Study on the mechanism of Huanglian Jiedu Decoction in alleviating sepsis induced myocardial dysfunction in rats
Objective To explore the mechanism of Huanglian Jiedu Decoction in alleviating sepsis induced myocardial dysfunction(SIMD)in rats.Methods From 60 SPF grade SD male rats,20 rats were randomly selected and divided into long noncoding RNA nuclear-enriched abundant transcript 1(NEAT1)silencing group and the no-load group.From the remaining 40 rats,7 rats were selected as the sham operation group,and the remaining 33 rats were used to construct the sepsis model.In this experiment,27 rats were successfully modeled and randomly divided into the model group,the positive control group,and the Huanglian Jiedu Decoction group,with 9 rats in each group.At 28 days of modeling,the intervention was carried out,the sham operation group and the model group were given equal volumes of normal saline by gavage,the positive control group was given losartan potassium of 5 mg·kg-1 by gavage,and the NEAT1 silencing group,the no-load group and Huanglian Jiedu Decoction group were given Huanglian Jiedu Decoction of 12 mg·kg-1 by gavage.Myocardial histopathological changes were observed by HE staining.Long noncoding RNA nuclear-enriched abundant transcript 1(lncRNA-NEAT1)subcellular localization was detected by RNA FISH.The expression of inflammatory factors was detected by ELISA.The expression of lncRNA-NEAT1,microRNAs-132-3p(miR-132-3p)and uncoupling protein 2(UCP2)was detected by RT-PCR.Western blot assay was used to detect the expression of apoptosis and proliferation related proteins.Results After administration,compared with the model group,the disorder of myocardial fiber arrangement in Huanglian Jiedu Decoction group was relieved,and the swelling and necrosis of cardiomyocytes,and inflammatory cell infiltration were significantly improved.IncRNA-NEAT1 stimulated purplish red and was distributed in both the nucleus and cytoplasm of cardiomyocytes.Compared with the model group,microRNAs-132-3p levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β),the relative expression of lncRNA-NEATl mRNA,and the expression of Bax,Caspase-3 and Caspase-9 in the positive control group,Huanglian Jiedu Decoction group,the no-load group and NEAT1 silencing group were decreased(P＜0.05),with NEAT1 silencing group being the lowest;The relative expression levels of miR-132-3p and UCP2 mRNA and the expression of Ki67 were all increased(P＜0.05),with NEAT1 silencing group being the highest.Conclusion The mechanism which Huanglian Jiedu Decoction alleviates SIMD in rats may be that after downregulating lncRNA-NEAT1 and competitively binding to miR-132-3p,the inhibition of UCP2 is relieved,and after promoting UCP2 expression,the inflammatory response is further inhibited.
sepsiscardiac dysfunctionHuanglian Jiedu Decoctionanimal modelmechanism of action
万方数据
维普
