OBJECTIVE To establish a method for determining the content of multiple components in Astragalus membranaceus,compare the content differences of Astragalus membranaceus in different regions and with different extraction methods in Gansu,and e-valuate the quality of Astragalus membranaceus from different origins of Gansu.METHODS Astragalus membranaceus samples from 40 origins in Gansu were collected on site.The content of Astragaloside Ⅳ,Calycosin-7-O-β-D-glucopyranoside,Ononin,Form-ononetin and Calycosin was determined by HPLC external standard method,and the quality differences of Astragalus membranaceus from 40 origins of Gansu were analyzed by combining bar graphs and line graphs.RESULTS Weiyuan County and Min County of Dingxi City showed high Astragaloside Ⅳ content in both extraction methods,indicating that the Astragalus membranaceus in these two regions has advantages in Astragaloside Ⅳ as an active component;for Calycosin-7-O-β-D-glucopyranoside and Ononin,although the performance of different regions in different extraction methods was different,in general,Weiyuan County and Min County of Dingxi City and Longnan City showed high content in both extraction methods.Water extraction method was more conducive to the extraction of Calycosin-7-O-β-D-glucopyranoside and Ononin,while alcohol extraction method was more conducive to the extraction of Astragalo-side Ⅳ,Calycosin and Ononin.CONCLUSION The content of chemical components of Astragalus membranaceus from different ori-gins is quite different,and the choice of extraction method has a significant effect on the content of active components,while the change trend of different compounds in the two extraction methods is basically the same.The established HPLC multi-component content de-termination method of Astragalus membranaceus is stable and reliable,which can provide a scientific basis for the quality control and comprehensive evaluation of Astragalus membranaceus medicinal materials.

Objective To investigate the effect of wogonin(WOG)on retinal ganglion cell(RGC)damage in chron-ic glaucoma rats and its relationship with the regulation of the JAK2/STAT3 signaling pathway.Methods A chronic glau-coma rat model was established in SPF-grade healthy male SD rats(50 modeled successfully).Successfully modeled rats were randomly divided into 6 groups(n=10 per group).The model group received an equal volume of normal saline(once daily,10 mL·kg-1 by gavage and 1 mL·kg-1 by intraperitoneal injection).The low-dose WOG group(L-WOG),medium-dose WOG group(M-WOG),and high-dose WOG group(H-WOG)received WOG at 7 mg·kg-1,14 mg·kg-1,and 28 mg·kg-1 by gavage,respectively(once daily,gavage volume 10 mL·kg-1).The high-dose WOG+Colivelin group(H-WOG+Colivelin)received WOG by gavage(28 mg·kg-1,once daily,10 mL·kg-1)plus intraperitoneal injection of the JAK2/STAT3 pathway activator Colivelin(1 mg·kg-1,once daily,injection volume 1 mL·kg-1).An additional 10 normal rats served as the sham-operated control group(Control group),undergoing surgery to expose the superior episcleral vein without cauterization,and receiving normal saline postoperatively in the same manner as the model group.All treatments lasted for 8 weeks.Intraocular pressure was measured using a tonometer.Retinal morphological changes and RGC apopto-sis were assessed by hematoxylin-eosin(HE)staining and TUNEL staining,respectively.Superoxide dismutase(SOD)ac-tivity and malondialdehyde(MDA)content in retinal tissue were measured by ELISA.Protein expression changes in the JAK2/STAT3 signaling pathway in retinal tissue were detected by Western blot.Results Retinal structure was normal in the Control group,with clear layers and densely arranged cells.Compared to the Control group,the model group showed significantly thinner retinal tissue,disorganized structure,sparsely arranged cells,and vacuolation.Compared to the model group,the L-WOG,M-WOG,and H-WOG groups exhibited increased retinal thickness and more orderly cell arrangement.Compared to the H-WOG group,the H-WOG+Colivelin group showed relatively thinner retinal tissue and sparser cell ar-rangement.After intervention,the RGC apoptosis rate in the model group(7.32％±1.07％)was significantly higher than that in the Control group(0.61％±0.12％)(P＜0.05).The apoptosis rates in the L-WOG group(5.67％±0.63％),M-WOG group(3.34％±0.75％),and H-WOG group(1.12％±0.35％)were significantly lower than that in the model group(all P＜0.05),showing a dose-dependent decreasing trend.The apoptosis rate in the H-WOG+Colivelin group(6.83％±1.56％)was significantly higher than that in the H-WOG group(P＜0.05).Compared to the Control group,SOD activity in the retinal tissue of the model group was significantly decreased(P＜0.05),while MDA content was significantly increased(P＜0.05).Compared to the model group,SOD activity was significantly increased and MDA content was significantly de-creased in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05),with this effect being dose-dependent(i.e.,H-WOG group＞M-WOG group＞L-WOG group).Compared to the H-WOG group,the H-WOG+Colivelin group showed signifi-cantly decreased SOD activity and significantly increased MDA content(P＜0.05).Compared to the Control group,the ra-tios of p-JAK2/JAK2 and p-STAT3/STAT3 expression in the retinal tissue of the model group were significantly upregulated(all P＜0.05).Compared to the model group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly downregulated in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05).Compared to the H-WOG group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly upregulated in the H-WOG+Colivelin group(all P＜0.05).Conclusion WOG may alleviate retinal tissue damage in chronic glaucoma rats by inhibiting the JAK2/STAT3 signaling pathway,thereby reducing oxidative stress and RGC apoptosis.

Objective To investigate the effect of wogonin(WOG)on retinal ganglion cell(RGC)damage in chron-ic glaucoma rats and its relationship with the regulation of the JAK2/STAT3 signaling pathway.Methods A chronic glau-coma rat model was established in SPF-grade healthy male SD rats(50 modeled successfully).Successfully modeled rats were randomly divided into 6 groups(n=10 per group).The model group received an equal volume of normal saline(once daily,10 mL·kg-1 by gavage and 1 mL·kg-1 by intraperitoneal injection).The low-dose WOG group(L-WOG),medium-dose WOG group(M-WOG),and high-dose WOG group(H-WOG)received WOG at 7 mg·kg-1,14 mg·kg-1,and 28 mg·kg-1 by gavage,respectively(once daily,gavage volume 10 mL·kg-1).The high-dose WOG+Colivelin group(H-WOG+Colivelin)received WOG by gavage(28 mg·kg-1,once daily,10 mL·kg-1)plus intraperitoneal injection of the JAK2/STAT3 pathway activator Colivelin(1 mg·kg-1,once daily,injection volume 1 mL·kg-1).An additional 10 normal rats served as the sham-operated control group(Control group),undergoing surgery to expose the superior episcleral vein without cauterization,and receiving normal saline postoperatively in the same manner as the model group.All treatments lasted for 8 weeks.Intraocular pressure was measured using a tonometer.Retinal morphological changes and RGC apopto-sis were assessed by hematoxylin-eosin(HE)staining and TUNEL staining,respectively.Superoxide dismutase(SOD)ac-tivity and malondialdehyde(MDA)content in retinal tissue were measured by ELISA.Protein expression changes in the JAK2/STAT3 signaling pathway in retinal tissue were detected by Western blot.Results Retinal structure was normal in the Control group,with clear layers and densely arranged cells.Compared to the Control group,the model group showed significantly thinner retinal tissue,disorganized structure,sparsely arranged cells,and vacuolation.Compared to the model group,the L-WOG,M-WOG,and H-WOG groups exhibited increased retinal thickness and more orderly cell arrangement.Compared to the H-WOG group,the H-WOG+Colivelin group showed relatively thinner retinal tissue and sparser cell ar-rangement.After intervention,the RGC apoptosis rate in the model group(7.32％±1.07％)was significantly higher than that in the Control group(0.61％±0.12％)(P＜0.05).The apoptosis rates in the L-WOG group(5.67％±0.63％),M-WOG group(3.34％±0.75％),and H-WOG group(1.12％±0.35％)were significantly lower than that in the model group(all P＜0.05),showing a dose-dependent decreasing trend.The apoptosis rate in the H-WOG+Colivelin group(6.83％±1.56％)was significantly higher than that in the H-WOG group(P＜0.05).Compared to the Control group,SOD activity in the retinal tissue of the model group was significantly decreased(P＜0.05),while MDA content was significantly increased(P＜0.05).Compared to the model group,SOD activity was significantly increased and MDA content was significantly de-creased in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05),with this effect being dose-dependent(i.e.,H-WOG group＞M-WOG group＞L-WOG group).Compared to the H-WOG group,the H-WOG+Colivelin group showed signifi-cantly decreased SOD activity and significantly increased MDA content(P＜0.05).Compared to the Control group,the ra-tios of p-JAK2/JAK2 and p-STAT3/STAT3 expression in the retinal tissue of the model group were significantly upregulated(all P＜0.05).Compared to the model group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly downregulated in the L-WOG,M-WOG,and H-WOG groups(all P＜0.05).Compared to the H-WOG group,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 expression were significantly upregulated in the H-WOG+Colivelin group(all P＜0.05).Conclusion WOG may alleviate retinal tissue damage in chronic glaucoma rats by inhibiting the JAK2/STAT3 signaling pathway,thereby reducing oxidative stress and RGC apoptosis.

OBJECTIVE To establish a method for determining the content of multiple components in Astragalus membranaceus,compare the content differences of Astragalus membranaceus in different regions and with different extraction methods in Gansu,and e-valuate the quality of Astragalus membranaceus from different origins of Gansu.METHODS Astragalus membranaceus samples from 40 origins in Gansu were collected on site.The content of Astragaloside Ⅳ,Calycosin-7-O-β-D-glucopyranoside,Ononin,Form-ononetin and Calycosin was determined by HPLC external standard method,and the quality differences of Astragalus membranaceus from 40 origins of Gansu were analyzed by combining bar graphs and line graphs.RESULTS Weiyuan County and Min County of Dingxi City showed high Astragaloside Ⅳ content in both extraction methods,indicating that the Astragalus membranaceus in these two regions has advantages in Astragaloside Ⅳ as an active component;for Calycosin-7-O-β-D-glucopyranoside and Ononin,although the performance of different regions in different extraction methods was different,in general,Weiyuan County and Min County of Dingxi City and Longnan City showed high content in both extraction methods.Water extraction method was more conducive to the extraction of Calycosin-7-O-β-D-glucopyranoside and Ononin,while alcohol extraction method was more conducive to the extraction of Astragalo-side Ⅳ,Calycosin and Ononin.CONCLUSION The content of chemical components of Astragalus membranaceus from different ori-gins is quite different,and the choice of extraction method has a significant effect on the content of active components,while the change trend of different compounds in the two extraction methods is basically the same.The established HPLC multi-component content de-termination method of Astragalus membranaceus is stable and reliable,which can provide a scientific basis for the quality control and comprehensive evaluation of Astragalus membranaceus medicinal materials.

Objective To evaluate the effects of exendin-4 on glial brillary acidic protein (GFAP ) and interleukin-1β(IL-1β) expression in hippocampi of aged rats .Methods Forty-eight healthy male Sprague-Dawley rats ,aged 22-24 weeks ,weighing 500-700 g ,were randomly divided into 4 groups (n=12 each) using a random number table:control group (group C ) ,exendin-4 group (group E ) ,operation group (group O ) and exendin-4 plus operation group (group OE) .The rats were anesthetized with intraperitoneal fentanyl and droperidol .Groups C and E did not receive anesthesia or splenectomy .In O and OE groups ,splenectomy was carried out .In E and OE groups , exendin-4 5 μg/kg was injected intraperitoneally at 30 min before skin incision and 12 h after operation .C and O groups received the equal volume of normal saline instead of exendin-4 .Learning and memory function was assessed using Morris water maze test (escape latency (EL) and total swimming distance (TSD) at 1 day before operation (T0 ) .The fasting blood glucose was measured after anesthesia (T1 ) ,at the end of operation (T2 ) and on postoperative day 1 (T3 ) .The rats were sacrificed after assessment of the cognitive function at T 3 and the hippocampi were removed for determination of the expression of GFAP (by immuno-histochemistry ) and IL-1β(by Western blot ) .Results There was no significant difference in the EL and TSD at T0 between the four groups ( P>0.05) .Compared with group C ,the EL and TSD were significantly prolonged at T3 and fasting blood glucose was increased at T2 ,3 ,and the expression of IL-1βand GFAP was up-regulated at T3 in O and OE groups ( P<0.05) .Compared with group O ,the EL and TSD were significantly prolonged at T3 and fasting blood glucose was decreased at T2 ,3 ,and the expression of IL-1βand GFAP was down-regulated at T3 in group OE ( P<0.05) . Conclusion Exendin-4 can improve the postoperative cognitive function of aged rats by inhibiting inflammatory responses in hippocampi and maintaining stable perioperative blood glucose .

Aim To investigate the anti-apoptotic effect of NGF on H9 c2 cardiac myocytes in a hypoxia / reox-ygenation injury model and its mechanism. Methods The H9 c2 cardiac myocytes were randomly divided into five groups:control group ( C group) , hypoxia/reoxy-genation group ( H/R group) , NGF group ( N group) , NGF+LY294002 group ( N+L group) and LY294002 group( L group) . Each group received the correspond-ing treatment. Cell survival rate was tested by cell counter kit-8 methods. Apoptotic rate was evaluated by propidium iodide ( PI ) staining and flow cytometry (FCM). The levels of Caspase-12, p-Akt/Akt were e-valuated by Western blot. Results The NGF group could significantly protect the H9 c2 cardiac myocytes under the hypoxia / reoxygenation injury with increased cell survival rate. It also decreased the apoptotic per-centage, upregulated the level of p-Akt/Akt and inhib-ited the expression of Caspase-12 . As the specific in-hibitor of PI3k receptor, LY294002 decreased the level of p-Akt. Conclusion NGF has the effect of anti-ap-optosis on H9 c2 cardiac myocytes exposed to hypoxia /reoxygenation injury via PI3k-Akt signal pathway.