Effects of Polygonum cuspidatum on AMPK-FOXO3α Signaling Pathway in Rat Model of Uric Acid-Induced Renal Damage.

BACKGROUND: To observe the effects of Chinese medicine (CM) Polygonum cuspidatum (PC) on adenosine 5'-monophosphate-activated protein kinase (AMPK), forkhead box O3α (FOXO3α), Toll-like receptor-4 (TLR4), NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and monocyte chemoattractant protein-1 (MCP-1) expression in a rat model of uric acid-induced renal damage and to determine the molecular mechanism. METHODS: A rat model of uric acid-induced renal damage was established, and rats were randomly divided into a model group, a positive drug group, and high-, medium-, and low-dose PC groups (n=12 per group). A normal group (n=6) was used as the control. Rats in the normal and model groups were administered distilled water (10 mL•kg-1) by intragastric infusion. Rats in the positive drug group and the high-, medium-, and low-dose PC groups were administered allopurinol (23.33 mg•kg-1), and 7.46, 3.73, or 1.87 g•kg-1•d-1 PC by intragastric infusion, respectively for 6 to 8 weeks. After the intervention, reverse transcription polymerase chain reaction, Western blot, enzyme linked immunosorbent assay, and immunohistochemistry were used to detect AMPK, FOXO3α, TLR4, NLRP3, and MCP-1 mRNA and protein levels in renal tissue or serum. RESULTS: Compared with the normal group, the mRNA transcription levels of AMPK and FOXO3α in the model group were significantly down-regulated, and protein levels of AMPKα1, pAMPKα1 and FOXO3α were significantly down-regulated at the 6th and 8th weeks (P<0.01 or P<0.05). The mRNA transcription and protein levels of TLR4, NLRP3 and MCP-1 were significantly up-regulated (P<0.01 or P<0.05). Compared with the model group, at the 6th week, the mRNA transcription levels of AMPK in the high- and medium-dose groups, and protein expression levels of AMPKα1, pAMPKα1 and FOXO3α in the high-dose PC group, AMPKα1 and pAMPKα1 in the mediumdose PC group, and pAMPKα1 in the low-dose PC group were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription and protein levels of TLR4 and NLRP3 in the 3 CM groups, and protein expression levels of MCP-1 in the medium- and low-dose PC groups were down-regulated (P<0.01 or P<0.05). At the 8th week, the mRNA transcription levels of AMPK in the high-dose PC group and FOXO3α in the medium-dose PC group, and protein levels of AMPKα1, pAMPKα1 and FOXO3α in the 3 CM groups were significantly up-regulated (P<0.01 or P<0.05); the mRNA transcription levels of TLR4 in the medium- and low-dose PC groups, NLRP3 in the high- and low-dose PC groups and MCP-1 in the medium- and low-dose PC groups, and protein expression levels of TLR4, NLRP3 and MCP-1 in the 3 CM groups were down-regulated (P<0.01 or P<0.05). CONCLUSION: PC up-regulated the expression of AMPK and its downstream molecule FOXO3α and inhibited the biological activity of TLR4, NLRP3, and MCP-1, key signal molecules in the immunoinflammatory network pathway, which may be the molecular mechanism of PC to improve hyperuricemia-mediated immunoinflflammatory metabolic renal damage.

[Effects of Tengmei Decoction on the PPAR-y/NF-KB Signaling Pathway in Synovium of Collagen-in- duced Arthritis Rats].

Objective To observe the effects of Tengmei Decoction (TMD) on the expressions of peroxisome proliferator activated receptor gamma (PPARγ) , nuclear factor kappa B (NF-κB) , and IL- 17 in synovium of collagen-induced arthritis (CIA) rats, and to study its molecular mechanismpf. inhibi- ting synovial immune inflammatory injuries. Methods CIA model was established in Sprague-Dawley rats. Successfully modeled rats were randomly divided into the model group, the positive drug ,oup, high and low dose TMD groups, 6 in each group. Besides, a normal group was set up (n =6). Deionized water (10 mL . kg⁻¹ . d⁻¹) was administrated to rats in the normal group and the model group by gastro- gavage. Leflunomide (1. 87 mg . kg ⁻¹ . d ⁻¹) was administrated to rats in the positive drug group by gastro- gavage. TMD (31. 8 g crude drugs . kg ⁻¹ . d ⁻¹ and 15. 9 g crude drugs . kg ⁻¹ . d ⁻¹) was administrated to rats in high and low dose TMD groups respectively by gastrogavage. The intervention lasted for 12 suc- cessive weeks. Protein and mRNA levels of PPARy, P65, and IL-17 were detected at the end of intervention. Results Compared with the normal group, mRNA and protein expression levels of PPARγ, P65, and IL-17 were up-regulated in the model group (P <0. 01). Compared with the model group, PPARγ pro- tein expression level was up-regulated, mRNA and protein expression levels of P65 and IL-17 were down-regulated in high dose TMD group (P <0. 01). mRNA and protein expression levels of PPARγ were up-regulated, mRNA and protein expression levels of P65 and IL-17 were down-regulated in the positive drug group and low dose TMD group (P <0. 01). Conclusions TMD could ameliorate pathological damage of joint synovium , and inhibit expressions of immune inflammatory factors.

[Analysis of characteristics of using traditional Chinese and western medicine on type 1 diabetes adult patients based on electronic medical records].

To understand the kinds and the characteristics of combination of traditional Chinese medicines and western medicines using on adults with type 1 diabetes in general hospitals,This research brings into 4 602 hospitalized adult patients with type 1 diabetes from hospital information system (HIS) of 13 third class A hospitals. These research objects were hospitalized in December 2003-July 2011. The research analyzes the operating frequency and associated usage of western medicines and traditional Chinese medicines by the method of frequency statistics and association rules. Through the analysis, the research says that in the clinical treatment of adult patients with type 1 diabetes, the western medicine used most frequently is insulin, a total of 1 539 cases, accounted for 8.47%; the traditional Chinese medicine used most frequently is oral agents of pseudo-ginseng, a total of 183 cases, accounting for 6.25%; the combinations of Chinese and western medicines commonly used include Huoxue Huayu Tongmai Shuluo decoction + vasodilator, support degree is 45.93%, followed by Huoxue Huayu Tongmai Shuluo decoction + vasodilator + hypoglycemic drugs, support degree is 45.50%; Huoxue Huayu Tongmai Shuluo decoction + vasodilator + nutritional agent, support degree is 36.29%.

Upregulation of cystathionine-ß-synthetase expression contributes to inflammatory pain in rat temporomandibular joint.

BACKGROUND: Hydrogen sulfide (H2S), an endogenous gaseotransmitter/modulator, is becoming appreciated that it may be involved in a wide variety of processes including inflammation and nociception. However, the role for H2S in nociceptive processing in trigeminal ganglion (TG) neuron remains unknown. The aim of this study was designed to investigate whether endogenous H2S synthesizing enzyme cystathionine-ß-synthetase (CBS) plays a role in inflammatory pain in temporomandibular joint (TMJ). METHODS: TMJ inflammatory pain was induced by injection of complete Freund's adjuvant (CFA) into TMJ of adult male rats. Von Frey filaments were used to examine pain behavioral responses in rats following injection of CFA or normal saline (NS). Whole cell patch clamp recordings were employed on acutely isolated TG neurons from rats 2 days after CFA injection. Western blot analysis was carried out to measure protein expression in TGs. RESULTS: Injection of CFA into TMJ produced a time dependent hyperalgesia as evidenced by reduced escape threshold in rats responding to VFF stimulation. The reduced escape threshold was partially reversed by injection of O-(Carboxymethyl) hydroxylamine hemihydrochloride (AOAA), an inhibitor for CBS, in a dose-dependent manner. CFA injection led to a marked upregulation of CBS expression when compared with age-matched controls. CFA injection enhanced neuronal excitability as evidenced by depolarization of resting membrane potentials, reduction in rheobase, and an increase in number of action potentials evoked by 2 and 3 times rheobase current stimulation and by a ramp current stimulation of TG neurons innervating the TMJ area. CFA injection also led to a reduction of IK but not IA current density of TG neurons. Application of AOAA in TMJ area reduced the production of H2S in TGs and reversed the enhanced neural hyperexcitability and increased the IK currents of TG neurons. CONCLUSION: These data together with our previous report indicate that endogenous H2S generating enzyme CBS plays an important role in TMJ inflammation, which is likely mediated by inhibition of IK currents, thus identifying a specific molecular mechanism underlying pain and sensitization in TMJ inflammation.

Electroacupuncture suppresses mechanical allodynia and nuclear factor κ B signaling in streptozotocin-induced diabetic rats.

AIMS: To investigate whether electroacupuncture (EA) produced analgesic effect and whether nuclear factor kappa B (NF-κB) and cystathionine ß synthase (CBS) involved in EA-mediated analgesia in painful diabetic neuropathy in rats. METHODS: Diabetes was induced by an intraperitoneal injection of streptozotocin (STZ) in adult female rats. Mechanical pain threshold was measured by von Frey filaments. EA was applied at acupoint Zu-San-Li (ST-36) in both hindlimbs. Western blot analysis was employed to detect changes in protein levels of NF-κB and CBS in spinal dorsal root ganglion (DRGs). RESULTS: Mechanical allodynia was developed 2 weeks after STZ injection and lasted for another 4 weeks. STZ injection significantly enhanced expression of p65 and CBS in lumbar L4-6 DRGs when compared with age-matched controls. EA markedly attenuated mechanical allodynia. Importantly, EA treatment remarkably inhibited p65 and CBS expression in DRGs. Additionally, intrathecal injection of the p65 antagonist pyrrolidine dithiocarbamate attenuated mechanical allodynia and markedly inhibited CBS expression in DRGs in STZ rats. CONCLUSIONS: These data indicate that EA produced an analgesic effect, which might be mediated at least in a part by inhibition of NF-κB signaling pathway in primary sensory neurons in rats with diabetes.