Antioxidant and Antiapoptotic Polyphenols from Green Tea Extract Ameliorate CCl-Induced Acute Liver Injury in Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the phenolic composition, antioxidant properties, and hepatoprotective mechanisms of polyphenols from green tea extract (GTP) in carbon tetrachloride (CCl)-induced acute liver injury mouse model.@*METHODS@#High-performance liquid chromatography was used to analyze the chemical composition of the extract. Antioxidant activity of GTP was assessed by O, OH, DPPH, and ferric-reducing antioxidant power (FRAP) assay in vitro. Sixty Kunming mice were divided into 6 groups including control, model, low-, medium-, and high-doses GTP (200, 400, 800 mg/kg) and vitamin E (250 mg/kg) groups, 10 in each group. GTP and vitamin E were administered at a level of abovementioned doses twice per day for 7 days prior to exposure to a single injection of CCl. Hepatoprotective effects of GTP were evaluated in a CCl-induced mouse model of acute liver injury, using commercial enzyme linked immunosorbent assay kits, histopathological observation, terminal deoxynucleotidyl transferase-mediated dUTPNick-end labeling (TUNEL) assay and Western blot.@*RESULTS@#GTP contained 98.56 µg gallic acid equivalents per milligram extract total polyphenols, including epicatechingallate, epigallocatechin gallate, epicatechin, and epigallocatechin. Compared with the model group, low-, medium-, or high doses GTP significantly decreased serum levels of alanine aminotransferase and aspartate transaminase (P<0.01). Histopathological observation confirmed that pretreatment of GTP prevented swelling and necrosis in CCl-exposed hepatocytes. Hepatoprotective effects of low-, medium-, and high-dose GTP were associated with eliminating free radicals and improving superoxide dismutase, catalase, and glutathione peroxidase activity in the liver. Additionally, low-, medium-, and high-dose GTP decreased cell apoptosis in the CCl-exposed liver (P<0.01). Phosphorylated nuclear factor kappa-B (NF-κB), p53, Bcl-2 associated x protein/B-cell lymphoma/leukemia-2 gene, cytochrome C, and cleaved caspase-3 levels were downregulated compared with the model group (P<0.01).@*CONCLUSION@#GTP achieves hepatoprotective effects by improving hepatic antioxidant status and preventing cell apoptosis through caspase-3-dependent signaling pathways.

Effect of Sanhuang Yinchi Decoction in Preventing Acute Liver Injury by HMGB1 Signaling Pathway / 中国实验方剂学杂志

Objective:To investigate the effect and mechanism of Sanhuang Yinchi decoction (SHYCD) in preventing carbon tetrachloride (CCl4)-induced acute liver injury by regulating high mobility group box1(HMGB1) signaling pathway. Method:A total of 48 KM mice were randomly divided into blank control group, model group, low, middle and high-dose SHYCD groups and positive control group. The model of acute liver injury induced by CCl4 in mice was established. The low, middle and high-dose SHYCD groups were intragastrically administered with drugs (16, 32, 48 g·kg-1·d-1) respectively, and the positive control group was given cell growth stimulating hormone (20 mg·kg-1·d-1) through intraperitoneal injection. Pathological changes of mouse liver tissue sections were observed by hematoxylin-eosin staining (HE); relevant enzyme kits were used to determine liver function indexes in mice serum-alanine aminotransferase (AST) and aspartate aminotransferase (ALT); the expression level of interleukin-6 (IL-6) in mouse serum was determined by enzyme-linked immunosorbent assay (ELISA); Western blot was used to detect the expressions of high mobility group box-1(HMGB1), cysteine aspartic acid protease(Caspase-3), apoptosis-related molecules B cell lymphoma(Bcl-2), Bcl-2 associated x protein(Bax), and Toll-like receptor 4 (TLR4). Result:Compared with the normal group, the model group significantly increased serum AST, ALT (PPPPPConclusion:SHYCD can prevent liver injury by regulating HMGB1/TLR4/NF-κB signaling pathway, reducing cellular inflammatory response and inhibiting apoptosis, so as to prevent acute liver injury in mice. This indicates that HMGB1 may become a new target to prevent acute liver injury.

Pi (Spleen)-deficiency syndrome in tumor microenvironment is the pivotal pathogenesis of colorectal cancer immune escape / 中国结合医学杂志

Cancer immunoediting consists of three sequential phases: elimination, equilibrium, and escape. For colorectal adenoma-carcinoma sequence, the adenoma dysplastic progression may represent an equilibrium phase and the cancer stage as escape phase. Immune system eliminates transformed enterocytes by destroying them at first, sculpts them at the same time and selects the variants subsequently that are no longer recognized and insensitive to immune effectors, and finally induces immunosuppressive state within the tumor microenvironment that facilitates immune escape and tumor outgrowth. Immunosuppression and inflammation are the two crucial features of Pi (Spleen)-deficiency. Classic quotations, immune evidence and clinical observations suggest that Spleen (but not other organs) deficiency is the key pathogenesis of colorectal cancer (CRC) microenvironment. Weakness of old age, immunosuppressive cytokines from chronic inflammation, tumor-derived immunosuppressive factors and surrendered immune cells-regulatory T cells, myeloid-derived suppressor cells and tumor associated macrophages (TAMs) constitutes CRC microenvironment of Pi-deficiency. Furthermore, excess in superficiality, such as phlegm stagnation, blood stasis and toxin accumulation are induced by chronic inflammation on the basis of asthenia in origin, an immunosuppressive state. Great masters of Chinese medicine emphasize that strengthen Pi is the chief therapeutic principle for CRC which receives good therapeutic effects. So, Pi-deficiency based syndrome is the pivotal pathogenesis of tumor microenvironment. The immunosuppressive microenvironment facilitates immune escape which play an important role in the transition from adenoma to adenocarcinoma. There are some signs that strengthen Pi based treatment has potential capacity to ameliorate tumor environment. It might be a novel starting point to explore the mechanism of strengthen Pi based therapy in the prevention and treatment of CRC through regulation of tumor environment and immunoediting.

Protective effect of Sanhuangyinchi Fang drug serum on hydrogen peroxide-induced DNA oxidative damage in LO2 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the protective effect of Sanhuangyinchi Fang drug serum (SF) against hydrogen peroxide-mediated DNA oxidative damage in LO2 cells.</p><p><b>METHODS</b>The LO2 cells were randomly divided into the control group, H(2)O(2) group, SF groups (5%, 10%, and 15%) and vitE group. The morphological features of the treated LO2 cells were observed under inverted microscope. The viability of the treated cells was assessed with CCK-8 method, and the activity of SOD, CAT and GSH-PX were detected biochemically. Reactive oxygen species (ROS) levels, the content of 8-OHdG, and DNA damage of the cells were evaluated by flow cytometry, ELISA, and Comet assay, respectively.</p><p><b>RESULTS</b>Compared with H(2)O(2) group, the cells in SF groups (10% and 15%) and vitE group showed higher cell survival rate (P<0.05) and higher SOD, CAT, GSH-PX (P<0.05) and ROS scavenging activities (P<0.01) with markedly decreases the content of 8-OHdG (P<0.01) and reduced tailing ratio, tail length, tail moment and Olive tail moment (P<0.05).</p><p><b>CONCLUSION</b>SF drug serum, especially at the concentration of 15%, can protect LO2 cells from H(2)O(2)-mediated DNA oxidative damage.</p>

Effect of curcumin on radiosensitization of CNE-2 cells and its mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the effect of curcumin (Cur) on radiosensitivity of nasopharyngeal carcinoma cell CNE-2 and its mechanism.</p><p><b>METHOD</b>The effect of curcumin on radiosensitivity was determined by the clone formation assay. The cell survival curve was fitted by Graph prism 6. 0. The changes in cell cycle were analyzed by flow cytometry (FCM). The differential expression of long non-coding RNA was detected by gene chip technology. Part of differentially expressed genes was verified by Real-time PCR.</p><p><b>RESULT</b>After 10 micro mol L-1 Cur had worked for 24 h, its sensitization enhancement ratio was 1. 03, indicating that low concentration of curcumin could increase the radiosensitivity of nasopharyngeal carcinoma cells; FCM displayed a significant increase of G2 phase cells and significant decrease of S phase cells in the Cur combined radiation group. In the Cur group, the GUCY2GP, H2BFXP, LINC00623 IncRNA were significantly up-regulated and ZRANB2-AS2 LOC100506835, FLJ36000 IncRNA were significantly down-regulated.</p><p><b>CONCLUSION</b>Cur has radiosensitizing effect on human nasopharyngeal carcinoma CNE-2 cells. Its mechanism may be related to the changes in the cell cycle distribution and the expression of long non-coding IncRNA.</p>

Effect of Lianggesan on the expression of signal transducer and activator of transcription 1 in a rat model of lipopolysaccharide-induced acute lung injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Lianggesan on the expression of signal transducer and activator of transcription 1 (STAT1) in rats with lipopolysaccharide (LPS)-induced acute lung injury and explore the possible mechanisms of the therapeutic effects.</p><p><b>METHODS</b>Endotoxemia was induced in Wistar rats by intravenous injection of LPS (5 mg/kg). The rats were randomly divided into 6 groups, namely the control group, acute lung injury group (LPS group), 3 Lianggesan groups treated at different doses, and LPS+DEX treatment group. Each group, except for the control group, was further divided into 5 subgroups and examined at 1, 2, 4, 8 and 16 h after LPS injection. Western blotting was used to detect the protein expression of STAT1 and p-STAT1 in the lung tissue.</p><p><b>RESULTS</b>In LPS group, the expression of STAT1 began to increase at 1 h following LPS injection, reaching the peak level at 4 h; the peak expression of p-STAT1 occurred at 2 h after LPS administration (P<0.01). Compared with LPS group, the 3 Lianggesan groups and DEX group showed significantly decreased expressions of STAT1 and p-STAT1 at 2, 4 and 8 h after LPS injection (P<0.05 or 0.01).</p><p><b>CONCLUSION</b>Abnormal expression of STAT1 occurs in the lung tissue in the event of ALI. Lianggesan can relieve LPS-induced acute lung injury in rats by decreasing the expression of STAT1 and p-STAT1.</p>

High-performance liquid chromatography for determining celecoxib concentration in hamster tongue tissue after oral local application / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a high-performance liquid chromatography (HPLC)-based method for determining celecoxib concentration in the tongue tissue of hamsters.</p><p><b>METHODS</b>Celecoxib mixed with the matrix (final concentration of 6%) was smeared on the surface of the tongue mucosa of hamsters, and the concentration and absorption rate of celecoxib in the tongue tissue were determined by HPLC at 5, 10, 15, 30, 60, 90, 120 min after the application.</p><p><b>RESULTS</b>In this system, the retention time of celecoxib was 4.4 min. Celecoxib concentration showed a good linear range within 25-800 microg/L (R2=0.9991, n=6), with the detection limit for celecoxib of 10 g/L (S/N=3). The extraction recoveries and method recoveries for celecoxib were 83.75%-90.01% and 91.98%-99.07%, respectively. The inter-day RSDs were 2.15%, 3.16% and 3.67%, and intra-day RSDs were 3.40%, 4.56% and 4.42%, respectively. The concentration of celecoxib in hamster tongue tissue within the first 120 min ranged from 0.685-/+0.019 microg/g to 3.168-/+0.143 g/g, reaching the peak level at 15 min.</p><p><b>CONCLUSION</b>Celecoxib can be rapidly absorbed through the tongue mucosa to reach a high concentration in the tongue tissue, indicating the possibility of oral COX-2 inhibitors to prevent oral cancer and precancerous lesions.</p>

Molecular mechanism of Wulongdan for improving the learning and memory abilities of rats with chronic cerebral ischemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of Wulongdan on the learning and memory abilities of rats with chronic cerebral ischemia and explore the mechanisms.</p><p><b>METHODS</b>Male SD Rat models of chronic cerebral ischemia were established by permanent ligation of the bilateral carotid arteries. Three weeks after the operation, the rats were randomly divided into sham-operated group, chronic cerebral ischemia group (model group), high-dose drug group, low-dose drug group and Yinxingye group and received the corresponding treatments on a daily basis for 5 consecutive weeks. Morris water maze was used to assess the learning and memory abilities of the rats, and Western blotting was carried out for detecting the expressions of NR1 and NR2B in the hippocampus and cerebral cortex.</p><p><b>RESULTS</b>Compared with the model group, the rats in high-dose drug, low-dose drug and Yinxingye groups showed significantly shorter time of finding platform in Morris water maze test (P<0.05 or 0.01). The rats in the model group showed significantly lowered expressions of NR1 and NR2B of the cortex and hippocampus than those in the sham-operated group (P<0.01). In comparison with the model group, the high-dose Wulongdan group and Yinxingye group both showed significantly increase expressions of NR1 and NR2B (P<0.01), but their expression levels still remained significantly lower than those in the sham-operated group (P<0.01).</p><p><b>CONCLUSION</b>Wulongdan can enhance the learning and memory abilities of rats with chronic cerebral ischemia, the mechanisms of which may involve increased expressions of NR1 and NR2B in the hippocampus and cortex.</p>

Effect of Oviductus Ranae on Cyclin D1, CDK6 and P15 expressions in the liver tissue of aged male rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of Oviductus Ranae (OR) on the expressions of CyclinD1, CDK6 and P15 in the liver of aged male rats.</p><p><b>METHODS</b>Eighteen male SD rats were randomly divided into 3 equal groups, namely the OR group, VE group and ageing model group. The rats received subcutaneous injection of D-galactose for 6 weeks to establish the aging models, and another 6 rats were injected daily with normal saline (NS) to serve as the normal control group. From the third week of the experiment, the rats were given oral OR or Vitamin E (VE) accordingly till the sixth week. After completion of the drug administration, all the rats were sacrificed for detecting the expressions of CyclinD1, CDK6 and P15 in the liver tissue by Western blotting.</p><p><b>RESULTS</b>The relative expression levels of CyclinD1, CDK6 and P15 in the liver of the rats in the OR group were 41.73-/+0.54, 23.29-/+0.30 and 1.49-/+0.30, respectively, significantly up-regulated as compared with those in the ageing model group (P<0.01). The expressions of the proteins were obviously down-regulated in the model group in comparison with those in the normal control group.</p><p><b>CONCLUSIONS</b>OR treatment can lower the expressions of Cyclin D1 and CDK6 in the liver to enhance the liver cell proliferation in aged male rats. OR also promotes the expression of P15 through a feedback mechanism to prevent excessive proliferation of the cells. The effect of OR against ageing is mediated possibly by up-regulation of the proteins associated with the cell proliferation in the liver, a mechanism different from that of VE.</p>

Expression of extracellular signal-regulated protein kinases in the subcutaneous fascia of rats and their changes after acupuncture / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of acupuncture on the expression of extracellular signal-regulated protein kinases 1/2 (ERK1/2) in the subcutaneous fascia of SD rats.</p><p><b>METHODS</b>Eighteen SD rats were randomly divided into 6 groups (n=3) including 5 acupuncture groups and a control group. The rats in the 5 acupuncture groups received electro-acupuncture therapy in the regions of the inguinal groove, and at 0, 1, 6, 12, and 36 h after the last therapy, the superfacial fascia surrounding the acupuncture point (about 1.5 cm in diameter) were collected. The fascia tissues at the corresponding sites and at the acupoint Zusanli (ST36) were obtained from the control rats. The expression of ERK1/2 and phosphorylated ERK1/2 (p-ERK1/2) in the tissues were detected by Western blotting.</p><p><b>RESULTS</b>ERK1/2 and p-ERK1/2 expressions were detected in the tissues harvested from both the acupoint and the non-acupoint in the control rats with similar expression intensities. In the rats of each acupuncture group, ERK1/2 expression was significantly increased on the acupuncture side in comparison with the control side.</p><p><b>CONCLUSION</b>The normal loose connective tissue may participate in tissue proliferation and differentiation possibly via phosphorylation of ERK. Acupuncture can promote the signal transduction pathway of ERK, which can be a possible mechanism for the effect of acupuncture in modulating the physiopathological conditions.</p>