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Tubiechong (Eupolyphaga sinensis) is an important material used in traditional Chinese medicine (TCM). However, the immunoregulation effects of E. sinensis Lyophilized Powder (ESL) are uncl...

Huoxuezhitong capsule (HXZT, activating blood circulation and relieving pain capsule), has been applied for osteoarthritis since 1974. It consists of Angelica sinensis (Oliv.) Diels, Panax notoginseng (Burkill) F. H. Chen ex C. H., Boswellia sacra, Borneol, Eupolyphaga sinensis Walker, Pyritum. However, the direct effects of HXZT on osteoarthritis and the underlying mechanisms were poorly understood. In this study, we aimed to explore the analgesia effect of HXZT on MIA-induced osteoarthritis rat and the underlying mechanisms. The analgesia and anti-inflammatory effect of HXZT on osteoarthritis in vivo were tested by the arthritis model rats induced by monosodium iodoacetate (MIA).. Mechanistic studies confirmed that HXZT could inhibit the activation of NF-κB and down-regulate the mRNA expression of related inflammatory factors in LPS-induced RAW264.7 and ATDC5 cells. Furtherly, in LPS-induced RAW264.7 cells, HXZT could suppress NF-κB via inhibiting PI3K/Akt pathway. Taken together, HXZT capsule could ameliorate MIA-induced osteoarthritis of rats through suppressing PI3K/ Akt/ NF-κB pathway.

The traditional medicine-Eupolyphaga sinensis walker is a rich source of functional proteins and peptides. In this study, Eupolyphaga sinensis walker were identified to include fifteen amino acids, of which hydrophobic amino acids accounted for 46.7%. Eupolyphaga sinensis walker polypeptides (EPs) were extracted by sequentially hydrolyzing with pepsin and trypsin. EPs could effectively scavenge 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·), superoxide anion radical (O2−·), and hydroxyl radical (OH-·), and reduce ferric solution in vitro. EPs also enhanced the activities of Ssuperoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPH-Px), increased the contents of hydroxyproline (HYP), and reduced the content of Malondialdehyde (MDA) in photoaged mice skin. Histological study confirmed that EPs improved UV irradiation-induced damage of skin texture and morphology. Therefore, the extracted EPs are effective antioxidants and can serve as powerful treatment for skin photoaging.

Aim of the study. To examine the antiproliferation and anti-invasion of Eupolyphaga sinensis Walker 70% ethanol extract (ESWE) on breast cancer and elucidate the underlying signaling mechanisms. Methods. MTT and colony formation assays were used to investigate the effect of ESWE on proliferation of breast cancer cells in vitro. The xenograft mouse tumor model was used to determine the effect of ESWE on breast cancer in vivo. To investigate the underlying molecular mechanisms, we used western blotting to analyze the expression of ERK1/2, CXCR4, matrix metalloproteinase 2 (MMP2), and MMP9 pretreated with ESWE. The stromal cell–derived factor (SDF)-1α-induced migration and invasion potential of breast cancer cells were examined by wound-healing assays and Matrigel invasion chamber assays. Results. ESWE effectively inhibited the proliferation of MDA-MB-435s and MDA-MB-231 cells and exhibited antitumor effects in an MDA-MB-231 xenograft mice model. Furthermore, ESWE suppressed the activity of ERK1/2, a key molecule of MAPK signaling. We also observed that ESWE treatment led to downregulation of CXCR4 expression as well as greatly reduced MMP2 and MMP9. ESWE affected CXCR4 expression partially through the modulation of autocrine vascular endothelial growth factor. However, suppression of CXCR4 expression was the result of downregulation of mRNA expression. Inhibition of CXCR4 expression by ESWE further correlated with the suppression of SDF-1α-induced migration and invasion in breast cancer cells. Conclusion. ESWE exerted its antiproliferation and antiinvasion by regulating MAPK signaling and related metastasis factorsand thus could be a useful therapeutic candidate for breast cancer intervention.

An insect trypsin-like enzyme with similar biochemical properties to psychrophilic analogs was purified from Chinese ground beetle (Eupolyphaga sinensis) for the first time. The purified trypsin-like enzyme was designated as E. sinensis trypsin-like enzyme (ESTL) with molecular weight of 22.8 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The maximal activity of ESTL was observed at pH 9.5, and the temperature optimum of ESTL was observed at 45C by its amidolytic effect on the substrate benzoyl-L-Arg-p-nitroanilide. The analysis of inhibitors showed that specific inhibitor of serine proteases (phenylmethanesulfonyl fluoride) and trypsin inhibitors (tosyl–lysine chloromethyl ketone, bovine pancreatic trypsin inhibitor and benzamidine) inactivated ESTL almost completely. Over the range of tested temperature of 10–30C, the catalytic efficiency (kcat/Km) of the ESTL was about five times than that of bovine trypsin. The N-terminal sequencing of ESTL revealed the following sequence: I1VGGSTTTIQ10NFPYQVSL, and the complementary DNA (cDNA) of ESTL was cloned by rapid amplification of cDNA ends. Practical Applications In recent years, increasing attention has been drawn to the psychrophilic (cold-adapted) enzymes. Psychrophilic trypsins with high activity may be interesting for several industrial applications of enzymes, such as in certain food-processing operations that require low-processing temperatures. For example, cod trypsin is already used in food production and cosmetics. In this article, a trypsin-like enzyme [Eupolyphaga sinensis trypsin-like enzyme (ESTL) ] was purified from Chinese ground beetle by a relatively simple method, and the complementary DNA (cDNA) of this enzyme was cloned by rapid amplification of cDNA ends. This enzyme was characterized for molecular and enzymatic properties, as demonstrated that it had high activity at low and moderate temperature compared with the mammalian trypsins, and maintained good thermal stability compared with psychrophilic trypsins. ESTL has potential interest in food industry.

The insect of Eupolyphaga sinensis Walker has been used as traditional anti-thrombosis medicine without bleeding risk for several hundreds years in eastern countries. Our previous work has identified a bi-functional anti-thrombosis protein containing both direct-acting fibrin(ogen)olytic and plasminogen-activating activities from the insect. By proteomics and transcriptome analysis, 105 serine proteases belonging to four families were identified from the ground beetle, E. sinensis and the classification is for serine proteases of this organism. Pharmacological test indicated that 5 (eupolytin 1-5) of them have the abilities to hydrolyze fibrin(ogen) and/or activate plasminogen. The current work revealed the extreme diversity of anti-thrombosis components in E. sinensis and anti-thrombosis molecular mechanisms of the traditional medicinal insect, and provided many templates for the development of new thrombolytic agents. Especially, these proteins, which contain both plasmin- and PA (plasminogen-activating)-like activities, are excellent candidates for anti-thrombosis medicines.

Ethnopharmacological relevance Eupolyphaga sinensis Walker popularly known as “preferred drug to regulate blood flow” are traditionally used in folk medicine in the treatment of ecchymoma, posttraumatic wound, hepatic fibrosis and tumor. Aim of the study To characterize chemical compositions and to evaluate the antitumor and immunomodulatory of Eupolyphaga sinensis Walker ethanol extract (ESEE) in hepatocarcinoma H22 bearing mice. Materials and methods ESEE was obtained by ethanol reflux extraction and analyzed by gas chromatography–mass spectrometry (GC–MS) after methylation. ICR mice were treated with ESEE for 14 consecutive days at doses of 31 mg/kg (low-dose), 62 mg/kg (mid-dose) and 124 mg/kg (high-dose) after H22 tumor cells were implanted. At the end of the experiments, the tumor weight of each mouse was measured. Levels of serum TNF-α and IFN-γ was assayed by ELISA. Protein expressions of Bax, Bcl-2 and caspases-3 were detected by immunohistochemistry. Results Chemical analysis revealed the presence of 6 components that account for 97.55% of fatty acids, indicating the occurrence of saturated and polyunsaturated fatty acids. Oral administration of ESEE could inhibit tumor growth, promote Th1 type cytokine productions (TNF-α and IFN-γ) and induce apoptosis of hepatocarcinoma via increase of Bax/Bcl-2 ratio and activation of caspases-3. Oral administration of ESEE in a dosage of 6.2 g/kg did not lead to toxic effects in mice. Conclusions ESEE was effective in inhibiting tumor growth in vivo and could also serve as immunoadjuvant for tumor therapy.

Sexual size dimorphism (SSD) is a common phenomenon in animals and varies widely among species and among populations within species. Much of this variation is likely due to variance in selection on females vs. males. However, environmental variables could have different effects on females vs. males, causing variation in dimorphism. In this study, we test the differential-plasticity hypothesis, stating that sex-differential plasticity to environmental variables generates among-population variation in the degree of sexual dimorphism. We examined the effect of temperature (22, 25, 28, and 31 °C) on sexual dimorphism in four populations of the cockroach Eupolyphaga sinensis Walker (Blattaria: Polyphagidae), collected at various latitudes. We found that females were larger than males at all temperatures and the degree of this dimorphism was largest at the highest temperature (31 °C) and smallest at the lowest temperature (22 °C). There is variation in the degree of SSD among populations (sex * population interaction), but differences between the sexes in their plastic responses (sex*temperature interaction) were not observed for body size. Our results indicated that sex-differential plasticity to temperature was not the cause of differences among populations in the degree of sexual dimorphism in body size.

Tumor growth and metastasis are responsible for most cancer patients' deaths. Here, we report that eupolyphaga sinensis walker has an essential role in resisting hepatocellular carcinoma growth and metastasis. Compared with proliferation, colony formation, transwell assay and transplantable tumor in nude mouse in vitro and vivo, eupolyphaga sinensis walker extract (ESWE) showed good inhibition on the SMMC-7721 cell growth and metastasis. Using genome-wide microarray analysis, we found the down-regulated growth and metastasis factors, and selected down-regulated genes were confirmed by real-time PCR. Knockdown of a checkpoint PKCβ by siRNA significantly attenuated tumor inhibition and metastasis effects of ESWE. Moreover, our results indicate ESWE inhibits HCC growth by not only downregulating the signaling of PKCβ, Akt, m-TOR, Erk1/2, MEK-2, Raf and JNK-1, but also increasing cyclin D1 protein levels and decreasing amount of cyclin E, cyclin B1 and cdc2 of the cycle proteins. At the same time, ESWE reduced MMP2, MMP9 and CXCR4, PLG, NFκB and P53 activities. Overall, our studies demonstrate that ESWE is a key factor in growth and metastasis signaling inhibitor targeting the PKC, AKT, MAPK signaling and related metastasis signaling, having potential in cancer therapy.