Construction and expression of lentiviral vector of p62 gene over expression / 安徽医科大学学报

Objective @#To construct lentiviral vector of p62 gene over expression , and stably express p62 gene in human monocytic leukemia cells 1 (THP 1) , and to provide a way to study the role of p62 gene at the cellular lev el .@*Methods @#The p62 gene fragment was amplified by polymerase chain reaction (PCR) , and the amplified product was ligated to the linearized pcDNA3 1 Flag PCDH10 lentiviral vector. After identifying with PCR , the PCR product was cotransfected with the packaging plasmid into human embryonic kidney cells 293 (HEK 293T) . THP 1 cells were infected with recombinant lentivirus . Positive cell clones were screened by ampicillin . Western blot and real time fluorescence quantitative polymerase chain reaction (RT qPCR) were used to detect THP 1 cell lines with high p62 expression ( overexpression group) and THP 1 cell lines transfected with empty plasmid without p62 gene ( control group) . The expression levels of TNF α, IL 1βand Cxcl1 after K. p. infection were detected by RT qPCR .@*Results @#The p62 gene fragment was successfully obtained by PCR and ligated to pcDNA3 1 Flag PCDH10 vector. PCR confirmed that p62 pcDNA3 1 Flag PCDH10 recombinant plasmid was constructed successfully. Am picillin resistant cell lines were selected after lentiviral infection of THP 1 cells . The results of Western blot analysis showed that the THP 1 cells with drug sieve survival increased the expression of P62 protein compared with the con trol cells (P < 0.001) , and RT qPCR analysis showed that the relative mRNA expression of p62 increased (P < 0.001) . THP 1 cells with high expression of P62 were successfully constructed . The levels of TNF-α、IL-1βand C xcl1 from THP 1 cells with high expression of P62 significantly increased after infection with K. p. (P < 0.01) . @*Conclusion @#P62 pcDNA3 1 Flag PCDH10 vector and THP 1 cells with high expression of P62 can be successful ly constructed by three plasmid packaging system , which provides a basis for the study of p62 .

Molecular diagnostic strategies and management of dust mite allergy / 中华预防医学杂志

Dust mites are one of the most important allergens, widely distributed around the world, especially in household environments. Dermatophagoides pteronyssinus, Dermatophagoides farinae and Blomia tropicalis are the most common species of dust mites. There are more than 35 known sensitization components of dust mites, among which Der p 1, Der p 2 and Der p 23 are the major components. Clinically, allergen skin test and serum specific immunoglobulin E (sIgE) detection are widely used in the preliminary diagnosis of dust mite allergy. However, these methods cannot accurately identify specific dust mite sensitization components. Considering that there are significant differences in the allergenic components of dust mites in different regions and populations, component-resolved diagnosis of dust mite is particularly important in accurately determining the allergenic components. This is not only of guiding significance for allergen avoidance, but also important for determining the immunotherapy regimen for dust mites. In order to strengthen the understanding of the molecular diagnosis of dust mites and promote the integration of allergy science in China with the international standards, this article interprets the "Allergy Molecular Allergology User's Guide 2.0" published recently by the European Academy of Allergy and Clinical Immunology.

Molecular diagnostic strategies and management of dust mite allergy / 中华预防医学杂志

Dust mites are one of the most important allergens, widely distributed around the world, especially in household environments. Dermatophagoides pteronyssinus, Dermatophagoides farinae and Blomia tropicalis are the most common species of dust mites. There are more than 35 known sensitization components of dust mites, among which Der p 1, Der p 2 and Der p 23 are the major components. Clinically, allergen skin test and serum specific immunoglobulin E (sIgE) detection are widely used in the preliminary diagnosis of dust mite allergy. However, these methods cannot accurately identify specific dust mite sensitization components. Considering that there are significant differences in the allergenic components of dust mites in different regions and populations, component-resolved diagnosis of dust mite is particularly important in accurately determining the allergenic components. This is not only of guiding significance for allergen avoidance, but also important for determining the immunotherapy regimen for dust mites. In order to strengthen the understanding of the molecular diagnosis of dust mites and promote the integration of allergy science in China with the international standards, this article interprets the "Allergy Molecular Allergology User's Guide 2.0" published recently by the European Academy of Allergy and Clinical Immunology.

Analysis of cerebral gray matter structure in multiple sclerosis and neuromyelitis optica / 解剖学报

Objective The volume and cortical thickness of gray matter in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) were compared and analyzed by voxel⁃based morphometry (VBM) and surface⁃based morphometry (SBM), and the differences in the structural changes of gray matter in the two diseases were discussed. Methods A total of 21 MS patients, 16 NMO patients and 19 healthy controls were scanned by routine MRI sequence. The data were processed and analyzed by VBM and SBM method based on the statistical parameter tool SPM12 of Matlab2014a platform and the small tool CAT12 under SPM12. Results Compared with the normal control group (NC), after Gaussian random field (GRF) correction, the gray matter volume in MS group was significantly reduced in left superior occipital, left cuneus, left calcarine, left precuneus, left postcentral, left central paracentral lobule, right cuneus, left middle frontal, left superior frontal and left superior medial frontal (P<0. 05). After family wise error (FWE) correction, the thickness of left paracentral, left superiorfrontal and left precuneus cortex in MS group was significantly reduced (P<0. 05). Compared with the NC group, after GRF correction, the gray matter volume in the left postcentral, left precentral, left inferior parietal, right precentral and right middle frontal in NMO group was significantly increased (P<0. 05). In NMO group, the volume of gray matter in left middle occipital, left superior occipital, left inferior temporal, right middle occipital, left superior frontal orbital, right middle cingulum, left anterior cingulum, right angular and left precuneus were significantly decreased (P<0. 05). Brain regions showed no significant differences in cortical thickness between NMO groups after FWE correction. Compared with the NMO group, after GRF correction, the gray matter volume in the right fusiform and right middle frontal in MS group was increased significantly(P<0. 05). In MS group, the gray matter volume of left thalamus, left pallidum, left precentral, left middle frontal, left middle temporal, right pallidum, left inferior parietal and right superior parietal were significantly decreased (P<0. 05). After FWE correction, the thickness of left inferiorparietal, left superiorparietal, left supramarginal, left paracentral, left superiorfrontal and left precuneus cortex in MS group decreased significantly (P<0. 05). Conclusion The atrophy of brain gray matter structure in MS patients mainly involves the left parietal region, while NMO patients are not sensitive to the change of brain gray matter structure. The significant difference in brain gray matter volume between MS patients and NMO patients is mainly located in the deep cerebral nucleus mass.

Relationship between 24 hour movement behavior and physical fitness among pupils / 中国学校卫生

Objective@#To examine the relationship between 24 hour movement behavior and physical fitness of pupils by using compositional analyses, so as to provide a theoretical basis for physical activity promotion and physical fitness improvement.@*Methods@#By random cluster sampling method, 120 students of grade 4 and 5 from the Second Experimental Primary School of Yingze District of Taiyuan were selected. Physical fitness was evaluated by the indicators of the Protocol of National Physical Training Standard and 20 meter shuttle run. The movement behavior was measured by accelerometer. The relationship between each behavior and physical fitness was analysed by component multiple linear regression, and the effect of replacement of components on physical fitness was discussed.@*Results@#Girls 1minute situps were lower than boys（24.79±7.77，28.21±6.52），and sitting forward flexion was higher than boys[9.00（5.00，14.00），5.20（1.00，9.75）cm]（ t/Z =2.60，-3.15， P ＜0.05）. Boys showed higher light physical activity(LPA), moderate to vigorous physical activity(MVPA) and lower sedentary behavior(SB), while girls showed lower LPA, MVPA and higher SB. MVPA was positively correlated with the performance of 30 second rope skipping and 20 meter shuttle run ( β= 13.19, 7.90, P <0.05). Sleep(SLP), SB, and LPA were not significantly correlated with physical fitness. After re allocating 10 min MVPA to SB and SLP，the performance of 30 second rope skipping and 20 meter shuttle run increased by 2.25，2.28 and 1.28 ,1.34 times，respectively，while significantly decreased after reverse reallocation ( P <0.05). MVPA replaced LPA，the 20 meter shuttle run performance increased 1.46 times，while decreased significantly in reverses ( P <0.05).@*Conclusion@#MVPA is positively correlated with the speed and endurance of pupils. There are gender differences in movement behavior patterns and physical fitness. Children, especially girls, should be encouraged to raise exercise awareness and increase PA, especially MVPA.

Advances in applications of molecular dynamics simulation techniques to the research of self-assembled nano-drug delivery systems / 药学学报

Molecular dynamics simulation technology relies on Newtonian mechanics to simulate the motion of molecular system of the real system by computer simulation. It has been used in the research of self-assembly processes illustration and macroscopic performance prediction of self-assembly nano-drug delivery systems (NDDS) in recent years, which contributes to the facilitation and accurate design of preparations. In this review, the definitions, catalogues, and the modules of molecular dynamics simulation techniques are introduced, and the current status of their applications are summarized in the acquisition and analysis of microscale information, such as particle size, morphology, the formation of microdomains, and molecule distribution of the self-assembly NDDS and the prediction of their macroscale performances, including stability, drug loading capacity, drug release kinetics and transmembrane properties. Moreover, the existing applications of the molecular dynamic simulation technology in the formulation prediction of self-assembled NDDS were also summarized. It is expected that the new strategies will promote the prediction of NDDS formulation and lay a theoretical foundation for an appropriate approach in NDDS studies and a reference for the wider application of molecular dynamics simulation technology in pharmaceutics.

Progress in research of long-term protective efficacy of human papillomavirus vaccine / 中华流行病学杂志

The efficacy of HPV vaccine in preventing cervical cancer has been demonstrated in numerous clinical trials and clinical uses. The follow-up after clinical trials usually last for 5-6 years to evaluate the long-term efficacy, and a series of long-term follow-up studies have been conducted in some regions. The literature retrieval of HPV vaccine long term efficiency research both at home and abroad indicated that the protective efficacy of the vaccine against vaccine-type-related cervical intraepithelial neoplasia grade 2 and above is higher than 90%.

Modern research progress in external application of traditional Chinese medicine to acupoints / 中国中药杂志

Traditional Chinese medicine(TCM) has a long history and abundant experience in external therapy, which marks human wisdom. In the early history of human, people found that fumigation, coating, and sticking of some tree branches and herb stems can help alleviate scabies and remove parasites in productive labor, which indicates the emergence of external therapy. Pathogen usually enters the body through the surface, so external therapy can be used to treat the disease. External therapy is among the major characteristic of surgery of TCM. As one of the external therapies in TCM, external application to acupoints smooths the zang-fu organs through meridians and collaterals, thereby harmonizing yin and yang. This therapy emerged in the early society, formed the Spring and Autumn Period and the Warring States Period, improved in the Song and Ming dynasties, and matured in the Qing dynasty. With the efforts of experts in history, it has had a mature theory. According to modern research, it can avoid the first-pass effect of liver and the gastrointestinal irritation and improve the bioavailability of Chinese medicine. Based on the effect of Chinese medicine and the theory of meridian and collateral, it can stimulate the acupoints, exert regulatory effect on acupoints, and give full play to the efficacy of TCM and the interaction of the two. Thereby, it can regulate qi and blood and balance yin and yang, thus being widely used in the treatment of diseases. In this paper, the use of external application to acupoints, the effect on skin immunity, the regulation of neuro-inflammatory mechanism, the relationship between acupoint application and human circulation network, and the development of its dosage form were summarized through literature review. On this basis, this study is expected to lay a foundation for further research.

Effect of apigenin in combination with oxymatrine on non-small cell lung cancer and mechanism / 中国中药杂志

This study explores the effect of apigenin(APG), oxymatrine(OMT), and APG+OMT on the proliferation of non-small cell lung cancer cell lines and the underlying mechanisms. Cell counting kit-8(CCK-8) assay was used to detect the vitality of A549 and NCI-H1975 cells, and colony formation assay to evaluate the colony formation ability of the cells. EdU assay was employed to examine the proliferation of NCI-H1975 cells. RT-qPCR and Western blot were performed to detect the mRNA and protein expression of PLOD2. Molecular docking was carried out to explore the direct action ability and action sites between APG/OMT and PLOD2/EGFR. Western blot was used to study the expression of related proteins in EGFR pathway. The viability of A549 and NCI-H1975 cells was inhibited by APG and APG+OMT at 20, 40, and 80 μmol·L~(-1) in a dose-dependent manner. The colony formation ability of NCI-H1975 cells was significantly suppressed by APG and APG+OMT. The mRNA and protein expression of PLOD2 was significantly inhibited by APG and APG+OMT. In addition, APG and OMT had strong binding activity with PLOD2 and EGFR. In APG and APG+OMT groups, the expression of EGFR and proteins in its downstream signaling pathways was significantly down-regulated. It is concluded that APG in combination with OMT could inhibit non-small lung cancer, and the mechanism may be related to EGFR and its downstream signaling pathways. This study lays a new theoretical basis for the clinical treatment of non-small cell lung cancer with APG in combination with OMT and provides a reference for further research on the anti-tumor mechanism of APG in combination with OMT.

Effects of shading intensity on growth and quality of Artemisia stolonifera / 中国中药杂志

The purpose of this study was to analyze the effects of shading intensity on the growth, yield, and quality of Artemisia stolonifera so as to provide references for the artificial cultivation of A. stolonifera. The seedlings of A. stolonifera with consistent growth underwent shading treatment at four shading intensity levels(0, 55%, 85%, and 95%) with different layers of black shading nets. The agronomic indexes, yield, moxa yield, total ash, quality characteristics of moxa during combustion and pyrolysis, main volatile components, flavonoids, and phenolic acids were measured. The results showed that under shading conditions, the stem diameter, leaf width, 5-leaf spacing, branch number, and yield of A. stolonifera decreased significantly, while the plant height, leaf length, leaf number, chlorophyll content, and moxa yield increased first and then decreased with the increase in shading intensity. The burning performance of moxa under natural light was better than that under moderate and severe shading conditions. The content of eucalyptol first increased and then decreased with the increase in shading intensity. The humulene content was negatively correlated with shading intensity. Other major volatile components showed no significant difference under various shading conditions. The content of neochlorogenic acid, cryptochlorogenic acid, isoschaftoside, and isochlorogenic acid B was positively correlated with shading intensity, while the content of chlorogenic acid, isochlorogenic acid A, and isochlorogenic acid C decreased first and then increased with the increase in shading intensity. To sum up, A. stolonifera is a light-loving plant, and shading can greatly reduce the yield, the content of internal components, and the burning performance of moxa. It is the main reason why A. stolonifera is mainly distributed in the forest edge, open forest, roadside, and wasteland grass in the middle and high mountains in the wild. For artificial domestication and cultivation of A. stolonifera, it is better to select plots with sufficient light.

Comparison of growth and quality of wild and cultivated Artemisia stolonifera / 中国中药杂志

This paper aims to compare the difference of growth and quality between wild and cultivated Artemisia stolonifera, thereby providing references for further development and utilization of A. stolonifera. The wild and cultivated A. stolonifera from different altitudes were collected, and the agronomic characters, moxa yield, volatile components, flavonoids, and phenolic acids were determined. The results showed that the cultivated species were taller and stronger, with more leaves and branches, than the wild species. The moxa yield and combustion quality of wild products were higher than those of cultivated products. The content of main volatile components in cultivated products was higher than that in wild products. The content of flavonoids and phenolic acids in wild products was higher than that in cultivated products. At high altitude, the ignition performance, combustion persistence, comprehensive combustion performance, and heat release during combustion of the wild and cultivated A. stolonifera. were optimal. At middle altitude, the content of main characteristic volatile components and flavone phenolic acids in the leaves of the cultivated and wild A. stolonifera were the highest. At low altitude, the combustion quality and the content of the above components of the cultivated A. stolonifera decrease significantly. Considering the combustion quality and the content of the internal components of the leaf lint, the middle and high altitude areas are suitable for the artificial cultivation of A. stolonifera.

Application research and design strategy on smart responsive mesoporous silica anti-tumor nanodelivery systems / 药学学报

Malignant tumors are major diseases that endanger human health. Due to their complex and variable microenvironment, most anti-tumor drugs cannot precisely reach the focal tissue and be released in a controlled manner. Intelligent responsive nano carriers have become a hot spot in the field of anti-tumor drug delivery systems. As an excellent nano material, mesoporous silica has the advantages of non-toxic, stable, adjustable pore volume and pore diameter, and easy functional modification on the surface. By virtue of its perceptive response to the tumor microenvironment or physiological changes, it can achieve the targeted drug release or controlled drug release of the drug delivery system in the tissue, making it an ideal carrier for intelligent response drug delivery system. In this paper, we review the design strategies and current research status of smart responsive anti-tumor drug delivery systems based on mesoporous silica, in order to provide a reference for the development of anti-tumor drug nanoformulations.

Comparison of a minimally invasive osteosynthesis technique with conventional open surgery for transverse patellar fractures / 中华创伤杂志(英文版)

PURPOSE@#The study aims to compare the efficacy and safety of a new minimally invasive osteosynthesis technique with those of conventional open surgery for transverse patellar fractures.@*METHODS@#It was a retrospective study. Adult patients with closed transverse patellar fracture were included, and with open comminuted patellar fracture were excluded. These patients were divided into minimally invasive osteosynthesis technique (MIOT) group and open reduction and internal fixation (ORIF) group. Surgical time, frequency of intraoperative fluoroscopy, visual analogue scale score, flexion, extension, Lysholm knee score, infection, malreduction, implant migration and implant irritation in two groups were recorded and compared. Statistical analysis was performed by the SPSS software package (version 19). A p < 0.05 indicated statistical significance.@*RESULTS@#A total of 55 patients with transverse patellar fractures enrolled in this study, the minimally invasive technique was performed in 27 cases, and open reduction was performed in 28 cases. The surgical time in the ORIF group was shorter than that in the MIOT group (p = 0.033). The visual analogue scale scores in the MIOT group were significantly lower than those in the ORIF group only in the first month after surgery (p = 0.015). Flexion was restored faster in the MIOT group than that in the ORIF group at one month (p = 0.001) and three months (p = 0.015). Extension was recovered faster in the MIOT group than that in the ORIF group at one month (p = 0.031) and three months (p = 0.023). The recorded Lysholm knee scores in the MIOT group were always greater than those in the ORIF group. Complications, such as infection, malreduction, implant migration, and implant irritation, occurred more frequently in the ORIF group.@*CONCLUSION@#Compared with the ORIF group, the MIOT group reduced postoperative pain and had less complications and better exercise rehabilitation. Although it requires a long operation time, MIOT may be a wise choice for transverse patellar fractures.

Analysis of AHNAK2 gene polymorphism in Xinjiang patients with coronary heart disease / 心血管康复医学杂志

Objective:To explore the relationship between the polymorphisms of AHNAK2 gene rs12882641,rs28583515 and rs2582497 loci and coronary heart disease(CHD)in the population of Xinjiang.Materials:This study used a case-control method,a total of 602 patients who were hospitalized and underwent coronary angiogra-phy(CAG)at ourheart center from Jan 2019 to Dec 2021 were selected.According toCAG results,the patients were divided into CHD group(n=301)and non-CHD group(n=301).The AHNAK2 gene rs12882641,rs28583515 and rs2582497 loci were genotyped using the improved multiple ligase detection reaction(iMLDR)technique,and the relationship between AHNAK2 gene polymorphisms and CHD was analyzed.Results:Compared with non-CHD group,there was significant rise in the distribution frequencies of AC+CC genotypes(52.8%vs.61.1%,P= 0.040)at rs2582497 locus of the AHNAK2 gene under the dominant model;there was significant reduction in distri-bution frequency of the CC genotype(65.8%vs.53.8%)at the rs28583515 locus of the AHNAK2 gene,and signif-icant rise in distribution frequencies of CT+TT(34.2%vs.46.2%)under the dominant model,TT under the re-cessive model(0.7%vs.3.0%)and CT under the additive model(33.6%vs.43.2%)in CHD group,P＜0.05 or＜0.01.After adjusting for confounding factors,logistic regression analysis indicated that the dominant model of the rs28583515 locus remained an independent risk factor for CHD(OR=1.509,P=0.036).Conclusion:The AH-NAK2 gene rs28583515 locus is closely related to the occurrence and development of CHD in the Xinjiang popula-tion.The dominant model of the AHNAK2 gene rs28583515 locus is an independent risk factor for CHD.

Associations between m 6A RNA methylation and cutaneous melanoma / 中华皮肤科杂志

N 6-methyladenosine (m 6A) modification can regulate gene expression by influencing RNA stability, nuclear transport, translation efficiency, alternative splicing and degradation. Current studies have shown that m 6A modification is associated with multiple biological behaviors of melanoma, such as proliferation, metastasis and resistance to immunotherapy, but specific mechanisms still need further study. This review summarizes the regulation of m 6A RNA methylation, and its related biological behaviors and mechanisms of action in melanoma.

Comparison of immune responses and protective efficacy between mucosal vaccines and intramuscular vaccines / 中华微生物学和免疫学杂志

The routes of immunization largely determine the features of vaccine-induced immune responses. This paper mainly reviewed the immune responses and protective efficacy induced by mucosal immunization and intramuscular injection of vaccines against poliovirus, Salmonella typhi, influenza virus and SARS-CoV-2 and compared the differences between different immunization routes, with the goal of providing some fundamental theoretical data for the development of vaccines and vaccination strategies against infectious diseases.

Effectiveness of HPV vaccines in real-world study / 中华微生物学和免疫学杂志

Human papillomavirus (HPV) is an epitheliotropic virus. High-risk HPV infections lead to precancerous lesions which may progress to cancer in the cervix, vagina and vulva, while low-risk HPV infections cause benign lesions such as genital warts and recurrent respiratory papillomas. HPV infection remains one of the major public health problems threatening human health. To date, six prophylactic preventive HPV vaccines have been licensed, and the effectiveness of HPV vaccination has gradually appeared in some countries with earlier vaccination. HPV vaccination has been proved to be effective in protecting against diseases related to HPV infection, which leads to significant reductions in the incidence of vaccine-type HPV-related infection, high cervical lesions, anogenital warts, recurrent respiratory papillomatosis and other relevant diseases. The herd protection effect of the vaccines is outstanding. Meanwhile, a bivalent HPV vaccine has been demonstrated for the cross-protection against HPV infections of non-vaccine types (HPV31/33/45) in real-world vaccination practice.

Research progress in the protective efficacy of human papillomavirus vaccines in males / 中华微生物学和免疫学杂志

Human papillomavirus (HPV) is one of the most common sexually transmitted pathogens. It can cause a variety of diseases such as condyloma acuminatum, anal cancer, penile cancer and oropharyngeal cancer in men, resulting in a high disease burden. With the development of society, the application of HPV vaccines in males has attracted more attention. Currently, there are many clinical trials and real-world research results of HPV vaccines applied to boys and men worldwide, and many countries have introduced HPV vaccination for underage boys into their national immunization programs. This article intended to review the research progress in the efficacy of HPV vaccines in male population.

Effect and molecular mechanism of mammalian target of rapamycin complex 2 / Akt signaling pathway on 6-hydroxydopamine-treated SH-SY5Y cell model / 解剖学报

[Abstract] Objective To study whether the regulation of mammalian target of rapamycin complex 2(mTORC2) / Akt signaling pathway has a protective effect on SH-SY5Y cell line damaged by 6-hydroxydopamine (6-OHDA), and to clarify its molecular mechanism. Methods SH-SY5Y cells treated with retinoic acid (RA) were given 6-OHDA, mTORC2 signaling pathway inhibitor PP242 and agonist A-443654 respectively. The changes of cell number in each group were investigated by immunofluorescent staining; The total protein was extracted and the expression level and interaction of key proteins in mTORC2 signaling pathway were determined by Western blotting and co-immunoprecipitation (CoIP); The apoptosis rate of cells in each group was detected by flow cytometry. At the same time, the co-culture Parkinson’ s disease (PD) model was made using SH-SY5Y cell line and Bv-2 cell line; MTT colorimetric method was used to detect the cell viability of each group; ELISA was used to detect the content of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in cell culture supernatant. Results The number of tyrosine hydroxylase(TH) / proliferating cell nuclear antigen (PCNA) / hochest-, TH / 5-bronmo-2’ -deoxyuridine(BrdU) -labeled positive cells in 6-OHDA-lesioned PD cell model group was significantly lower than that in the normal group; The apoptosis rate was higher; The expression of Rictor, p-Akt and regulated in DNA damage and development 1(REDD1) was increased; There was an interaction between Rictor and p-Akt or REDD1; The cell viability was significantly reduced in the co-culture model; the content of TNF-α and IL-β increased in the cell culture supernatant. With further up-regulation of the abovementioned protein expressions, the cell survival, apoptosis and pro-inflammatory cytokine levels in A-443654 group were significantly ameliorated, while PP242 group showed the opposite changes. Conclusion A-443654 activates mTORC2 signaling pathway by p-Akt, which increases the expression of Rictor and REDD1 protein. These changes contribute to the amelioration in cell survival rate, apoptosis rate, and the proliferation and differentiation and decreasion of apoptosis rate of SH-SY5Y cells. These result improved 6-OHDA-induced cell damage and inhibited the release of pro-inflammatory cytokines.

Effects of activating mTORC2/Akt signaling pathway on dopaminergic neurons and behaviors in 6-hydroxydopamin model mice / 解剖学报

Objective To explore the effect of activation of mammalian target of rapmycin complex 2(mTORC2)/Akt signaling pathway on dopaminergic neurons and behavior in 6-hydroxydopamine (6-OHDA) model mice and its possible mechanism. Methods Selecting 36 mice which The Nestin-CreERTM and ROSA26-LacZ reporter genes were detected at the same time in 3-month-old male C57BL/6J mice weighing 20-25 g divideng them into 4 gruops, NS+ corn oil group, 6-OHDA+corn oil group, 6-OHDA+PP242 group and 6-OHDA+A-443654 group, and 6-OHDA was injected into the right striatum of the brain to replicate the Parkinson’s disease (PD) model of mice, and then daily intraperitoneal injection of mTORC2/Akt signaling pathway agonist A-443654 or inhibitor PP242. Serum interleukin-1β (IL-1β) and tumor necrosis factor-α(TNF-α)levels were measured by enzyme-linked immunosorbent assay. Immunohistochemistry and immunofluorescence staining were performed to investigate the change of microglia, dopaminergic neurons as well as neural progenitor cells (NPCs). Western blotting was used to detect the expression of related protein of mTORC2/Akt signaling pathway including rictor, p-Akt and regulated in development and DNA dgmage responses 1(REDD1) and the interaction between them were verified by immunoprecipitation. Finally, the behavioral performance of each group of mice was observed. Results With the activation of microglia and the increase of inflammatory factors in PD model mice, the number of dopaminergic neurons in the substantia nigra(SN) decreased significantly, and the motor function of the mice was impaired, but the number of NPCs increased significantly compared with the control mice, mTORC2/Akt signaling pathway related protein expression was also significantly up-regulated. A-443654 treatment further up-regulated the expression of these proteins, meanwhile the indicators mentioned above were ameliorated. However, the inhibitor PP242 treatment group showed completely opposite result with the agonist group. Conclusion A-443654 can promote the proliferation of NPCs and the number of new-born dopaminergic neurons by up-regulating related proteins of mTORC2/Akt signaling pathway, and reducing the activation of microglia and the level of inflammation factors, which ultimately lead to the amelioration of SN-striatal dopaminergic neurons and behavioral performance in PD model mice.