Pudilan xiaoyan oral liquid alleviates LPS-induced respiratory injury through decreasing nitroxidative stress and blocking TLR4 activation along with NF-ΚB phosphorylation in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Pudilan xiaoyan oral liquid (PDL), collected in Chinese Pharmacopoeia, has been used clinically for treating inflammatory diseases such as upper respiratory tract infection diseases. However, its potential anti-inflammation and the mechanism are still unclear. MATERIALS AND METHODS: lipopolysaccharide (LPS) was used to induce respiratory inflammation of mice by intratracheal administration. UPLC/MS was performed for components analysis of PDL. Enzyme-linked immune sorbent assay (ELISA) was conducted for determining interleukin-6(IL-6), interleukin-1ß(IL-1ß) and tumor necrosis factor-α(TNF-α) in serum and supernatant of tracheal tissue while Nitric oxide assay kit for nitric oxide (NO) content. Hematoxylin-Eosin (HE) staining was applied to evaluate pathological lesions. Western blotting analysis (WB) and Immunohistochemistry(IHC) were employed for the determination of Toll-like receptors 4(TLR4), TNF-α, IL-6, inducible nitric oxide synthase(iNOS) and nuclear factor-kappa B p65 (NF-κB p65) protein expressions. RESULTS: Seven major compounds of PDL were analyzed simultaneously. The treatment of PDL could attenuate LPS-induced histopathological damage of tracheal tissues, followed by reducing pro-inflammation mediators including TNF-α and IL-6 in serum and supernatant of tracheal tissue. LPS-induced nitroxidative stress including NO content and iNOS expression was inhibited significantly by PDL. Furthermore, PDL also down-regulated NF-kB p65 phosphorylation and TLR4 expressions. CONCLUSION: The results indicated that the PDL had a protective effect on LPS-induced respiratory inflammation injury in mice. Our findings for the first time provide experimental evidence for the application of PDL on respiratory inflammation injury in clinical practice.

Components of Goutengsan in Rat Plasma by Microdialysis Sampling and Its Protection on Aß1-42-Induced PC12 Cells Injury.

Goutengsan, a Chinese herbal formula, potential protection on Alzheimer's disease (AD) has been less reported. In current study, we investigated the protection of Goutengsan on Aß1-42-induced pheochromocytoma-derived cells (PC12). Furthermore, the components from Goutengsan in rat plasma were identified by microdialysis (MD) for in vivo sampling. Meanwhile, the protection of components identified was also verified. At last, we found that Goutengsan has a potential protective effect on Aß1-42-induced PC12 cells via reducing cells damage and increasing cells vitality as well as six components (pachymic acid, liquiritin, rhynchophylline, isorhynchophylline, corynoxeine, and isocorynoxeine) which may be effective components. This study helps to understand the treatment of Goutengsan for AD and would facilitate the clinical and further studies for this formula.

Protection of tauroursodeoxycholic acid on high glucose-induced human retinal microvascular endothelial cells dysfunction and streptozotocin-induced diabetic retinopathy rats.

ETHNOPHARMACOLOGICAL RELEVANCE: Tauroursodeoxycholic acid (TUDCA), one of the main ingredients from bear gall which hold "Clearing heat and detoxification, Removing liver fire for improving eyesight" functions, is formed by the conjugation of ursodeoxycholic acid (UDCA) with taurine. However, the limited information of TUDCA on protecting diabetic retinopathy (DR) has been known. The present study was conducted to evaluate the protection of TUDCA on high glucose-induced human retinal microvascular endothelial cells (HRMECs) dysfunction and streptozotocin (STZ)-induced diabetic retinopathy (DR) rats and the possible mechanism underlying was also explored. MATERIALS AND METHODS: The proliferation of high glucose-induced HRMECs was determined by MTT assay. DR rats' model was established by an administration of high-glucose-fat diet and an intraperitoneal injection of STZ (30mg/kg). The cell supernatant and rats' serum were collected for the assays of NO content by ELISA kits. Retinas were stained with hematoxylin and eosin (HE) to observe pathological changes. Immunohistochemical assay was applied to examine the protein expression of ICAM-1, NOS, NF-κB p65 and VEGF in rat retinas. Furthermore, western blot analysis was carried out to examine the protein expression of ICAM-1, NOS, NF-κB p65 and VEGF in high glucose-induced HRMECs. RESULTS: After treating with TUDCA, high glucose-induced HRMECs proliferation could be significantly inhibited. TUDCA (5.0µM, 25.0µM and 125.0µM) could decrease NO content in high glucose-induced HRMECs. Furthermore, TUDCA (500mg/kg/d and 250mg/kg/d) also decrease NO content in serum of DR rats. Additionally, both immunocytochemistry analysis and western blot analysis showed that the over-expression of ICAM-1, NOS, NF-κB p65 and VEGF were significantly decreased by TUDCA. CONCLUSION: The data indicated that TUDCA could ameliorate DR by decreasing NO content and down-regulating the protein expression of ICAM-1, NOS, NF-κB p65 and VEGF. Thus, our experimental results suggested that TUDCA might be a potential drug for the prevention and treatment of DR.

[Effect of Epimedii Herba alcohol extract on inhibition of lung tumor growth and immunomodulatory].

To observe the effect of Epimedii Herba alcohol extract (HE) on tumor growth of lung cancer by establishing the model of Lewis tumor-bearing mice, ELISA method was used to detect the levels of TNF-α, IL-10, IL-17, IL-2 in serum. Ki67 and P53 protein expression was detected in lung cancer tissues by using Western blot assay method and immunohistochemical assay method. The experimental results showed that HE has certain inhibitory effect on Lewis lung cancer tumor growth, and it can reduce the levels of TNF-α, IL-10 and IL-17 in serum, improve the level of IL-2,significantly decrease the expression of Ki67, and significantly increase P53 expression. HE has obvious inhibitory effect against lung cancer, and has the ability to improve immune regulating effect. This study reveals the anti-lung cancer effect of HE may be related to its ability of improving immunity, thus provides the basis for further research on anti-lung cancer effect of HE.

[Protective effect of seven kinds of phenolic acids of total salvianolic acids components on human umbilical vein endothelial cells with hypoxic injury].

In this paper, human umbilical vein endothelial cells (HUVEC) hypoxic injury models were established by sodium dithionite (Na2S2O4). With the protective effects of total salvianolic acids components (TSAC) against oxidative damage of HUVEC as a starting point, cellviability was measured by MTT colorimetric method； intracellular superoxide dismutase (SOD) activity, malondialdehyde (MDA), lactatedehydrogenase (LDH) level, nitric oxide (NO) level, interleukin6 (IL-6), and human tumor necrosis factor α (TNF-α) expression were measured by kits, to investigate the effect of seven kinds of phenolic acids of TSAC on Na2S2O4-induced HUVEC hypoxic injury. Based on the "component structure" theory, the contribution of the main components in TSAC for the protective effect on hypoxic injury of HUVEC was studied. The results showed that salvianolic acid B, posrnarinic acid A, salvianolic acid A, lactic acid, and lithospermic acid in TSAC play a larger role in protective effect of hypoxic injury of HUVEC. These five components were administered in combinations respectively, and it was concluded that the four components including salvianolic acid B, posrnarinic acid A, salvianolic acid A, lactic acid could be used as the representative components of TSAC.

Screening for anti-inflammatory components from Corydalis bungeana Turcz. based on macrophage binding combined with HPLC.

BACKGROUND: Corydalis bungeana Turcz. (CB; family: Corydalis DC.) is an anti-inflammatory medicinal herb used widely in traditional Chinese medicine (TCM) for upper respiratory tract infection, etc., but its anti-inflammatory active molecules are unknown. This study was designed to screen for the anti-inflammatory components from CB based on macrophage binding combined with HPLC. METHODS: Xylene-induced ear edema in mouse and carrageenan-induced hind-paw edema in rats were used to evaluate the anti-inflammatory activity of CB. The macrophage binding with high-performance liquid chromatography (HPLC) analysis and HPLC-MS were established to screen the potential active compounds. ELISA kits were performed to measure the levels of IL-6, IL-10, TNF-α and NO in RAW 264.7 macrophages culture media. RESULTS: The alkaloid extract of CB could inhibit significantly xylene-induced ear edema in mouse and carrageenan-induced hind-paw edema in rats. Two components binded to RAW 264.7 cell were identified as 12-hydroxycorynoline and corynoline. Bioassays demonstrated that these two compounds significantly inhibited LPS-induced IL-6, IL-10, TNF-α and NO levels. CONCLUSIONS: The results suggest that corynoline and 12-hydroxycorynoline contribute to the anti-inflammatory effects of the alkaloid extract of CB. Our findings suggest that these two compounds can be used as candidate for anti-inflammatory drugs.

[Simultaneous determination of seven components in Pudilan Xiaoyan oral liquid by HPLC].

OBJECTIVE: To establish an HPLC method for the content determination of baicalin, wogonin, chlorogenic acid, caffeic acid, cichoric acid, corynoline and adenosine in Pudilan Xiaoyan oral liquid. METHOD: The analysis was performed on a Phenomenex Luna C18 column (4.6 mm x 250 mm, 5 µm) with a gradient mobile phase of methanol-0.1% trifluoroacetic acid solution system at flow rate of 1.0 mL · min(-1). The detective wavelength was at 280 nm. The column temperature was 30 °C. RESULT: The standard curves of seven studied components show good linearity in their concentration ranges with r ≥ 0.999 6. The average recovery was 98.73%-102.1% with RSD less than 2.6%. CONCLUSION: The method is rapid, simple and accurate, and can be applied for the quality control of Pudilan Xiaoyan oral liquid.