1.Effect of 17-β-estradiol on the expression of glucose transporter 4 in rat primary culture skeletal muscle cells with insulin resistance induced by palmitinic acid
Feike YANG ; Jingfang LIU ; Xiaofang LI ; Jinsha FAN ; Jing ZHOU ; Wei CHEN
Chinese Journal of Geriatrics 2012;(12):1112-1116
Objective To investigate the effect of 17-β-estradiol (17-β-E2) on the expression of glucose transporter 4 (GLUT4) in rat primary culture skeletal muscle cells with insulin resistance (IR) induced by palmitinic acid (PA).Methods Rat skeletal muscle was primarily cultivated.The cells were identified by the technology of immunofluorescence.Cells were inclubated with 0.6 mmol/L palmitinic acid for 24 h to induce IR.The concentration of glucose in the medium was measured.The mRNA and protein expressions of protein kinase B (PKB/Akt) and GLUT4 were measured by real time polymerase chain reaction (PCR) and Western blot respectively.Results Compared the control with PA treated rats,the concentration of glucose in the medium was increased [(3.86±0.64)mmol/L vs.(5.43±0.55) mmol/L,q=4.13,P<0.05],uptake of glucose stimulated by insulin was decreased (P<0.01),the expressions of GLUT4 and phosphorylation PKB/Akt were reduced in the PA group (all P<0.05).In the 17-β-E2 (100 nmol/L) versus control,the concentration of glucose in the medium was higher [(3.86±0.64) vs.(3.77±0.35)mmol/L,q= 4.76,P<0.05],uptake of glucose stimulated by insulin was increased (P<0.01) and the expressions of GLUT4 and p-Akt were also increased (all P<0.05).17-β-E2 (10 nmol/L and 100 nmol/L) reversed the decrease of basal and insulin-stimulated uptake of glucose induced by PA,reversed the decrease of GLUT4 and p-Akt expression induced by PA compared with the PA group (all P< 0.05).Conclusions 17-β-E2 inhibits insulin resistance induced by PA in rat primary culture skeletal muscle cells,the mechanism may be correlated with the up-regulation of expression of phospho-Akt and GLUT 4 induced by 17-β-E2.
2.Mechanism of Dahuang Zhechongwan in Treating Myocardial Fibrosis of Rats via TGF-β1/Smads/miR29 Pathway
Jingtao LIANG ; Xiaoyan HE ; Min WANG ; Zhiying HUO ; Fan GUO ; Lijuan WU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(14):21-29
ObjectiveTraditional Chinese medicine, namely Dahuang Zhechongwan (DHZCW) was used to treat myocardial fibrosis in model rats, observe its effect on myocardial fibrosis in rats, and explore its action mechanism. MethodThirty-six SPF male Kunming rats were divided into blank group, model group, low-, medium-, high-dose groups of DHZCW (0.056, 0.084, 0.168 g·kg-1), captopril group (10 mg·kg-1), with six rats in each group. Except for the blank group, the other groups were intraperitoneally injected isoproterenol solution of 5 mg·kg-1 for 15 consecutive days to replicate the myocardial fibrosis model. At the beginning of modeling, the rats in each group took drugs, and they were sacrificed 28 days after administration. Serum and heart tissue were collected for the corresponding detection. Hematoxylin-eosin (HE) staining and Masson staining were used to observe tissue inflammation, cellular degeneration, necrosis, and fibrosis. The contents of hydroxyproline (HYP), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), hyaluronic acid (HA), laminin (LN), type-Ⅲ procollagen (PC Ⅲ) in serum of rats and rats were determined by enzyme-related immunosorbent assay (ELISA). The expression levels of key pathway proteins transforming growth factor-β1 (TGF-β1), α-smooth muscle actin (α-SMA), Smad2, Smad3, and Smad7 were detected by Western blot. The expression levels of key pathway genes TGF-β1, α-SMA, Smad2, Smad3, Smad7, miR-29a-5p, miR-29b-2-5p, and miR-29c-5p were detected by Real-time quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the blank group, the pathological changes of fibrosis in the model group were obvious, the contents of serum HYP, TNF-α, IL-1β, IL-6, HA, LN, and PCⅢ were increased (P<0.01), the protein expression levels of TGF-β1, α-SMA, Smad2, and Smad3 were increased; the protein expression level of Smad7 was decreased (P<0.01). The mRNA expression levels of TGF-β1, α-SMA, Smad2, and Smad3 were increased (P<0.05, P<0.01), while those of Smad7, miR-29a-5p, miR-29b-2-5p, and miR-29c-5p were decreased (P<0.01). Compared with the model group, after 28 days of administration, serum HYP, TNF-α, IL-1β, IL-6, HA, LN, and PCⅢ in high-, medium-, and low-dose groups of DHZCW and captopril groups were decreased (P<0.01). Except for the low-dose group, the protein contents of TGF-β1, α-SMA, Smad2, and Smad3 were decreased, while the protein content of Smad7 was increased (P<0.01). The mRNA expression levels of TGF-β1, Smad2, α-SMA, and Smad3 in high-dose group of DHZCW were decreased (P<0.05,P<0.01), while those of Smad7, miR-29a-5p, miR-29b-2-5p, and miR-29c-5p were increased (P<0.05). The mRNA expressions of TGF-β1, Smad2, and Smad3 in the medium-dose group of DHZCW were decreased (P<0.05, P<0.01), while mRNA expression of Smad7 was increased (P<0.01). The mRNA levels of TGF-β1 and Smad2 in the low-dose group of DHZCW were decreased (P<0.01). ConclusionDHZCW can improve myocardial fibrosis in rats, and its action mechanism may be related to the regulation of the TGF-β1/Smads/miR-29 pathway. In addition, there is dose dependence in the range of 0.056-0.168 g·kg-1, and the effect of the high-dose group is more stable.
3.Genomic and transcriptomic analysis unveils population evolution and development of pesticide resistance in fall armyworm Spodoptera frugiperda.
Furong GUI ; Tianming LAN ; Yue ZHAO ; Wei GUO ; Yang DONG ; Dongming FANG ; Huan LIU ; Haimeng LI ; Hongli WANG ; Ruoshi HAO ; Xiaofang CHENG ; Yahong LI ; Pengcheng YANG ; Sunil Kumar SAHU ; Yaping CHEN ; Le CHENG ; Shuqi HE ; Ping LIU ; Guangyi FAN ; Haorong LU ; Guohai HU ; Wei DONG ; Bin CHEN ; Yuan JIANG ; Yongwei ZHANG ; Hanhong XU ; Fei LIN ; Bernard SLIPPERS ; Alisa POSTMA ; Matthew JACKSON ; Birhan Addisie ABATE ; Kassahun TESFAYE ; Aschalew Lemma DEMIE ; Meseret Destaw BAYELEYGNE ; Dawit Tesfaye DEGEFU ; Feng CHEN ; Paul K KURIA ; Zachary M KINYUA ; Tong-Xian LIU ; Huanming YANG ; Fangneng HUANG ; Xin LIU ; Jun SHENG ; Le KANG
Protein & Cell 2022;13(7):513-531
The fall armyworm (FAW), Spodoptera frugiperda, is a destructive pest native to America and has recently become an invasive insect pest in China. Because of its rapid spread and great risks in China, understanding of FAW genetic background and pesticide resistance is urgent and essential to develop effective management strategies. Here, we assembled a chromosome-level genome of a male FAW (SFynMstLFR) and compared re-sequencing results of the populations from America, Africa, and China. Strain identification of 163 individuals collected from America, Africa and China showed that both C and R strains were found in the American populations, while only C strain was found in the Chinese and African populations. Moreover, population genomics analysis showed that populations from Africa and China have close relationship with significantly genetic differentiation from American populations. Taken together, FAWs invaded into China were most likely originated from Africa. Comparative genomics analysis displayed that the cytochrome p450 gene family is extremely expanded to 425 members in FAW, of which 283 genes are specific to FAW. Treatments of Chinese populations with twenty-three pesticides showed the variant patterns of transcriptome profiles, and several detoxification genes such as AOX, UGT and GST specially responded to the pesticides. These findings will be useful in developing effective strategies for management of FAW in China and other invaded areas.
Animals
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China
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Genomics
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Humans
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Male
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Pesticides
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Spodoptera/genetics*
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Transcriptome