Endoplasmic reticulum stress induces apoptosis of arginine vasopressin neurons in central diabetes insipidus via PI3K/Akt pathway.

AIMS: Central diabetes insipidus (CDI), a typical complication caused by pituitary stalk injury, often occurs after surgery, trauma, or tumor compression around hypothalamic structures such as the pituitary stalk and optic chiasma. CDI is linked to decreased arginine vasopressin (AVP) neurons in the hypothalamic supraoptic nucleus and paraventricular nucleus, along with a deficit in circulating AVP and oxytocin. However, little has been elucidated about the changes in AVP neurons in CDI. Hence, our study was designed to understand the role of several pathophysiologic changes such as endoplasmic reticulum (ER) stress and apoptosis of AVP neurons in CDI. METHODS: In a novel pituitary stalk electric lesion (PEL) model to mimic CDI, immunofluorescence and immunoblotting were used to understand the underlying regulatory mechanisms. RESULTS: We reported that in CDI condition, generated by PEL, ER stress induced apoptosis of AVP neurons via activation of the PI3K/Akt and ERK pathways. Furthermore, application of N-acetylcysteine protected hypothalamic AVP neurons from ER stress-induced apoptosis through blocking the PI3K/Akt and ERK pathways. CONCLUSION: Our findings showed that AVP neurons underwent apoptosis induced by ER stress, and ER stress might play a vital role in CDI condition through the PI3K/Akt and ERK pathways.

Vascular-targeted TNFα improves tumor blood vessel function and enhances antitumor immunity and chemotherapy in colorectal cancer.

Delivery and penetration of chemotherapeutic drugs into neoplasm through the tumor vasculature are essential mechanisms to enhance the efficiency of chemotherapy. "Vascular targeting" strategy focuses on promoting the infiltration of chemotherapeutic drugs into neoplastic tissues. In this study, we achieved a targeted therapy by coupling tumor necrosis factor α (TNFα) with TCP-1, a novel vascular-targeting peptide, in an orthotopic colorectal cancer model in mice. High dose of TCP-1-conjugated TNFα (TCP-1/TNFα: 5µg/mouse) displayed potent antitumor activity by inducing apoptosis and reducing microvessel number in tumors than unconjugated TNFα, with no evidence of increased toxicity. In the combined therapy, the antitumor action of 5-fluorouracil (5-FU) was potentiated when the mice were pretreated with a low dose of TNFα (1ng/mouse) and to a greater extent by the same concentration of TCP-1/TNFα. In this regard, TCP-1/TNFα combined with 5-FU synergistically inhibited the tumor growth, induced apoptosis and reduced cell proliferation. More importantly, TCP-1/TNFα normalized the tumor vasculature and facilitated the infiltration of immune cells to neoplasm as well as attenuated the immunosuppressing effects of TNFα in bone marrow and spleen. At the same time, TCP-1/TNFα significantly improved 5-FU absorption into the tumor mass. Taken together, these findings underscore the therapeutic potential of TCP-1 as a drug carrier in cancer therapy. TCP-1 is a novel vascular-targeting peptide and appears to be a promising agent for drug delivery. TCP-1 fused with TNFα holds great promise for colorectal cancer therapy.

Dingxin recipe ( ) prevents ischemia/reperfusion-induced arrhythmias via up-regulating prohibitin and suppressing inflammatory responses.

OBJECTIVE: To identify the underlying mechanisms of the protective effects of Dingxin Recipe (: , DXR), a Chinese compound prescription that has been used clinically in China for more than 20 years, on ischemia/reperfusion (I/R)-induced arrhythmias in rat model. METHODS: A total of 30 rats were randomly divided into three groups: sham group, I/R group, and DXR-pretreated I/R (DXR-I/R) group. Rats in the DXR-DXRI/R group were intragastrically administrated with DXR (12.5 g/kg per day) for consecutive 7 days, while rats I/in the sham and I/R groups were administrated with normal saline. Arrhythmias were introduced by I/R and electrocardiograms (ECG) were recorded. Two-dimensional (2-D) polyacrylamide gel electrophoresis and matrix-matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) were used to identify assisted differentially expressed proteins. Immunohistochemistry, real-time quantitative polymerase chain reaction (RQ-RQPCR), Western blot, and enzyme-linked immunosorbent assay (ELISA) were performed to analyze proteins PCR), obtained in the above experiments. RESULTS: DXR significantly reduced the incidence and mean duration of ventricular tachycardia and ventricular fibrillation and dramatically decreased the mortality, as well as arrhythmia score, compared with those of the I/R group. Among successfully identified proteins, prohibitin (PHB) and heart fatty acid binding protein (hFABP) were up-regulated in DXR-pretreated I/R rats compared with those of the I/R rats. In addition, compared with the I/R group, the level of glutathione (GSH) was elevated accompanied by reduced expressions of interleukin-6 (IL-6) and neutrophil infiltration in I/R rats with DXR pretreatment. CONCLUSIONS: DXR could alleviate I/R-induced arrhythmias, which might be related to increased expression of PHB. The enhanced expression of PHB prevented against the depletion of GSH and consequently inhibited apoptosis of cardiomyocytes. Furthermore, up-regulation of PHB might ameliorate I/R-induced cell death and leakage of hFABP by suppressing neutrophil infiltration and IL-6 expressions.

Proteomic analysis of protective effects of polysaccharides from Salvia miltiorrhiza against immunological liver injury in mice.

This study was designed to investigate mechanisms of the protective effects of Salvia miltiorrhiza polysaccharide (SMPS) against lipopolysaccharide (LPS)-induced immunological liver injury (ILI) in Bacille Calmette-Guérin (BCG)-primed mice. Two-dimensional difference gel electrophoresis (2D-DIGE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis showed that three proteins are down-regulated and six proteins are up-regulated by SMPS. SMPS reduces the degree of liver injury by up-regulating the enzymes of the citric acid cycle, namely malate dehydrogenase (MDH) and 2-oxoglutarate dehydrogenase complex. LPS significantly increases nuclear factor kappa B (NF-κB) activation, inducible nitric oxide synthase (iNOS) expression and MDA level in BCG primed mice liver, whereas SMPS treatment protects against the immunological liver injury through inhibition of the NF-κB activation by up-regulation of PRDX6 and the subsequent attenuation of lipid peroxidation, iNOS expression and inflammation.

[Effect of Sanhuangyinchi decoction on liver damage and caspase-3 in rats with acute hepatic failure].

OBJECTIVE: To investigate effect of Sanhuangyinchi decoction (SHYCD) on liver damage and the pro-apoptotic factor caspase-3 in rats with acute hepatic failure. METHODS: SD rats were randomly divided into blank control group, model group, Angongniuhuang group (AGNH) and SHYCD group. Acute hepatic failure was induced in the rats by intraperitoneal injections of D-GaLN and LPS, and the death rate, ALT, TBIL, PT and caspase-3 activity was observed or tested. RESULTS: At 36 h after the injections, the death rate of the rats was 74.29% (26/35) in the model group, 31.43% (11/35) in AGNH group and 28.57%(10/35) in SHYCD group. The death rate, ALT, TBIL, PT and caspase-3 activity in AGNH and SHYCD groups were significantly lower than those in the model group (P<0.01). SHYCD showed stronger effect than AGNH in depressing TBIL and the activity of caspase-3 (P<0.05). CONCLUSION: SHYCD can improve the liver function and inhibit the activity of caspase-3 in rats, which can be the possible mechanism for its therapeutic effect against acute hepatic failure.

Effects of dahuangzhechong pills on cytokines and mitogen activated protein kinase activation in rats with hepatic fibrosis.

UNLABELLED: RELEVANCE TO ETHNOPHARMACOLOGY: Dahuangzhechong pill (DHZCP), a well-known and canonical Chinese medicine formula from "The Synopsis of Prescriptions of the Golden Chamber", is officially approved and recommended by Chinese association of integrative medicine for the prevention and treatment of hepatic fibrosis in China. AIM OF THE STUDY: To test the hypothesis that therapeutic effects of DHZCP on hepatic fibrosis are conferred by regulating cytokine profile through a mitogen activated protein kinase (MAPK) pathway. MATERIALS AND METHODS: Hepatic fibrosis is inducted by carbon tetrachloride (CCl(4)) in rats which then were randomly divided into six groups: hepatic fibrosis model group, high dose DHZCP group, low dose DHZCP group, Fufang Biejia Ruangan Pian (FBRP) group, Colchicine group and control group. Pathological, immunohistochemical, multiplex immunoassay and protein expression studies (Western blotting) are performed. RESULTS: DHZCP significantly decreases the levels of alanine aminotransferase, aspartate aminotransferase, hyaluronic acid, laminin, type IV collagen and procollagen III, and reverses hepatic fibrosis in rat model. DHZCP also could reduce the expression of α-smooth muscle actin, and lower the serum level of tumor necrosis factor alpha (TNF-α) and interleukin 13 (IL-13). The expressions of phosphorylated p38 MAPK and extracellular signal-regulated kinase (ERK) are down-regulated, while no significant changes are found in phosphorylation of c-Jun N-terminal kinase (JNK). CONCLUSIONS: DHZCP can alleviate hepatic fibrosis induced by CCl(4). The anti-fibrotic effects of DHZCP are conferred by decreasing the secretion of TNF-α and IL-13 through down-regulating p38 and ERK phosphorylation.

[Prophylactic immunization of dangguibuxue decoction against Cryptosporidium infection in immune suppressed mice].

OBJECTIVE: To explore the prophylaxis of dangguibuxue decoction, a traditional Chinese medicine made from Angelica sinensis and Radix astragalus, on immunosuppressed mice infected by Cryptosporidium parvum. METHODS: 48 BALB/c mice were randomly divided into 4 groups: normal control (A), immunosuppressed control (B), high dose (C), and low dose (D). Mice in groups B, C and D were intragastrically administered with dexamethasone (DXM) for 8 days, and in the same time mice in groups C and D were given high dose (2 g/kg) and low dose (1 g/kg) dangguibuxue decoction respectively. On the ninth day all mice in groups B, C and D were orally inoculated by 1 x 10(6) oocysts of C. parvum. The amount of oocysts in feces was examined daily since being infected. 11 days after infection, the subset of T lymphocytes in peripheral blood was analyzed with flow cytometry, sIL-2R in serum and sIgA of intestinal fluid were detected by ELISA. Pathological change of duodenum and jejunum was observed microscopically. RESULTS: Compared with the immuno-suppressed control group, there were less oocysts in feces (35.0 +/- 4.21) (P < 0.01) and lighter injury in the intestinal mucosa in mice of the high dose dangguibuxue decoction group. Both the number of CD4+ T lymphocytes (47.483 +/- 4.082) and the ratio of CD4+/CD8+ (2.271 +/- 0.378) increased, sIgA [(320.19 +/- 1.94) ng/ml] in the intestinal fluid elevated and sIL-2R [(321.34 +/- 6.66) ng/ml] in peripheral blood decreased in the high dose group, with a significant difference in comparison to the immunosuppressed group (P < 0.01). All the above-mentioned indices in low dose dangguibuxue decoction group showed no significant difference with the immunosuppressed control group (P > 0.05). CONCLUSION: Administration of high dose dangguibuxue decoction plays a role of prophylaxis on the infection of C. parvum in immunosuppressed mice through improving the immune status.