Involvement of PI 3 K/Akt/mTOR Signaling in Protective Effects of Moxibustion for Premature Ovarian Failure in Rats / 针刺研究

OBJECTIVE: To study the protective effect of moxibustion for tripterygium-induced premature ovarian failure (POF) and its underlying mechanisms in rats. METHODS: Forty-five female SD rats were randomly divided into normal control, POF model and moxibustion groups (n＝15/group). The POF model was induced by intragastric administration of Triptolide (40 mg/kg), once daily for 6 weeks. From the 4th week after modeling, moxibustion was given at "Guanyuan" (CV 4) and bilateral "Sanyinjiao" (SP 6) for 10 min, once daily for 3 weeks. Pathological changes of ovary tissues were determined by hematoxylin-eosin (HE) staining. The serum estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), interleukin-6 (IL-6) and interleukin-1 β (IL-1 β) contents were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of phosphatidyl inositol 3- kinase (PI 3 K), protein kinase B (Akt) and mammalian target of rapamycin (mTOR) proteins of the ovarian tissue were detected by Western blot. RESULTS: After modeling, HE staining showed that the numbers of ovarian follicles and follicular granulocytes and corpora luteum layers were decreased, and the number of corpora atretica was increased in the model group. The content of serum E2 was markedly decreased and those of serum LH, FSH, IL-6 and IL-1 β were markedly increased in the model group (P<0.01), and the expression levels of ovarian p-PI 3 K, p-Akt and p-mTOR were markedly increased after modeling relevant to the control group (P<0.01). Following moxibustion, the pathological damage of ovarian tissue was improved, the contents of serum LH, FSH, IL-6, IL-1 β, and the levels of p-PI 3 K, p-Akt and p-mTOR proteins in the ovarian tissue were significantly decreased (P<0.05, P<0.01), and the content of serum E2 was markedly increased (P<0.05) in comparison with the model group. CONCLUSION: Moxibustion can improve POF in POF rats, which may be related to its actions in inhibiting PI 3 K/Akt/mTOR signaling, down-regulating serum IL-6, IL-1 β, and regulating serum hormones.

Mechanism of Xuebijing Injection in anti-acute lung injury based on network pharmacology / 中草药

Objective: To predict the action targets of anti-acute lung injury active ingredients of Xuebijing Injection, and investigate the “multi-components, multi-targets, and multi-pathways” mechanism. Methods: Reverse molecular docking was used to forecast its targets of 38 main active components of Xuebijing Injection. The relevant targets of acute lung injury were searched through literature mining and multiple databases and compared with the predicted component targets. The protein interaction network was constructed by Cytoscape software, and topological analysis was performed to screen the key targets of anti-acute lung injury of Xuebijing Injection. And GO enrichment analysis and KEGG analysis were carried out on the key targets through DAVID database. Results: The network analysis indicated that 38 active ingredients affected 143 key target proteins directly or indirectly, involved in 71 biological processes, 29 cellular components, 40 molecular function, and 25 KEGG pathways. These active compounds might participate in regulating the processes of gene, protein, external stimulus, and interfering the mitogen-activated protein kinase (MAPK), phosphatidyl inositol 3-kinase (PI3K)-Akt, and other signaling pathways to play a role in resisting to acute lung injury. Conclusion: The anti-acute lung injury effect of Xuebijing Injection showed the characteristics of traditional Chinese medicine in multi-components, multi-targets, and multi-pathways. This research provides a scientific basis for elucidation of the anti-acute lung injury pharmacological mechanism of Xuebijing Injection.

Effect of Chemotatic Factor FKN,PI3K and NF-κB on IL-6 Expression in Peripheral Blood Monocytes and the Effect of Valsartan Intervention / 中国医科大学学报

Objective To explore effect of irregular chemotatic factor fractalkine(FKN),phosphatidyl inositol 3 kinase and nuclear factor?κB(NF?κB)on interleukin?6(IL?6)expression in peripheral blood monocytes and the effect of valsartan intervention,so as to research the signal conduc?tive mechanism of FKN impacting on IL?6. Methods Peripheral blood monocytes were isolated from fresh blood of healthy volunteers by the densi?ty gradient centrifugation. The extractive peripheral blood monocytes were divided into seven groups:the control group,the FKN group,the LY294002 group(PI3K inhibitors),the PDTC group(NF?κB inhibitors),the FKN+valsartan group,the FKN+LY294002 group,and the FKN+PDTC group,the latter two were pretreated by LY294002 and PDTC respectively before FKN inducing PBMC cells. The IL?6 expression in cell me?dium was measured in each group by ELISA at 12 hours and 24 hours after PBMC treatment. Results After 12 hours of culture,compared with the control group,the expression of IL?6 in the FKN group was decreased(P0.05). Compared with the FKN group,the expression of IL?6 was decreased in the FKN+valsartan group(P0.05). Conclusion FKN can adjust the expression of peripheral blood PBMC IL?6 in a two?way pattern, inhibiting the expression of IL?6 by PI3K pathway and promoting the expression of IL?6 by NF?κB pathway,overall,FKN can inhibit the expression of IL?6. Valsartan can increase FKN to inhibit the expression of IL?6.

Effect of QizhiJiangtang Capsule on insulin resistance in diabetic rats and its mechanism / 吉林大学学报(医学版)

Objective To explore the effect of QizhiJiangtang Capsule on the insulin resistance (IR)in the diabetic rats,and to clarify the action mechanism.Methods The diabetes rat models were induced by high fat diet combined with STZ injection.The successful models of the rats were randomly divided into diabetes group (DM), ShenqiJiangtang Granule group (SQ)and high (QJH),middle-(QJM),low (QJL)doses of QizhiJiangtang Capsule groups;at the same time control group (NC)was established. The drug concentrations in high, middle and low-doses of QizhiJiangtang Capsules groups were 1.35, 0.68, and 0.34 g · kg-1 respectively;and the concentration of ShenqiJiangtang Granule was 0.27 g·kg-1.After the diabetic model was established successfully, the rats were treated for 8 weeks on the basis of drug dose.Then the levels of fasting blood glucose (FBG),fasting insulin (FINS),insulin resistance index (IRI)and biochemical indexes related to lipid metabolism of the rats were measured using blood glucose detector and automatic biochemistry analyser.The gene expression of insulin receptor substrate-1 (IRS-1),phosphatidyl inositol 3-kinase (PI3K),and glucose transporter 4 (GLUT4)in liver tissue were examined by Real Time PCR.The levels of tumor necrosis factorα(TNF-α)and adiponectin (ADPN)in serum were detected using ELISA.Results Compared with control group,the levels of FBG,FINS and IRI of the rats in diabetes group were significantly increased (P<0.05 or P<0.01 );the serum total cholesterol (TC), triglyceride (TG)and low density lipoprotein (LDL)levels were significantly increased (P<0.05 ), while the serum high-density lipoprotein (HDL)level was significantly decreased (P<0.05);the mRNA expression levels of IRS-1,PI3K and GLUT4 in liver tissue were decreased (P<0.05);the level of serum TNF-αwas increased (P<0.05),but the ADPN level was decreased (P<0.05).Compared with diabetes group,the FBG level and IRI of the rats in QizhiJiangtang Capsule and ShenqiJiangtang Granule groups were significantly decreased (P<0.01);the levels of FINS of the rats middle and high doses of in QizhiJiangtang Capsule groups and ShenqiJiangtang Granule group were significantly decreased (P<0.05);the levels of serum TC,TG and LDL of the rats in middle dose of QizhiJiangtang Capsule group and ShenqiJiangtang Granule group were significantly decreased (P<0.05 or P<0.01),but the HDL level was increased (P<0.05);the mRBA expression lvels of IRS-1,PI3K and GLUT4 inliver tissue were increased (P<0.05);the levels of serum TNF-αof the rats in middle dose of QizhiJiangtang Capsule group and Shenqijiangtang Granule group were significantly decreased (P<0.05),but the serum ADPN levels were increased (P<0.05 ). Conclusion QizhiJiangtang Capsule can significantly improve the IR in the diabetic rats,and the pharmacological mechanisms are related to adapting the blood lipid component and insulin signal transduction pathways.

Profilin activates bacillus thuringiensis phosphoinositide specific phospholipase C.

Many extracellular signaling molecules including hormones, growth factors, neurotransmitters and immunoglobulins elicit intracellular responses by activating phosphatidylinositol-specific phospholipase C (PI-PLC) upon binding to their cell surface receptors. Activated PLC catalyses the hydrolysis of Phosphotidylinositol 4,5- bisphosphate (PIP2) to generate DAG and IP3 , which act as signaling molecules that control various cellular processes. Exploring the mechanism of regulation of PLC activity may lead to understanding various signaling events that regulate cell growth and differentiation. One of the dramatic effects of profilin is inhibition of PIP2 hydrolysis by PLC- γ in eukaryotic cells. In the present study, the effect of profilin on Phosphotidylinositol specific phospholipase C (PIPLC) purified from Bacillus thuringiensis (Bt) was examined. Assay of PI-PLC activity indicated that Bovine profilin activated the hydrolysis of phosphotidylinositol (PI) by BtPI-PLC in a concentration dependent manner under in vitro conditions. A 250 % increase in activity was noted in the presence of profilin but not in presence of phosphoprofilin. In the presence of profilin more proteins are observed in the soluble fraction. In conclusion it can be stated that that profilin activates bacterial PLC activity towards PI hydrolysis.

Mutation Analysis of PIG-A Gene in Korean Patients with Paroxysmal Nocturnal Hemoglobinuria / 대한혈액학회지

BACKGROUND: Paroxysmal nocturnal hemoglobinuria (PNH) is caused by deficient biosynthesis of the glycosylphosphatidylinositol (GPI) anchor in hemopoietic stem cells. Mutation of phosphatidyl inositol glycan class A (PIG-A) gene, an X-linked gene that participates in the first step of GPI anchor biosynthesis, is responsible for PNH. Characteristics of somatic mutation of PIG-A gene in the Korean patients with PNH and their relationships to clinical characteristics were analyzed. METHODS: Twenty five patients with PNH and a donor of bone marrow transplantation were selected. Ham tests, sucrose hemolysis tests and CD59 expressions of erythrocytes and granulocytes were performed to confirm diagnosis. Dideoxy fingerprinting (ddF) was used to screen mutations, and direct sequencing of DNA was performed to characterize the mutations. RESULTS: The mutations of PIG-A gene were found in twelve cases and ten of them were novel mutations. There were five deletions, six substitutions and a insertion. Therewere six premature terminations, three abnormal splicings, a missense and two nonsense mutations. There were six point mutations and six frameshift mutations. Five cases of hypoplastic PNH showed mutations only in exons, but three in seven cases of hemolytic PNH showed mutations in introns. Two cases with symptoms of venous thrombosis showed mutations in exon 3. CONCLUSION: There were ten novel mutations among twelve mutations in the Korean patients with PNH and characteristics of the mutations were variable without any remarkable hot spot in sites and types. The characteristics of mutation were not correlated with the results of clinical courses of the patients with PNH.

Study on Clinical and Laboratory Features of Childhood Paroxysmal Nocturnal Hemoglobinuria / 实用儿科临床杂志

Objective To learn more about the clinical and laboratory features of childhood paroxysmal nocturnal hemoglobinuria(PNH) and to improve the diagnosis.Methods The clinical and laboratory features of 12 cases of PNH were analyzed,who were diagnosed from January 2000 to November 2004,and the positive responses to treatments were observed.Results 1) The youngest age of onset was 2 years;the disease often manifested with anemia(100%),recurrent infections(50%) and hemorrhages(33%),occurring mainly in skin and mucosa.No patient developed a thrombosis.2) 66.7% of the patients showed peripheral blood cytopenia.Dysplasia of bone marrow was observed in 25% of patients.Fifty percent of them had an increased percentage of erythroid lineage.3) Positive hemolytic tests included urine OB 41.6%,Ham′s test 33.3% and Rous′ test 25%.Glycosyl-phosphatidyl inositol(GPI) deficient cells were found in 100% of the patients.4) 57.1% of patients was improved after being treated with adrenocortical hormone,androgens or cysporin.Conclusions Besides hemoglobinuria,peripheral blood cytopenia were also the common manifestation of PNH.Flow cytometry based immunophenotypic methods for the analysis of CD_(55) and CD_(59) may improve the diagnosis of PNH.J Appl Clin Pediatr,2006,21(3):153-154