Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

BACKGROUND: Changing the local environment of spinal injury promotes the repair and regeneration of injured nerve and recovery of partial nervous function of spinal cord. Transplantation of olfactory ensheathing cells can improve the local internal environment of injured spinal cord.OBJECTIVE: To probe into the effect and safety of transplantation of olfactory ensheathing cells on functional repair of spinal cord and nerve in patients with obsolete spinal injury DESIGN: Self-control experiment.SETTING: Wards of the Department of Surgery, Taian Rongjun Hospital of Shandong Province.PARTICIPANTS: Totally 48 patients admitted for obsolete spinal injury in the Department of Surgery, Taian Rongjun Hospital, between June 2004 and July 2005 were recruited. There were 39 males and 9 females, aged 7 to 59 years with the mean of 36 years.METHODS: ①Cell culture: Olfactory bulb of aborted fetus was digested into single olfactory ensheathing cells, which were then cultured and puri fied for 1 to 2 weeks, and finally made into single cell suspension. ②Operation and cell transplantation: Under general anesthesia, the purified single cell suspension (about 0.05-0.20 mL) of olfactory ensheathing cells was injected into the corresponding spinal injury site through multiple points with home-made syringe of 0.45 mm in diameter. Stitches were taken out at postoperative 10 to 14 days. ③Evaluation of spinal function: Injury Scoring Standard made by American Spinal Injury Association (ASIA) was used for scoring, comparison and statistical analysis at postoperative 1 day and 2 weeks to 2 months. ④Spinal function of 48 patients was observed or followed up through telephone at postoperative 3 weeks to 1 year.MAIN OUTCOME MEASURES: Changes of postoperative sensory function of the patients. Changes of postoperative motor function of the patients. Changes of postoperative automatic nervous system of the patients.RESULTS: ①All the 48 patients had improvement in spinal function, and continued improved tendency was found in the observation and follow-up through telephone at postoperative 3 weeks to 1 year. ②Scoring by ASIA for sensory function was higher after operation than before operation (touch sensation: 56.9, 51.2, P ＜ 0.01; pain sensation: 55.2, 48.3, P ＜ 0.01). Sensory function was improved obviously at the lower shift of sensory level,generally more than 2 segments. ③Scoring by ASIA for motor function was higher after operation than before operation (44.8, 40.7, P ＜ 0.01), but the improvement was slow. ④Scoring by ASIA for automatic nervous system was higher after operation than before operation (18.0, 14.5, P ＜ 0.01); diaphoresis, increased enterokinesia and other automatic nervous system improved earliest.CONCLUSION: Transplantation of olfactory ensheathing cells promotes the spinal and neurofunctional recovery of patients with malignant spinal injury, and the therapeutic method is safe.