1.Mechanism of Modified Tianwang Buxindan on Skin of Sleep-deprived Mice Through PI3K/Akt/Nrf2 Signaling Pathway
Juanping CHEN ; Yuan PENG ; Xuemin HONG ; Li YANG ; Bo XU ; Chong ZHANG ; Xuelin GUO
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(11):120-128
ObjectiveTo observe the effect of modified Tianwang Buxindan (MTBD) on the skin of sleep-deprived (SD) mice and investigate its mechanism. MethodSixty 2-month-old female Kunming mice were randomly divided into a blank group, a model group, a vitamin C (VC, 0.08 g·kg-1), and MTBD low-, medium-, and high-dose groups (6.5, 12.5, 25 g·kg-1). Except for the blank group, the other groups were subjected to SD mouse model induction (using multiple platform water environment method for 18 hours of sleep deprivation daily from 15:00 to next day 9:00), continuously for 14 days, and caffeine (CAF, 7.5 mg·kg-1) was injected intraperitoneally from the 2nd week onwards, continuously for 7 days. While modeling, the blank group and the model group were administered with normal saline (0.01 mL·g-1), and the other groups received corresponding drugs for treatment. On the day of the experiment, general observations were recorded (such as body weight, spirit, fur, and skin). After sampling, skin tissue pathological changes were observed under an optical microscope using hematoxylin-eosin (HE) and Masson staining methods. Skin thickness and skin moisture content were measured. Biochemical assay kits were used to detect skin hydroxyproline (HYP) content, skin and serum superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum interleukin (IL)-6, tumor necrosis factor (TNF)-α, and IL-1β levels in mice. Western blot was used to detect skin tissue type Ⅰ collagen (ColⅠ), type Ⅲ collagen (ColⅢ), phosphatidylinositol 3-kinase (PI3K), phosphorylated (p)-PI3K, protein kinase B (Akt), p-Akt, nuclear factor E2-related factor 2 (Nrf2), heme oxygenase (HO)-1, and nuclear factor (NF)-κB protein expression. ResultCompared with the blank group, the model group showed varying degrees of changes. In general, signs of aging such as reduced body weight (P<0.01), listlessness, dull fur color, and formation of wrinkles on the skin appeared. Tissue specimen testing revealed skin thinning, flattening of the dermoepidermal junction (DEJ), and reduced collagen fibers under the optical microscope. Skin thickness and moisture content decreased, skin tissue HYP content significantly decreased (P<0.01), skin and serum SOD activity significantly decreased (P<0.01), and MDA content significantly increased (P<0.01). Serum IL-6, TNF-α, and IL-1β levels significantly increased (P<0.01). Skin ColⅠ, ColⅢ, p-PI3K/PI3K, p-Akt/Akt, Nrf2, and HO-1 protein expression significantly decreased (P<0.05, P<0.01), and NF-κB expression increased (P<0.01). Compared with the model group, the VC group and the MTBD low-dose group showed increased skin moisture content, HYP content, SOD activity, and ColⅠ, ColⅢ, p-PI3K/PI3K protein expression (P<0.05, P<0.01), and decreased serum MDA content (P<0.05). In addition, a decrease in serum IL-6 and IL-1β levels was detected in the MTBD low-dose group (P<0.05), while the above indicators in the MTBD medium- and high-dose groups improved (P<0.05, P<0.01). ConclusionSleep deprivation accelerates the aging process of the skin in SD model mice. MTBD can improve this phenomenon, exerting anti-inflammatory and antioxidant effects, and its mechanism of action may be related to the activation of the PI3K/Akt/Nrf2 signaling pathway.
2.Health risk assessment of chemical pollutants in drinking water in Nanjing from 2014 to 2022
Yuhong ZHENG ; Jingshan ZHANG ; Chunjing CHEN ; Ming GE
Journal of Environmental and Occupational Medicine 2024;41(8):905-910
Background Drinking water contains a variety of chemicals that may pose certain health risks to the human body. Objective To evaluate carcinogenic and non-carcinogenic health risks of chemical pollutants in drinking water in Nanjing from 2014 to 2022. Methods According to the Standard examination methods for drinking water (GB/T 5750-2006) and the Standards for drinking water quality (GB 5749-2006), the conventional water quality indexes of finished water, tap water, and secondary water supply in Nanjing from 2014 to 2022 were monitored. The health risk assessment model recommended by the United States Environmental Protection Agency (US EPA) was used to assess the health risks of 16 chemicals [arsenic, cadmium, chromium (hexavalent), lead, mercury, selenium, cyanide, fluoride, nitrate nitrogen, trichloromethane, carbon tetrachloride, aluminum, iron, manganese, copper, and zinc] in drinking water through different routes (drinking water and skin contact) in different populations (adult males, adult females, and children). Region (urban and rural), water period (dry period and wet period), and water sample type (finished water, tap water, and secondary water supply) were stratified for analysis. Results From 2014 to 2022, a total of
3.Determination of camphor residue and borneol content in Qingchang suppository by GC
Yujiong PAN ; Zhigao HE ; Xin ZHOU ; Hengzhou ZHANG ; Yuehong YANG ; Jingshan HUANG
Journal of Pharmaceutical Practice 2023;41(9):552-556
Objective To establish a gas chromatography for simultaneous determination of camphor residue and borneolum content in Qingchang Suppository. Methods Gas chromatograph method was used. The chromatographic column was Agilent capillary column(30 m×0.25 mm×0.25 µm). The column temperature was 140 ℃. The sample injection temperature was 250 ℃. The FID detector temperature was 250 ℃. Results Camphor,borneol and isoborneol content showed good linear in the extent of 0.0299~1.497(r=1.000), 0.0205~1.025(r=1.000), 0.0097~0.4830 µg (r=1.000). RSDs of precision,stability and repeatability test results were less than 2%. The recovery was 99.7%, 101.0%, 102.5%. Conclusion This method is simple and quick with accurate result, which could be used for the content determination of Borneol in Qingchang Suppository.
4.Alterations of gut microbiota composition and cogni-tive behavior in APP/PS1/tau triple-transgenic mouse model of Alzheimer's disease
Chinese Journal of Pharmacology and Toxicology 2023;37(7):521-521
OBJECTIVE Alzheimer's disease(AD)is a progressive neurological disease.Given the important role of gut microbiota composition in AD pathology,the observed perturbation in the microbiota composition and diversity may serve as the mechanisms underlying age-dependent APP/PS1/tau triple-transgenic mouse(3×Tg-AD)mice amyloid deposition and memory deficits.Here-in,we intended to investigate the gut microbiota and as-sessed its relationship with the triggering and develop-ment of cognitive impairment of AD.METHODS This study involves the comparative assessment of spatial learning,amyloid β-protein(Aβ)accumulation,and fecal microbiota alterations in 3×Tg-AD mice from three age groups:AD asymptomatic stage(3 m),presymptomatic stage(6 m),and the symptomatic stage of AD(9 m).RE-SULTS We demonstrate that spatial memory deficits,brain Aβ accumulation,and weight gain in 3×Tg-AD mice gradually appear after 6 months of age.However,the total gut bacterial counts underwent changes from 3 to 6 months of age and were further altered at 9 months of age.Importantly,changes in gut bacteria abundance of Desulfobacterota and Actinobacteriota phylain 6-month-old mice preceded apparent spatial memory deficits.CONCLUSION Changes in the gut microbial community are one of the mechanisms of early AD pathology.
5. Cloning and expressive characteristics of GPS gene and its correlation with saponins content in Eleutherococcus senticosus
Chinese Traditional and Herbal Drugs 2019;50(5):1227-1231
Objective To clone cDNA sequence of geranyl pyrophosphate synthase (GPS) gene from Eleutherococcus senticosus and analyze genetic characteristics, gene expression level in different organs and the correlation between GPS gene expression and saponins content. Methods RNA was extracted from E. senticosus and reverse transcribed into cDNA. Gene specific primers were designed according to the unigene (c37362.graph_c0) of GPS from transcriptome sequencing data. The full length of the GPS gene cDNA was amplified by PCR. The expression level of GPS gene in different organs was analyzed by qRT-PCR. The content of E. senticosus saponins was detected by spectrophotometry method. Results GPS gene cDNA was cloned from E. senticosus and encodes 419 amino acids with full length of 1 260 bp. GPS protein located in mitochondria does not have transmembrane region. The GPS gene was expressed in each organ and had the highest expression in blade, which is 5.26 times in root. The relative expression of GPS gene and content of saponin showed the same trend and significant positive correlation (r = 0.851, P < 0.05). Conclusion The whole length of cDNA sequence of GPS gene is cloned for the first time, and there is a positive correlation between the expression level of GPS gene and the saponin content of E. senticosus.
6.The preliminary research of T2 mapping at 3.0T MR in lumbar intervertebral disc degeneration
Jingshan TAO ; Xianhua WU ; Feng HAN ; Juan ZHANG ; Yingdi XIE
Journal of Practical Radiology 2016;(2):288-291
Objective The purpose of our research was to quantitative study the degeneration of lumbar intervertebral discs by using T2 mapping technology at 3.0T MR and research the correlation of biomechanics and lumbar disc degeneration.Methods 34 patients with low back pain or sciatica were enrolled.The correlation of T2 values of lumbar intervertebral discs and Pfirrmann grading,disc location,patient ages were analyzed.T2 values before and after loading were also analyzed.Results T2 values of nucleus pulposus were correlation with Pfirrmann scoring,disc location and,patient ages.Conclusion (1 )T2 mapping provides a non-invasive and quantitative way to diagnose lumbar discs degeneration.(2)The change of external pressure load can affect the T2 values of the intervertebral disc.
7.Protective effects of NDP on LPS induced neuron injuries in rat mixed cultures
Mu LIN ; Qihai GONG ; Qin WU ; Feng ZHANG ; Jingshan SHI
Chinese Pharmacological Bulletin 2016;32(8):1144-1147,1148
Aim To investigate the protective effect of noble dendrobium polysaccharides ( NDP ) on lipopo-lysaccharide ( LPS)-induced neuron injuries in newborn rat cerebral cortex glial cells and neuron mixed cul-tures.Methods The primary cultures of newborn rat cortical neurons and glial cells were established and the existence of the neurons , astrocytes and microglia was verified respectively .NDP was given to LPS-induced mixed cultures , the mRNA levels of IL-1β, TNF-αand COX-2 were assayed by real time PCR .Results NDP reduced the glial cell activation and neuron dam-age after it was given to LPS-induced mixed cultures . The mRNA levels of IL-1β, TNF-α, COX-2 were re-duced .Conclusion NDP protects against LPS-in-duced neuron-inflammation in neurons and glial cells cultures.
8.Hydroxysafflor yellow A inhibits VSMCs proliferation via PCNA and MEK-ERK1/2
Jingshan ZHAO ; Mingxing FANG ; Qianyu GUO ; Yunfeng LI ; Bingyuan XU ; Shaohong LAI ; Shangyue ZHANG ; Yu LIU ; Aiying LI
Chinese Pharmacological Bulletin 2015;(7):984-987,988
Abstrac:Aim To study the effect of hydroxysafflor yellow A ( HYSA ) on the proliferation of vascular smooth muscle cells ( VSMCs) and the related molecu-lar mechanism. Methods The inhibitory effects of hydroxysafflor yellow A on VSMC proliferation was de-tected using cell culture, MTT assay, Western blot and immunohistochemical staining. Results The results showed that HYSA inhibited cell proliferation induced by PDGF in a dose-dependent (5,10,20,40 μmol· L-1 ) manner, reduced proliferating cell nuclear anti-gen ( PCNA ) expression and blocked PDGFR-MEK-ERK1/2 signaling pathway activated by PDGF in VSMCs. Conclusion HYSA inhibits VSMCs prolifer-ation via reducing the expression of PCNA and blocking signal transduction of MEK-ERK1/2 in VSMCs.
9.Hydroxysafflor yellow A inhibits rat vascular smooth muscle cells proliferation possibly via blocking signal transduction of MEK-ERK1/2
Jingshan ZHAO ; Qianyu GUO ; Shaohong LAI ; Shangyue ZHANG ; Hongli HAN
Chinese Journal of Cardiology 2015;43(8):728-731
Objective To elucidate the effect of hydroxysafflor yellow A (HYSA) on the proliferation of vascular smooth muscle cells (VSMCs) and the related mechanism.Methods VSMCs derived from SD rats were treated with DMEC culture medium (Control),10 ng/ml PDGF (PDGF group),pretreatment with HYSA at different doses (1,5,10,20,40,60 μmol/L) for 24 h then cotreatment with PDGF.After 24 h,MTT assay,Western blot and immunohistochemical staining were performed to evaluate the inhibitory effects of HYSA on VSMCs proliferation.Results HYSA inhibited PDGF induced VSMCs proliferation in a dose-dependent manner,dowregulated proliferating cell nuclear antigen (PCNA) expression and blocked PDGF activated PDGFR-MEK-ERK1/2 signaling pathway.Conclusions HYSA inhibits VSMCs proliferation possibly via downregulating the expression of PCNA and blocking MEK-ERK1/2 signal transduction in VSMCs.
10.The effects of tetrahydroxystilbene glucoside on mouse liver cytochrome P 450
Feng ZHANG ; Hang LIU ; Yanying WANG ; Jing NIE ; Jingshan SHI
Chongqing Medicine 2013;(36):4418-4420
Objective To investigate the effects of tetrahydroxystilbene glucoside (TSG) on cytochrome P450(CYP) in mouse livers .Methods Kunming male mice were divided into the blank ,low dose and the high dose of TSG groups .3 ,5 and 7 after intra-gastrical administration of TSG ,mice were sacrificed and the mRNA expressions of CYP isoenzymes in mouse livers were measured by real time reverse transcription-polymerase chain reaction(RT-PCR) ,respectively .Results TSG significantly inhibited CYP1A2 and CYP 3A4 mRNA expressions at 3th ,5th and 7th day after treatment .TSG time-dependently increased CYP2E1 mRNA expres-sion .TSG inhibited CYP4A14 mRNA expression at 7th day after treatment .Moreover ,TSG had no significant effects on CYP2B10 , CYP3A11 and CYP3A25 mRNA expressions .Conclusion TSG has significant effects on CYP1A2 ,CYP2E1 ,CYP3A4 and CYP4A14 mRNA expressions but no significant effects on CYP2B10 ,CYP3A11 and CYP3A25 mRNA expressions .

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