1.Changes of A20 expression in mesangial cells of LPS-induced diabetic nephropathy rat model
Yaling LIANG ; Jiao CHEN ; Yang LONG ; Yanhui LI ; Qiuhan LI ; Fang FAN ; Yong XU
Tianjin Medical Journal 2016;44(4):444-448
Objective To observe the changes of A20 in mesangial cells of diabetic nephropathy (DN) rat model in?duced by lipopolysaccharide (LPS)-rat, and to explore its possible mechanism. Methods (1)Thirty health male Wistar rats were randomly divided into two group. Model rats were given streptozotocin (STZ) at a dose of 60 mg/kg by intraperitoneal in?jection. Rats in the control group received the same volume of citrate buffer in the same way. Levels of blood glucose and uri?nary microalbumin were detected in two groups at the 6th and the 8th week. Changes of renal pathology were observed by HE staining. Changes of protein A20 were observed by immunohistochemistry. (2) Expression changes of gene and proteins A20, nuclear factor (NF)-κB, IκB, IKKγand MCP-1 in renal cells treated with LPS were determined after treatment with different time points (0, 2, 4, 6, 12, 24, 48 and 72 h) and different concentrations (0.1, 1 and 10μg/L). Results (1) Levels of blood glucose and urinary microalbumin were significantly increased in model group compared with those of control group ( P <0.01). HE stainig showed that hyaline degeneration in tubular epithelial cells was found in model group, especially at the 8th week. Results of immunohistochemistry showed that expression of protein A20 significantly decreased in kidney tubules and nearly disappeared in glomerulus in model group compared with that of control group, which expressed less at the 8th week. (2) There was no significant difference in the expression of IKKγbetween different concentrations and different times. Com?pared with 0 h, the expression of A20 protein was increased at 2 h and 4 h, except that the expression of A20 protein in?creased after 6 h (P<0.05). Meanwhile NF-κB expression increased and IκB expression decreased in different time points (P<0.05). In addition, the expressions of A20 and IκB were decreased concentration-dependently (P<0.05). The expres?sion levels of NF-κB and MCP-1 were increased concentration-dependently (P<0.05). Conclusion A20 may involve in the development of diabetic nephropathy by regulating the NF-κB pathway.
2.Preparation and Characterization of Syringopicroside Solid Lipid Nanoparticles
Xiwu ZHANG ; Qiuhan LI ; Yingpeng LI ; Yongji LI ; Zuodi XU ; Jinjin DOU
China Pharmacy 2019;30(9):1168-1172
OBJECTIVE: To prepare Syringopicroside solid lipid nanoparticles (SYR-SLN), and optimize the formula and characterize SYR-SLN. METHODS: SYR-SLN were prepared by emulsion evaporation method. Using entrapment efficiency as index, based on single factor, orthogonal design was adopted to optimize the mass ratio of lecithin-monoglyceride, volume ratio of organ phase to water phase, poloxamer 188 (F68) concentration and drug dosage. The optimal formula technology was established to investigate entrapment efficiency, drug-loading amount, morphology, particle size, Zeta potential, stability, etc. RESULTS: The mass ratio of lecithin-monoglyceride was 3 ∶ 1; the volume ratio of organic phase to water phase was 1 ∶ 2; the concentration of F68 was 0.4%; drug dosage was 10 mg. The optimal formula included that monoglyceride 80 mg, lecithin 240 mg, 0.4% F68, syringopicroside 10 mg, absolute ethyl alcohol 5 mL, distilled water 10 mL, emulsification temperature at 65℃ and stirring at 600 r/min. Encapsulation efficiency of SYR-SLN was (42.35±0.60)% (n=3); drug-loading amount was (5.33±0.03)% (n=3); SYR-SLN had a spherical morphology and was evenly distributed. The average particle size was (180.30±5.31) nm with Zeta potential of (-41.9±0.8) mV, and the SYR-SLN could maintain stable for 15 days at 4℃. CONCLUSIONS: SYR-SLN is prepared successfully, and the technology is simple with high encapsulation efficiency.