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

ABSTRACT

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.