Icariin Inhibits Proliferation of CLC5 Hepatoma Cells via Akt/GSK3β/CDK Pathway / 中国实验方剂学杂志

ObjectiveTo study the effect of icariin on the proliferative capacity of hepatocellular carcinoma cell line CLC5 and the underlying mechanism. MethodThe targets of icariin were screened out by network pharmacology， and the target network and protein-protein interaction （PPI） network were constructed to predict the possible targets and pathways of icariin. CCK-8 assay was employed to explore the effects of different concentrations （0， 6.25， 12.5， 25， 50 μmol·L-1） of icariin on the viability of CLC5 cells. Further， CLC5 cells were treated with 0， 25， 50 μmol·L-1 icariin， and the effect of icariin on CLC5 cell proliferation was examined by Edu-488 assay and clone formation assay （CFA）. Western blot was employed to measure the expression levels of proteins in the protein kinase B （Akt）/glycogen synthase kinase 3β （GSK3β）/cell cycle-dependent kinase （CDK） pathway in the CLC5 cells exposed to different concentrations of icariin. ResultNetwork pharmacological analysis revealed that icariin may inhibit the hepatocellular carcinoma via cell cycle arrest and inhibition of tumor cell proliferation. Compared with the blank group， icariin decreased the viability of CLC5 cells in a time- and concentration-dependent manner （P<0.01） and reduced the positive rate of Edu-488 and the colonies in CFA （P<0.05， P<0.01）. Moreover， icariin down-regulated the protein levels of p-Akt， p-GSK3β， CDK4， and CyclinD1 （P<0.05， P<0.01）. ConclusionIcariin may block cell cycle to suppress the proliferation of CLC5 cells via inhibiting the Akt/GSK3β/CDK pathway.

Research on HBV DNA inhibition of plasmid acute infection mouse with betulinic acid / 中国中药杂志

Betulinic acid is a naturally occurring pentacyclic triterpenoid, which has antiretroviral, antimalarial, and anti-inflammatory properties. The purpose of this study is to investigate the HBV DNA replication inhibition in the mouse model with betulinic acid. Hydrodynamic injection method via the tail vein with the Paywl. 3 plasmid was used to establish the animal mode (n = 15), and the mice were randomly divided into the PBS control group (n = 5), Betulinic acid treatment group (n = 5) and lamivudine control group (n = 5). The day after successful modeling , the mice would have taken Betulinic acid (100 mg x kg(-1)), lamivudine (50 mg x kg(-1)), PBS drugs orally, once daily for 7 days, blood samples were acquired from the orbital venous blood at 3, 5, 7 days after the administering, HBsAg and HBeAg in serum concentration were measured by ELISA and the mice were sacrificed after 7 days, HBV DNA southern detections were used with part of mice livers. The results showed that betulinic acid significantly inhibited the expression of HbsAg in the mice model at the fifth day compared with the control group, and there was no significant differences between the effects of lamivudine and the PBS control group; both the betulinic acid and lamivudine groups had no significant inhibition for the HBeAg expression; the HBV DNA expressions of the liver tissue from the betulinic acid and lamivudine groups were inhibited compared with the control group. Taken together, these results reveal betulinic acid can inhibit the HBsAg expression and replication of the liver HBV DNA in the mouse model.

Characteristic of liver pathology in HBeAg-positive and HBeAg-negative chronic hepatitis B patients with mildly elevated ALT / 中华肝脏病杂志

<p><b>UNLABELLED</b>To analyse the live pathology characteristics in mild ALT-elevated (1 x ULN less than ALT less than 2 x ULN ) HBeAg-positive and HBeAg-negative chronic hepatitis B (CHB) patients, and to explore the influence of the age and HBV DNA level to liver pathology in different HBeAg status patients.</p><p><b>METHODS</b>All the patients who met the inclusion criteria form "eleventh five-year plan" National Science and Technology Major Project, the treatment program of integrative traditional and western medicine for CHB were enrolled in this study between October 2009 and March 2011 .B type ultrasound-guided liver biopsy was carried out in all patients and hepatitis B surface antigen (HBsAg) , HBeAg titer as well as HBV DNA level were detected at the same time. Hepatic tissue inflammation and fibrosis degree of patients according to HBeAg-positive and negative, age ( more than or equal to 40 years and less than 40 years), HBV DNA level (more than or equal to 10

Lingmao recipe inhibits starvation induced autophagy in HepG2.2.15 cell / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effects of Lingmao Recipe (LR) on starvation-induced autophagy in HepG 2.2.15 cell.</p><p><b>METHODS</b>Fifteen SD rats were selected to prepare LR drug serum. The Earle's balanced salt solution (EBSS) group, the EBSS + vehicle serum group, the EBSS + 5 times LR serum group, the EBSS + 10 times LR serum group, and the DMEM group were set up, 3 samples in each group. The HepG2.2.15 cells were cultured for 1, 2, 4, and 6 h respectively. The pEGFP-N1-LC3B eukaryotic expression vector was constructed and transfected with HepG2. 2.15 cell. The GFP-LC3B morphological changes were observed under fluorescent microscope. The ratio of the dotty GFP-LC3B number and the GFP-LC3B transfected HepG2.2.15 number was calculated. The intracytoplasmic autophagosome changes were observed using electronic transmission electron. The microtubule-associated protein 1 light chain 3 beta II/I was detected in HepG2.2.15 of each group using Western blot.</p><p><b>RESULTS</b>Two h after culture, when compared with the EBSS + vehicle serum group, GFP-LC3B changing from diffused distribution to dotted distribution was obviously inhibited in the EBSS +5 times LR serum group and the EBSS +10 times LR serum group (P<0.01). The electronic transmission electron showed that the formation of autophagosome was inhibited in the EBSS +5 times LR serum group and the EBSS +10 times LR serum group. Results of Western blot showed that microtubule-associated protein 1 light chain 3 beta II/I obviously decreased more in the EBSS +10 times LR serum group than in the EBSS +vehicle serum group (P<0.01).</p><p><b>CONCLUSION</b>LR could inhibit starvation-induced autophagy in HepG 2.2.15.</p>

Epidemic survey on TCM syndrome typing and preliminary discussion on TCM etiology and pathogenesis of fatty liver / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the common TCM syndrome types of fatty liver by way of epidemic questionnaire, their occurrence ratio, and the correlation between various syndrome types and objective indexes.</p><p><b>METHODS</b>A total of national wide 503 subjects with fatty liver were enrolled, the TCM syndromes, body mass index (BMI), abdominal perimeter/hip circumference, liver function, blood lipids, B ultrasonic examination and CT in them were observed and recorded.</p><p><b>RESULTS</b>In the 46 symptoms investigated in total, the first ten symptoms in order of appearing rate were lassitude, obese, oral dryness, vertigo, hypochondriac distending pain, soreness and pain in loin, spiritlessness, oral bitterness, aching and weakness in knee and abdominal distention. The mostly appeared tongue figures were pale and corpulent or pale dim tongue proper, white greasy or yellow greasy tongue coating, and the mostly appeared pulse figures were taut, taut-thin and taut slippery. Statistical cluster analysis showed that syndromes of fatty liver could be typed into 4 TCM types, the asthenia Pi-Shen with Gan-stagnation type, the asthenia Pi-Shen type, the asthenia Pi with phlegm-heat type and the unclassified type. Among them the asthenia Pi-Shen with Gan-stagnation type was the commonest one, which accounted to 62.32%.</p><p><b>CONCLUSION</b>The mostly appeared syndrome type of fatty liver was asthenia Pi-Shen with Gan-stagnation type. The TCM pathogenesis of fatty liver was deficiency of origin, mainly deficiency of Shen, involving Pi, with excess superficiality, the turbid-phlegm and blood stasis.</p>

Effect of Bushen recipe on T-cell subsets and their function in patients with chronic hepatitis B / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the regulatory effect of Bushen recipe (BSR) on immune function in patients with chronic hepatitis B.</p><p><b>METHODS</b>Thirty patients with HBeAg+ and abnormal alanine transaminase (ALT) level were treated with BSR for 6 months. The following parameters before treatment, 3 months and 6 months after treatment were tested: (1) HBV DNA by fluorescence quanitiative PCR; (2) Levels of interleukin-10 (IL-10) and interferon-gamma in supernatant of cultured peripheral blood mononuclear cell (PBMC) after had been cultured with HBeAg, HBcAg and phytohemagglutinin (PHA) for 48 hrs; (3) Blood levels of T-cells, including CD4+ , CD8+ , CD8+ CD28+ , CD8+ CD28- and CD28+ , and their ratio, as well as the percentage of CD8+ CD28+ in the cell culture after being co-cultured with PHA for 48 hrs.</p><p><b>RESULTS</b>In patients effectively treated by BSR, after 3 months treatment, level of IFN-gamma in PBMC culture significantly increased, CD8+ CD28+ , CD28+ T-cells significantly enhanced and IL-10 and CD8 + CD28+ T-cells significantly decreased (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>BSR could significantly strengthen the expression of Th1 type cytokine, decrease the expression of Th2 type cytokine and promote the expression of CD8+ cytotoxic T-lymphocyte, which may be one of mechanisms in suppressing HBV DNA replication.</p>