[Sagittaria sagittifolia polysaccharides regulates Nrf2/HO-1 to relieve liver injury caused by multiple heavy metals in vivo and in vitro].

This study explored whether Sagittaria sagittifolia polysaccharides(SSP) activates the nuclear factor erythroid-2-related factor2(Nrf2)/heme oxygenase-1(HO-1) signaling pathway to protect against liver damage jointly induced by multiple heavy metals. First, based on the proportion of dietary intake of six heavy metals in rice available in Beijing market, a heavy metal mixture was prepared for inducing mouse liver injury and HepG2 cell injury. Forty male Kunming mice were divided into five groups: control group, model group, glutathione positive control group, and low-and high-dose SSP groups, with eight mice in each group. After 30 days of intragastric administration, the liver injury in mice was observed by HE staining. In the in vitro experiment, MTT assay was conducted to detect the effects of SSP at 0.25, 0.5, 1, and 2 mg·mL~(-1) on HepG2 cell survival at different time points. The content of alanine transaminase(ALT) and aspartate aminotransferase(AST) in the 48-h cell culture fluid was measured using micro-plate cultivation method, followed by the detection of the change in reactive oxygen species(ROS) content by flow cytometry. The mRNA expression levels of Nrf2 and HO-1 in cells were determined by RT-PCR, and their protein expression by Western blot. HE staining results showed that compared with the model group, the SSP administration groups exhibited significantly alleviated inflammatory cell infiltration and fatty infiltration in the liver, with better outcomes observed in the high-dose SSP group. In the in vitro MTT assay, compared with the model group, SSP at four concentrations all significantly increased the cell survival rate, decreased the ALT, AST, and ROS content(P<0.05), and down-regulated Nrf2 and HO-1 mRNA and protein expression(P<0.05). SSP significantly improves inflammatory infiltration in the liver tissue of mice exposed to a variety of heavy metals and corrects the liver fat degeneration, which may be related to its regulation of the Nrf2/HO-1 signaling pathway, reduction of ROS, and alleviation of oxidative damage.

Effect of electroacupuncture on glial fibrillary acidic protein and nerve growth factor in the hippocampus of rats with hyperlipidemia and middle cerebral artery thrombus.

Electroacupuncture (EA) has been shown to reduce blood lipid level and improve cerebral ischemia in rats with hyperlipemia complicated by cerebral ischemia. However, there are few studies on the results and mechanism of the effect of EA in reducing blood lipid level or promoting neural repair after stroke in hyperlipidemic subjects. In this study, EA was applied to a rat model of hyperlipidemia and middle cerebral artery thrombosis and the condition of neurons and astrocytes after hippocampal injury was assessed. Except for the normal group, rats in other groups were fed a high-fat diet throughout the whole experiment. Hyperlipidemia models were established in rats fed a high-fat diet for 6 weeks. Middle cerebral artery thrombus models were induced by pasting 50% FeCl3 filter paper on the left middle cerebral artery for 20 minutes on day 50 as the model group. EA1 group rats received EA at bilateral ST40 (Fenglong) for 7 days before the thrombosis. Rats in the EA1 and EA2 groups received EA at GV20 (Baihui) and bilateral ST40 for 14 days after model establishment. Neuronal health was assessed by hematoxylin-eosin staining in the brain. Hyperlipidemia was assessed by biochemical methods that measured total cholesterol, triglyceride, low-density lipoprotein and high-density lipoprotein in blood sera. Behavioral analysis was used to confirm the establishment of the model. Immunohistochemical methods were used to detect the expression of glial fibrillary acidic protein and nerve growth factor in the hippocampal CA1 region. The results demonstrated that, compared with the model group, blood lipid levels significantly decreased, glial fibrillary acidic protein immunoreactivity was significantly weakened and nerve growth factor immunoreactivity was significantly enhanced in the EA1 and EA2 groups. The repair effect was superior in the EA1 group than in the EA2 group. These findings confirm that EA can reduce blood lipid, inhibit glial fibrillary acidic protein expression and promote nerve growth factor expression in the hippocampal CA1 region after hyperlipidemia and middle cerebral artery thrombosis. All experimental procedures and protocols were approved by the Animal Use and Management Committee of Beijing University of Chinese Medicine, China (approval No. BUCM-3-2018022802-1002) on April 12, 2018.

[Effect of acupuncture at different layers of local "Neiguan" (PC6) tissue on arrhythmia and expression of myocardial Cx43 in rabbits].

OBJECTIVE: To compare the therapeutic effect of shallow and deep needling at "Neiguan"(PC6) in the treatment of arrhythmia in rabbits. METHODS: Male New Zealand rabbits were randomly divided into saline (n＝15), model (n＝12), shallow needling (n＝13) and deep needling (n＝12) groups. The arrhythmia model was established by ear intravenous injection of Barium chloride (0.4%, 1 mL/kg). Acupuncture needle was inserted into the superficial fascia (about 3 mm beneath the skin) or deep layer (5 to 8 mm, near the median nerve) of local PC6 tissue (fore limb), manipulated for a while and then retained for 10 min. Histopathological changes of myocardium was observed after H．E. stain, and the immunoactivity of connexin 43 (Cx43) detected by immunohistochemistry(IHC). RESULTS: Various degrees of arrhythmia and down-regulated expression of myocardial Cx43 were observed in all rabbits after modeling. After EA intervention and compared with the model group, the initial time of arrhythmia and Cx43 expression were obviously increased (P<0.01), and the duration of arrhythmia was significantly shortened in both deep and shallow needling groups (P<0.01). Compared with the shallow needling group, the Cx43 expression was increased in the deep needling group (P<0.01). H．E. staining showed disordered and wavy arrangement of myocardial fibers, with exudation of serous and erythrocytes in the myocardial interstitium in the model group, which was relatively mild in both needling groups. IHC showed disordered distribution of Cx43 in the ventricular myocytes and almost no obvious band-like distribution at the discs in rabbits of the model group, and abundant distribution of Cx43 in the sacral disc in the deep needling group, and strip-shaped, cluster-like, point-like, visible at both end-to-end connections and side-to-side connections in the shallow needling group. CONCLUSION: Both shallow and deep needling can significantly reduce the duration of arrhythmia in arrhythmia rabbits, which may be associated with its effect in up-regulating expression of myocardial Cx43 protein.