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1.Effects of Didang Decoction-containing serum on high glucose-induced injury of rat glomerular endothelial cells

Bao-Lu LUO ; Quan-Gen CHU ; Jun CHU ; Fei-Xiang LI ; Jing CHEN ; Yue-Qi WANG

Chinese Traditional Patent Medicine 2024;46(9):2930-2935

2.Effect of Huangqi Guizhi Wuwutang on AGEs/RAGE/NF-κB Signaling Pathway in Diabetic Peripheral Neuropathy Rats

Ying FANG ; Ya-dong WANG ; Wen ZHOU ; Quan-gen CHU ; Wan-qiu HUANG

Chinese Journal of Experimental Traditional Medical Formulae 2020;26(13):52-58

Objective:To study the effect of Huangqi Guizhi Wuwutang on receptor of advanced glycation end products(AGEs)/advanced glycation endproducts (RAGE)/nuclear transcription factor-kappa B p65 (NF-κB p65) signaling pathway in the diabetic peripheral neuropathy rats through an animal modeling experiment, and discuss the mechanism of Huangqi Guizhi Wuwutang in alleviating diabetic peripheral neuropathy. Method:Rat model of diabetic peripheral neuropathy was established by high-fat diet and intraperitoneal injection with streptozotocin (STZ). After successful modeling, Huangqi Guizhi Wuwutang intervention began in the fifth week. The patients in high-dose group (19.40 g∙kg^-1∙d^-1), middle-dose group (4.85 g∙kg^-1∙d^-1) and low-dose group (2.43 g∙kg^-1∙d^-1) were given by gavage continuously for 12 weeks. The western medicine control group was given 25 mg∙kg^-1∙d^-1by gavage. After the experiment, serum interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect RAGE and NF-κB p65 mRNA expressions in sciatic nerve tissue. The expressions of RAGE, NF-κB and phosphorylation(p)-NF-κB p65 proteins in sciatic nerve tissues were detected by Western blot (WB). Result:Compared with the normal group, serum IL-1β and TNF-α protein levels, RAGE mRNA and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were significantly increased in the model group (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was increased, and the phosphorylation of NF-κB p65 was enhanced (P<0.01). After the intervention of Huangqi Guizhi Wuwutang, compared with the model group, serum IL-1β and TNF-α protein levels, RAGE and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were all decreased (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was decreased in high-dose group (P<0.01). The effect was obvious with the increase of dose of astragalus cassia twig. Conclusion:Huangqi Guizhi Wuwutang can alleviate diabetic peripheral neuropathy, and its mechanism may be related to blocking the expression of RAGE on tissue cell surface in AGEs/RAGE/NF-κB signaling pathway, inhibiting the activation of NF-κB and inducing TNF-α triggered oxidative stress and excessive inflammatory response, so as to avoid cell damage and dysfunction.

3.Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway

Fei-xiang LI ; Quan-gen CHU ; Pan WANG ; Jin YU ; Jun CHU

Chinese Journal of Experimental Traditional Medical Formulae 2021;27(22):59-64

Objective:To observe the intervention of phlegm-stasis co-treatment on the myocardial Toll-like receptor 4 （TLR4）/nuclear factor-κB （NF-κB）/nuclear factor-κB inhibitor （IκB） signaling pathway， and to investigate its mechanism in improving myocardial inflammation in rats with diabetes mellitus （DM）. Method:Forty-five male SD rats of SPF grade were randomly divided into a normal group， a phlegm-resolving （Xiao Xianxiongtang， 4.05 g·kg^-1） group， a stasis-resolving （Xuefu Zhuyutang， 7.02 g·kg^-1） group， a co-treatment （Didang Xianxiong decoction， 8.10 g·kg^-1） group， an alagebrium chloride （3 mg·kg^-1） group， and a model group. Except for normal group， the other rats was induced by a single intraperitoneal injection of 55 mg·kg^-1streptozotocin （STZ） to establish DM model. After adaptive feeding for three weeks， the rats were treated correspondingly by gavage daily for eight weeks. Rats were sampled under anesthesia. Enzyme-linked immunosorbent assay（ELISA） was used to detect the protein expression of TLR4 and tumor necrosis factor-alpha （TNF-α） in myocardial tissues. The expression levels of NF-κB p65 and IκBα were detected by immunohistochemistry. NF-κB p65， IκBα， TNF-α， and TLR4 mRNA expression levels were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）. Result:The protein and mRNA levels of TLR4， NF-κB p65， IκBα， and TNF-α were higher in the model group than those in the normal group （P<0.01）. TLR4， NF-κB p65， IκBα， and TNF-α protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group （P<0.01）. The inter-group comparison revealed that the co-treatment group showed more manifest reduction in protein and mRNA expression levels of TLR4， NF-κB p65， IκBα， and TNF-α than the phlegm-resolving group and the stasis-resolving group （P<0.05，P<0.01）. Conclusion:The co-treatment of phlegm and stasis can improve myocardial inflammation in DM rats， with superior effect to either the phlegm-resolving method or the stasis-resolving method. The underlying mechanism may be related to the inhibition of TLR4/NF-κB/IκB signaling pathway activation.

4.Effect of Didang Xianxiong Decoction on Cardiac Myocardial Microvascular Endothelial Cell Injury Induced by High Glucose

Jin YU ; Quan-gen CHU ; Fei-xiang LI ; Zheng-yin CAI ; Jun CHU

Chinese Journal of Experimental Traditional Medical Formulae 2022;28(9):52-58

ObjectiveThis study was designed to observe the effect of Didang Xianxiong decoction on the cardiac myocardial microvascular endothelial cells （CMECs） injury， and to explore its related mechanism based on the CMECs model induced by high glucose. MethodRat primary myocardial cells were cultured in vitro and 33 mmol·L^-1 glucose was added for modeling. After modeling， the rats were randomly divided into model group （final glucose concentration： 33 mmol·L^-1）， normal group， Didang Xianxiong decoction low dose group （glucose + 5% Didang Xianxiong decoction containing serum）， Didang Xianxiong decoction medium dose group （glucose+10% Didang Xianxiong decoction containing serum）， Didang Xianxiong decoction high dose group （glucose+20% Didang Xianxiong decoction containing serum） and alagebrium chloride （ALT-711） group （glucose+10% ALT-711 containing serum）. The influence of drug-containing serum on the proliferation of CMECs was detected by MTT tetrazolium salt colorimetric assay. The relative mRNA expression of c-Jun was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein expression of phosphorylated Janus kinase 1 （p-JAK1）， phosphorylated signal transducer and activator of transcription 1 （p-STAT1） and transforming growth factor-β₁ （TGF-β₁） was determined by Western blot. ResultCompared with the conditions in normal group， the mRNA expression of c-Jun and protein expression of p-JAK1， p-STAT1 and TGF-β₁ were up-regulated in model group （P<0.01）. Compared with model group， all treatment groups had decreased mRNA expression of c-Jun （P<0.01）. Didang Xianxiong decoction medium and high dose groups and ALT-711 group showed reduced protein expression of p-JAK1 and p-STAT1 （P<0.05， P<0.01）， while there was no significant change in Didang Xianxiong decoction low dose group. TGF-β₁ protein expression was lowered in all treatment groups （P<0.05， P<0.01）， and the decrease was more significant in Didang Xianxiong decoction medium and high dose groups than Didang Xianxiong decoction low dose group. ConclusionDidang Xianxiong decoction can protect CMECs with high glucose-induced injury， and the mechanism may be related to reducing the activity of JAK/STAT signaling pathway in cells.

1.Effects of Didang Decoction-containing serum on high glucose-induced injury of rat glomerular endothelial cells

2.Effect of Huangqi Guizhi Wuwutang on AGEs/RAGE/NF-κB Signaling Pathway in Diabetic Peripheral Neuropathy Rats

3.Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway

4.Effect of Didang Xianxiong Decoction on Cardiac Myocardial Microvascular Endothelial Cell Injury Induced by High Glucose

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