Nuanxinkang protects against ischemia/reperfusion-induced heart failure through regulating IKKß/IκBα/NF-κB-mediated macrophage polarization.

BACKGROUND: Heart failure (HF) is a leading cause of death worldwide. Nuanxinkang (NXK) is an effective Chinese herbal formula used in treating HF, but its underlying potential mechanisms have not been fully elucidated. PURPOSE: To explore the protective activities of NXK in ischemia/reperfusion (IR)-induced HF through modulating the ratio of proinflammatory (M1) and anti-inflammatory (M2) macrophage populations and leading to the alleviation of inflammation. MATERIALS AND METHODS: In vivo, mice were subjected to myocardial IR to generate HF mouse models. Mice in the NXK group were treated with NXK for 28 days. Cardiac function was detected by echocardiography. Major lesions on mouse hearts were determined by hematoxylin-eosin (HE) staining, Masson staining, and TUNEL staining. Inflammatory cytokines were determined by enzyme-linked immunosorbent assay (ELISA) and qPCR examination. Flow cytometric analyses and qPCR examination were utilized for monitoring the temporal dynamics of macrophage infiltration following IR. In vitro, two polarized models were established by stimulating RAW264.7 cells with 200 ng/ml lipopolysaccharide (LPS) or 20 ng/ml interleukin-4 (IL-4). The RAW264.7 cells with nuclear factor-κB (NF-κB) overexpression was generated by transient transfection of NF-κB plasmids, and NXK intervention was conducted on this cell model to further clarify the involvement of NF-κB signaling in the NXK-mediated HF process. RESULTS: In the present study, NXK was found to significantly contribute the cardiac function and ameliorate cardiac fibrosis and apoptosis after myocardial IR injury in vivo, which may be partially due to a decrease in inflammation. We therefore hypothesized that NXK reduced inflammatory damage by modulating subtypes of macrophages. And the results demonstrated that the percentage of proinflammatory macrophages infiltrated in the post-IR period was reduced with NXK treatment, and thereby blunting the wave of proinflammatory response and shifting the peak of the anti-inflammatory macrophage-mediated wound healing process towards an earlier time point. The further investigation showed that macrophage polarization was mediated by NXK through inhibiting the phosphorylation and the nuclear translocation of NF-κB. Besides, the phosphorylated IKKß and IκBα, upstream mediators of the NF-κB pathway, also decreased by NXK. Moreover, the overexpression of NF-κB partially reversed the NXK-induced favorable activities; and successfully compensated the suppressive effect on inflammation and the phosphorylation of NF-κB. CONCLUSION: In conclude, our results demonstrated that NXK induced the cardioprotective effects against IR injury through a regulatory axis of IKKß/IκBα/NF-κB-mediated macrophage polarization. The information gained from this study provide a possible natural strategy for anti-inflammatory treatment of HF.

[Rule of acupoint selection for cancer pain based on association rule and entropy clustering].

OBJECTIVE: To analyze the rule of acupoint selection for cancer pain based on data mining. METHODS: The published literature regarding acupuncture for cancer pain in the recent 10 years was searched in PubMed, CNKI, VIP and Wanfang database. The acupoint selection was summarized and analyzed by TCMISS V2.5. RESULTS: Totally 68 literature was collected and 73 acupoint prescriptions were included, involving 117 acupoints. These acupoints were mainly in bladder meridian, stomach meridian, liver meridian and spleen meridian. Among them, 40 acupoints used more than 4 times were identified, and the top three acupoints were Zusanli (ST 36, 65 times), Neiguan (PC 6, 55 times) and Taichong (LR 3, 50 times). A total of 68 acupoint combinations used more than 19 times were identified, and the top three acupoint combinations were Zusanli (ST 36)-Neiguan (PC 6), Taichong (LR 3)-Zusanli (ST 36) and Zusanli (ST 36)-Sanyinjiao (SP 6). There were 103 acupoint combinations with strong association; based on the entropy clustering algorithm, 20 new acupoint combinations and 10 new acupoint prescriptions were obtained. CONCLUSION: The main meridians for cancer pain are bladder meridian, stomach meridian, liver meridian and spleen meridian, with Zusanli (ST 36), Neiguan (PC 6), Taichong (LR 3), Hegu (LI 4), Sanyinjiao (SP 6) and ashi points as core acupoints, and regulating spleen-stomach and treating qi-blood are the main principles.

[Effect of Jiangtang Xiaozhi Tablet on Insulin Resistance KK-AY Transgenic Mice Model of Diabetes Mellitus].

Objective To observe effects of Jiangtang Xiaozhi Tablet (JTXZT) on homeostasis model of assessment for insulin resistance index (HOMA-IR) , insulin sensitivity index ( ISI) , expres- sions of insulin (INS) and insulin receptor (InsR) in pancreas tissues of KK-A(y) transgenic mice model of diabetes mellitus (DM). Methods KK-A(y) transgenic mice were fed with high fat forage to induce hyper- glycemic obese DM model. The C,7ice at same age were used as a normal control group (fed with e- qual volume of sterile water, n =11). Successful modeled 55 mice with DM obesity were divided into 5 groups by random digit table (11 in each group) , including the model group (fed with equal volume of ster- ile water, with no treatment) , the Pioglitazone Hydrochloride Tablet treatment group (8 mg/kg; as a posi- tive control group) , and JTXZT groups [high (10. 0 g crude drugs/kg) , middle (5. 0 g crude drugs/kg) and low dose (2. 5 g crude drugs/kg) ]. All medications were fed by gastrogavage, once per day for 8 succes- sive weeks. All mice were weighed and levels of random blood glucose (RBG) determined after 8 weeks of treatment. Blood was collected from ophthalmic vein. Levels of insulin (INS) , serum total cholesterol (TC) and triglyceride (TG) were detected. HOMA-IR and ISI were calculated. The morphological changes of pancreas tissues were extracted for performed pathological examinations. The expressions of INS and insulin receptor (InsR ) were measured by immunohistochemistry ( IHC ). Expressions of insulin receptorp ßInsRP) and insulin receptor substrate-1 (IRS-1) in pancreas tissues were detected using Western blot. Results Compared with the normal control group, obesity, obviously increased blood glu- cose and blood lipids occurred in each group after modeling (P <0. 01). After 8 weeks of medication mice in the model group had put up body weight (P <0. 01) , blood glucose and blood lipids were kept on quite higher levels. Compared with the model group, body weight, serum levels of TG, INS, and HOMA-IR obvi- ously decreased in each JTXZT group (P <0. 05, P <0. 01). Besides, RBG decreased obviously lower in the high dose JTXZT group (P <0. 01). ISI obviously increased in low and high dose JTXZT groups (P < 0. 05, P <0. 01). Pathological results of HE staining in pancreas showed that atrophied islets with obvious- ly reduced numbers in the model group. They were sparsely distributed with reduced islet density.-Islet cells were compensatively hypertrophy, with degenerated vacuoles. Apoptosis of islet cells could also be seen in the model group, manifested as swollen cytoplasm and paryopyknosis. Islet number was obvious- ly increased in high and middle dose JTXZT groups, with reduced apoptosis and degenerated cells. Re- sults of IHC assay showed, as compared with the normal control group, the grey values of INS and InsR were significantly decreased in the model group (P <0. 01). Compared with the model group, IOD values of INS and InsR (IOD) were significantly increased in each JTXZT group (P <0. 05, P <0. 01). Results from Western blot showed that protein expressions of InsRP ßnd IRS-1 were obviously decreased in the model group, as compared with the normal control group (P <0. 01). Compared with the model group, protein expressions of InsRP ßnd IRS-1 were obviously increased in each JTXZT group (P <0. 01) , but with no statistical difference as compared with the Pioglitazone Hydrochloride Tablet treatment group (P > 0. 05). Conclusions JTXZT had obvious roles in decreasing levels of blood glucose, serum lipids, and improving insulin resistance in KK-Ayt(r) ansgenic mice model with diabetic obesity. Its mechanism might involve in increasing expressions of lnsRp and IRS-1 in pancreas cells, promoting the integration of INS to its receptors, and thereby improving glucose metabolism , lipid metabolism , and IR state.