Effect of Shaoyao Tang on ulcerative colitis in rats via regulation of TLR4/NF-κB signal pathway / 中国中药杂志

The aim of this paper was to investigate the effect of Shaoyao Tang on ulcerative colitis(UC) in rats via regulation of TLR4/NF-κB signal pathway. A total of 56 Wistar rats were randomly divided into 6 groups: normal control group(double distilled water), model group(double distilled water), mesalazine group(10 mL·kg~(-1)), high dose, middle dose and low dose Shaoyao Tang groups(2.4, 1.2, and 0.6 g·mL~(-1)). After UC rat models were established by 2, 4-dinitrochlorobenzene(DNCB)/ethanol enema, the rats received double distilled water or corresponding drugs twice a day for 7 days. After the treatment cycle, the general performance and disease activity index(DAI) of rats were observed on the next day. Then the rats were sacrificed. The length of colon was measured. Macroscopic and histological score of colon were evaluated. Histopathological changes of colon were observed by HE staining. Ultraviolet spectrophotometry detection was used to detect the content of myeloperoxidase(MPO) in blood and colon tissues. The levels of P-selectin, macrophage migration inhibitory factor(MIF) and thromboxane B_2(TXB_2) in blood and colon tissues were determined by ELISA. Immunohistochemistry and Western blot analysis were performed to detect the protein expressions of TLR4 and NF-κB in colon tissues. The results showed that as compared with the model group, Shaoyao Tang of different doses improved the general performance of UC rats. Moreover, high-dose Shaoyao Tang group showed the most obvious effect in scoring of disease activity index(P<0.001); both medium and high doses of Shaoyao Tang significantly inhibited the colon shortening and pathological injury, with significantly decreased expression levels of MPO, P-selectin, MIF and TXB_(2 )in serum and colon tissues of UC rats(P<0.001). Immunohistochemistry and Western blot assay showed that the levels of TLR4 and NF-κB protein expression in the colon tissues of Shaoyao Tang high-dose group were remarkably lower than that in the model group(P<0.001). This study shows that Shaoyao Tang has protective and repairing effects on UC, and its possible mechanism is achieved probably by regulating the TLR4/NF-κB pathway and inhibiting the expressions of MPO, P-selectin, MIF and TXB_2.

Efficacy and Immune Mechanism of Baitouweng Tang on Ulcerative Colitis in Rats / 中国实验方剂学杂志

Objective: To investigate the therapeutic effect and immune mechanism of Baitouweng Tang on ulcerative colitis(UC) rats. Method: The mode of UC rats was made of 2, 4-dinitrochlorobenzene (DNCB)/ethanol enema. Rats were randomly divided into control group, model group, mesalazine group, and high-dose, middle-dose and low-dose Baitouweng Tang groups. The mesalazine group were administered with mesalazine (0.5 g·kg-1). Baitouweng Tang groups were given Baitouweng Tang (10, 5, 2.5 g·kg-1), while the other groups were given double steaming water. After 7 days of continuous administration, the general condition and disease activity index of rats in each group were observed. After anesthesia in rats, blood was taken from the abdominal aorta. Then the rats were put to death, and the length and morphological observation of the colon were measured. Ultraviolet spectrophotometry detection was used to detect the activities of myeloperoxidase (MPO) in blood and colon tissue. The levels of P-selectin, macrophage migration inhibitory factor (MIF) and thromboxane B2 (TXB2) in blood and colon tissues were detected by enzyme-linked immunosorbent assay(ELISA). Immunohistochemistry and Western blot methods were undertaken to determine the expressions of Toll-like receptor 4 (TLR4) and nuclear transcription factor-κB (NF-κB) proteins in colon tissue. Result: Compared with the model group, the rats in model group showed severe symptoms, such as loose stools, diarrhea and bloody stools, while Baitouweng Tang obviously ameliorated them. Moreover, Baitouweng Tang significantly reduced DAI, colon general and pathological scores, which were high in model group(P2 in the serum and colon tissues of the model group were obviously increased(PκB in colons of model group were markedly higher than those in control group(PPConclusion: Baitouweng Tang could inhibit TLR4/NF-κB signaling pathway in treatment of ulcerative colitis, and reduce the expressions of P-selectin, MPO, MIF and TXB2, and thus promoting intestinal mucosal repair and improving intestinal function.

Variations of sodium chloride concentration affect conformational dynamics of human neuronal calcium sensor-1 protein: a molecular dynamics simulation study / 中国组织工程研究

BACKGROUND: Human neuronal calcium sensor-1 (NCS-1) protein with a high charge is extremely sensitive to solution temperature, but whether the change of ion concentration in the solution can change the protein structure and affect the physiological function has not been reported. OBJECTIVE: To investigate the molecular mechanism of variations of sodium chloride concentration altering conformational dynamics of human NCS-1. METHODS: Through molecular dynamics simulation, the first two minimum energy models (PDB id: 2LCP) were utilized as the starting states of each independent molecular dynamics simulation of the human NCS-1 protein. After energy minimization, two different initial structures were used to conduct three independent 500 ns molecular dynamics simulations with explicit solvent for human NCS-1 protein at 0.1, 0.3 and 1 mol/L sodium chloride concentration. RESULTS AND CONCLUSION: The slightly high concentration of sodium chloride increased the flexibility of loop L2, simultaneously expanded the global and local structure of NCS-1 protein. Increase in sodium chloride concentration induced loop L3 to adopt a collapsed state and reduced the connectivity between the starting and ending residues of loop L3. N-domain and C-domain interdomain correlation was weakened and the intradomain coupling strengthened. Formation number and the probability of the salt bridges were reduced dramatically, especially at 0.3 mol/L sodium chloride. These findings manifest that human NCS-1 protein is sensitive to the small variation of sodium chloride concentration, which may alter the key factors on protein conformation. Our study may provide the theoretical reference at the atomic structural insights for probing the conformational variations of human NCS-1 protein at the different aqueous solutions.

Study of isobutyrylshikonin inhibiting proliferation of colon carcinoma cells through PI3K/Akt/m-TOR pathway / 中国中药杂志

To investigate the inhibitory effect of isobutyrylshikonin on the growth of human colon carcinoma cells and its effect on the PI3K/Akt/m-TOR pathway. MTT assay was used to detect the inhibitory effect of different concentrations (0, 6.25, 12.5, 25, 50, 100 mg·L⁻¹) of isobutyrylshikonin on the proliferation of human colon carcinoma cell HT29 at 24, 48 h. CCK-8 method was used to detect the inhibitory effect of isobutyrylshikonin on HT29, HCT116, DLD-1 and Caco-2 at 48 h. AnnexinV/propidium iodide staining was applied in detecting the apoptoticrate of HT29 cells treated with different concentrations of isobutyrylshikonin at 24 h and 48 h. Cycletest plus DNA was employed to analyze HT29 apoptosis and cell cycle after 48 h treatment with isobutyrylshikonin at different concentrations. Western blot and RT-PCR assay were used to examine the protein and mRNA expressions of PI3K, p-PI3K, Akt, p-Akt and m-TOR. The results showed that isobutyrylshikonin inhibited the proliferation of different human colon carcinoma cells, and the inhibition rate was in a dose-dependent manner. Isobutyrylshikonin induced apoptosis mainly in the early stage and blocked cells in the G₀/G₁ or G₂/M phase. Isobutyrylshikonin reduced the protein expressions of PI3K, p-PI3K, Akt, p-Akt, m-TOR and the mRNA expressions of PI3K, Akt, m-TOR in a dose-dependent manner. Isobutyrylshikonin can significantly inhibit the proliferation, induce the early apoptosis and change the cycle distribution in colon carcinoma cells.This biological effect may be correlated with the inhibition of PI3K/AKT/m-TOR pathway.

Protective effect of baicalin against LPS-induced intestinal injury / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the protective effect of baicalin on the intestinal mucosal injury caused by endotoxin-lipopolysaccharide (LPS) and the anti-oxidative injury in colonic and ileal mucosa of rats with septicopyemia.</p><p><b>METHOD</b>Fifty healthy male BALB/c mice were randomly divided into 5 groups: the normal control group, the model group, and baicalin high-dose, medium-dose and low-dose groups. They were orally administered with double distilled water, 100 mg x kg(-1) of baicalin, 50 mg x kg(-1) of baicalin, and 25 mg x kg(-1) of baicalin respectively for three days, once a day. 1 h after the oral administration on 3 d, they were intraperitoneally injected with normal saline or LPS (17 mg x kg(-1)). At 20 h after the injection of LPS, all of the mice were sacrificed, and their colonic and ileal tissues were collected. The mental status, life state and death rate of mice in each group were observed, and the lengths of colonic were measured. Chiu's scoring method was used to assess the intestinal mucosal injury. Histopathological changes of intestinal tissues were tested by HE staining. The ultraviolet spectrophotometry was used to detect total antioxidant capacity (T-AOC), superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-PX) of intestinal homogenate. The immunohistochemical method was used to analyze the expression of PCNA in intestinal tissues of each group.</p><p><b>RESULT</b>The death of mice was observed after the intraperitoneal injection of LPS. The death rates of baicalin groups were remarkably lower than the death rate of the model group. The colons in the medium-dose baicalin group were much longer than that in the model group (P < 0.05), with a much lower intestinal mucosa injury degree than the model group. Colonic and ileal injuries in the high-dose baicalin group significantly (P < 0.05). Colonic and ileal injuries in the medium-dose baicalin group and the low-dose baicalin group significantly reduced compare with the model group (P < 0.000 1). The medium-dose baicalin group showed no significant increase in homogenate's T-AOC, T-SOD and GSH-PX compare with the model group (P < 0.05). There was no significant difference between baicalin groups and the model group in PCNA.</p><p><b>CONCLUSION</b>Baicalin can protect intestinal epithelial cells suffering from injury from oxygen radicals, and relieve the intestinal injury caused by LPS by improving the intestinal mucosa structure and functions.</p>

Effect of fuzheng yiliu granule on H22 tumor cell apoptosis, p53 and caspase-3 gene expression in mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the inhibitory effect of Fuzheng Yiliu Granule (FYG) on growth and apoptosis of H22 tumor cell, and expressions of p53 and Caspase-3 gene.</p><p><b>METHODS</b>The tumor inhibitory rate of FYG on H22 tumor cell line was observed in vivo, cell apoptosis rate and cell cycle were determined by flow cytometry (FCM), and expressions of wild type p53 and Capase-3 mRNA were determined by RT-PCR.</p><p><b>RESULTS</b>FYG could inhibit the growth of H22 tumor cell at the dose of 12g/kg, 24g/kg, the maximal inhibitory rate up to 51.24% (P < 0.01). By FCM, it was shown that FYG could significantly enhance apoptosis rate of cell line H22, with the peak reached 15.84% (P < 0.01), and cause the tumor cell cycle being blocked at G0/G1 phase, with decrease of cells in S phase. RT-PCR illustrated that FYG could significantly up-regulate the level of p53 and Caspase-3 mRNA expression.</p><p><b>CONCLUSION</b>FYG can significantly inhibit the growth of tumor cell in mice, its anti-tumor mechanisms may relate to the cell apoptosis induction, cell cycle regulation and wild type p53 and capase-3 gene expression enhancing.</p>

A study on the genital system injury induced by nickel sulfate in male rats / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the mechanism of the genital system damage by nickel sulfate in male rats in order to provide the laboratory theoretical evidence for the prevention and cure of nickel genital toxicity.</p><p><b>METHODS</b>Three groups of rats were injected intraperitoneally with nickel sulfate at dose of 1.25, 2.50, 5.00 mg/kg respectively for two weeks. The content of testicle nickel and blood serum testosterone (T), follicle-stimulating hormone (FSH), luteinizing hormone (LH) were assessed with atomic absorption spectrum and radioimmuno-assay, meanwhile the activity of nitric oxide synthase (NOS) and the content of nitric oxide (NO) were measured by enzyme method.</p><p><b>RESULTS</b>The contents of testicle nickel [(0.22 +/- 0.03), (0.34 +/- 0.04), (0.41 +/- 0.02) micro g/g respectively] were increased, but the content of T, TSH, LH in blood serum were reduced; the activities of NOS in testicle tissue [(33.65 +/- 2.93), (26.53 +/- 9.52), (10.20 +/- 2.74) U/g respectively] were inhibited by nickel sulfate and the contents of NO [(0.26 +/- 0.03), (0.18 +/- 0.05), (0.15 +/- 0.02) mmol/g respectively] were decreased (P < 0.01).</p><p><b>CONCLUSION</b>Nickel-induced genital system injury of male rats may be related to the decrease in the contents of T, TSH, LH, and the inhibition on NOS, as well as the fall of NO content.</p>