Effect of Didang Xianxiong Decoction on Cardiac Myocardial Microvascular Endothelial Cell Injury Induced by High Glucose / 中国实验方剂学杂志

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.

Co-Treatment of Phlegm and Stasis Improves Myocardial Inflammation in Rats with Diabetes Mellitus via TLR4/NF-κB/IκB Pathway / 中国实验方剂学杂志

Objective:To observe the intervention of phlegm-stasis co-treatment on the myocardial Toll-like receptor 4 （TLR4）/nuclear factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）/nuclear factor-<italic>κ</italic>B inhibitor （I<italic>κ</italic>B） 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-<italic>α</italic>） in myocardial tissues. The expression levels of NF-<italic>κ</italic>B p65 and I<italic>κ</italic>B<italic>α</italic> were detected by immunohistochemistry. NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， TNF-<italic>α</italic>， 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-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α </italic>were higher in the model group than those in the normal group （<italic>P</italic><0.01）. TLR4， NF-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α</italic>， and TNF-<italic>α</italic> protein and mRNA expression levels were reduced to varying degrees in the groups with drug intervention as compared with those in the model group （<italic>P</italic><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-<italic>κ</italic>B p65， I<italic>κ</italic>B<italic>α，</italic> and TNF-<italic>α </italic>than the phlegm-resolving group and the stasis-resolving group （<italic>P</italic><0.05<italic>，P</italic><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-<italic>κ</italic>B/I<italic>κ</italic>B signaling pathway activation.