient in RSEA that protects boar sperm against ROS.

Inhibition of tumor growth by recombinant adenovirus containing human lactoferrin through inducing tumor cell apoptosis in mice bearing EMT6 breast cancer

Human lactoferrin (hLTF), an 80-kDa iron-binding glycoprotein, has antitumor activity. In this study, a recombinant adenovirus containing the human lactoferrin cDNA (ad-rhLTF) was constructed and its effect on tumor growth was investigated in mice bearing EMT6 breast cancer. Ad-rhLTF was injected seven times within 14 days into the tumor site at two concentrations (108 and 5 × 108 pfu/mL) in mice bearing EMT6 breast cancer. Injected ad-rhLTF had considerable cytotoxicity on mice breast cancer, and significantly reducing the weight of tumor produced and increasing the tumor inhibition rate up to 52.64%. The presence of apoptotic cells was confirmed using TUNEL staining and flow cytometry assays. At the same time, RTPCR and Western blot analyses demonstrated that ad-rhLTF also decreased expression of Bcl-2 and increased Bax and caspase 3 expressions. Therefore, we conclude that ad-rhLTF inhibits tumor growth by inducing tumor cell apoptosis in mice with breast cancer by triggering the mitochondrial-dependent pathway and activation of caspase 3. The results indicate that ad-rhLTF might be a promising drug for breast cancer gene therapy.

Melatonin modulates the functions of porcine granulosa cells via its membrane receptor MT2 in vitro

Melatonin (N-acetyl-5-methoxytryptamine) is documented as a hormone involved in the circadian regulation of physiological and neuroendocrine function in mammals. Herein, the effects of melatonin on the functions of porcine granulosa cells in vitro were investigated. Porcine granulosa cells were cultivated with variable concentrations of melatonin (0, 0.001, 0.01, 0.1, 1.0, and 10ng/mL) for 48h. Melatonin receptor agonist (IIK7) and antagonist (Luzindole, 4P-PDOT) were used to further examine the action of melatonin. The results showed optimum cell viability and colony-forming efficiency of porcine granulosa cells at 0.01ng/mL melatonin for 48-h incubation period. The percentage of apoptotic granulosa cells was significantly reduced by 0.01 and 0.1ng/mL melatonin within the 48-h incubation period as compared with the rest of the treatments. Estradiol biosynthesis was significantly stimulated by melatonin supplementation and suppressed for the progesterone secretion; the minimum ratio of progesterone to estradiol was 1.82 in 0.01ng/mL melatonin treatment after 48h of cultivation. Moreover, the expression of BCL-2, CYP17A1, CYP19A1, SOD1, and GPX4 were up-regulated by 0.01ng/mL melatonin or combined with IIK7, but decreased for the mRNA levels of BAX, P53, and CASPASE-3, as compared with control or groups treated with Luzindole or 4P-PDOT in the presence of melatonin. In conclusion, the study demonstrated that melatonin mediated proliferation, apoptosis, and steroidogenesis in porcine granulosa cells predominantly through the activation of melatonin receptor MT2 in vitro, which provided evidence of the beneficial role of melatonin as well as its functional mechanism in porcine granulosa cells in vitro.

Antitumor activity of polysaccharides isolated from Patrinia heterophylla.

The research investigated the effect of Patrinia heterophylla Bunge (Valerianaceae) polysaccharides (PHB-P1) on U14-bearing mice. The tumor weight of mice treated with PHB-P1 (30, 60 mg/kg body weight) was significantly lower than that of the control group, a decrease of serum lactate dehydrogenase (LDH) activity was observed, and the serum alkaline phosphatase (AKP) level was increased slightly. The number of apoptotic tumor cells was significantly increased in the mice by treatment of PHB-P1 (30, 60 mg/kgbw). Cell cycle analysis showed the accumulation of tumor cells in the G2/M phase and a relative decrease of the S phase. By the immunohistochemical analysis, PHB-P1 (30, 60 mg/kgbw) might up-regulate the expression of p53 and Bax, and significantly inhibited the expression of Bcl-2 in tumor tissues. In conclusion, PHB-P1 could inhibit tumor growth and induce tumor cell apoptosis.

Anti-tumor activity of polysaccharides isolated from Patrinia scabra Bunge on U14 cervical carcinoma bearing mice.

The aim of our study was to investigate the effect of Patrinia scabra Bunge polysaccharide (PSB-P2) on cervical cancer cell (U14)-bearing mice. The tumor weight of mice treated with PSB-P2 (40, 80 mg/kg b.w.) was significantly lower than that of the control group and serum lactate dehydrogenase (LDH) activity was decreased, while serum alkaline phosphatase (AKP) level was only changed slightly. Meanwhile, the number of apoptotic tumor cells was significantly increased in the mice by the treatment of PSB-P2 (40, 80 mg/kg b.w.). At the same time, cell cycle analysis showed the accumulation of tumor cells in the G0/G1 phase and a relative decrease in the S phase. On the other hand, using the reverse transcription-polymerase chain reaction (RT-PCR) assay, PSB-P2 (40, 80 mg/kg b.w.) showed the up-regulation of p53 and Bax, and significant inhibition of Bcl-2 in tumor tissues. It suggests a possible mechanism of the inhibitory effect of PSB-P2 on tumor growth.

Effects of melatonin on follicular atresia and granulosa cell apoptosis in the porcine

The accumulation of reactive oxygen species is detrimental to the health of the ovarian follicle. The protective, antioxidant properties of melatonin, an endogenous component of porcine follicular fluid, on apoptosis of granulosa cells were evaluated in this study. Porcine granulosa cells from medium‐sized (3–5 mm), healthy follicles were cultured in serum‐free conditions with melatonin (0, 0.01, 0.1, 1.0, 10, and 100 ng/mL) with or without its receptor antagonist, luzindole, followed by evaluation of apoptotic markers in the treated cells. Results revealed that endogenous, intrafollicular melatonin concentration decreased as follicular atresia progressed, whereas the percentage of apoptotic granulosa cells increased. Spontaneous apoptosis of granulosa cells, triggered by serum deprivation in vitro, was remarkably blocked by melatonin (1.0 ng/mL melatonin, 32.7 ± 0.5%, vs. control, 47.0 ± 1.0%; P < 0.05). Treatment with 1.0 ng/mL of melatonin also significantly elevated MT2, SOD1, and GPX4 while lowering FASL, CHOP, and GRP78 mRNA abundance compared to the untreated control. The anti‐apoptotic effect and some changes of apoptotic‐relevant genes in granulosa cells invoked by melatonin supplementation were markedly blocked by luzindole, suggesting that melatonin could prevent the apoptosis of porcine granulosa cells during follicular atresia via its membrane receptors and its free‐radical‐scavenging activity. These findings provide new insights into the regulatory mechanism of melatonin in follicular atresia‐related functions. Mol. Reprod. Dev. 83: 692–700, 2016

Alkaloid from Angelicae dahuricae Inhibits HeLa Cell Growth by Inducing Apoptosis and Increasing Caspase-3 Activity

Angelicae dahuricae has been extensively used in traditional Chinese medicine not only domestically for thousands of years but also in foreign countries such as Korea, Japan, and Russia. In this study, the effect of alkaloid from Angelicae dahuricae (AAD) on inhibiting HeLa cell growth has been investigated by many approaches, including MTT, microstructure of apoptotic cells through DAPI staining and electron microscope, caspase-3 activity by chromotography, and cell cycle analysis by flow cytometry. The results showed that AAD could induce HeLa cell apoptosis, inhibit cell growth, and increase caspase-3 activity in a dose- and time-dependent manner. The mechanism of AAD-inducing apoptosis might be associated with multiple antiproliferative action towards the cells by inhibiting, preventing cell progression, and inducing a caspase-3 mediated apoptosis.