ObjectiveTo evaluate the efficacy of Shexiang Baoxinwan in treating stable angina pectoris with Qi stagnation and blood stasis syndrome in patients with coronary artery disease （CAD） complicated with anxiety and depression and explore its underlying mechanisms. MethodsThis study employed a randomized， double-blind， and placebo-controlled clinical trial design. Patients admitted to the hospital were randomly assigned to the observation group and the control group， with 52 patients in each group. Patients in the observation and control groups received Shexiang Baoxinwan and placebo， respectively， both in combination with conventional Western medication. The dose was 45.0 mg， three times daily， for a total duration of eight weeks. The primary outcome was the Seattle Angina Questionnaire （SAQ） scores before and after treatment. Secondary outcomes included changes in traditional Chinese medicine （TCM） syndrome score， the patient health questionnaire-9 （PHQ-9）， generalized anxiety disorder-7 （GAD-7）， inflammatory markers ［interleukin-18 （IL-18）， interleukin-10 （IL-10）， tumor necrosis factor-alpha （TNF-α）， CD40， etc.］， monoamine neurotransmitters ［e.g.， dopamine （DA）］， vascular endothelial function markers ［e.g.， endothelin-1（ET-1）］， adipokines， and ischemia-modified albumin （IMA）. Adverse reactions were also recorded. ResultsA total of 92 patients completed the study， with 44 in the observation group and 48 in the control group. Compared with baseline， both groups showed significant decreases in PHQ-9， GAD-7， and TCM syndrome scores following treatment （P<0.05）， along with a significant increase in SAQ scores （P<0.05）. In the observation group， DA levels were significantly increased （P<0.05）， while levels of IL-18， TNF-α， CD40， ET-1， and IMA were decreased （P<0.05）. In contrast， the control group exhibited significantly increased CD40 levels （P<0.05）. Compared with the control group after treatment， the observation group showed significant improvements in the SAQ dimensions of physical limitation， angina stability， treatment satisfaction， and disease perception， as well as in TCM syndrome score， PHQ-9 score， IL-18， CD40， ET-1， and IMA （P<0.05）. No adverse reactions were observed in either group during treatment. ConclusionShexiang Baoxinwan can improve anxiety and depression， alleviate angina symptoms， and reduce TCM symptoms of Qi stagnation and blood stasis in CAD patients. The mechanism may involve anti-inflammation， improvement of vascular endothelial function， reduction of IMA， and increase of monoamine neurotransmitter levels.

ObjectiveTo evaluate the efficacy of Shexiang Baoxinwan in treating stable angina pectoris with Qi stagnation and blood stasis syndrome in patients with coronary artery disease （CAD） complicated with anxiety and depression and explore its underlying mechanisms. MethodsThis study employed a randomized， double-blind， and placebo-controlled clinical trial design. Patients admitted to the hospital were randomly assigned to the observation group and the control group， with 52 patients in each group. Patients in the observation and control groups received Shexiang Baoxinwan and placebo， respectively， both in combination with conventional Western medication. The dose was 45.0 mg， three times daily， for a total duration of eight weeks. The primary outcome was the Seattle Angina Questionnaire （SAQ） scores before and after treatment. Secondary outcomes included changes in traditional Chinese medicine （TCM） syndrome score， the patient health questionnaire-9 （PHQ-9）， generalized anxiety disorder-7 （GAD-7）， inflammatory markers ［interleukin-18 （IL-18）， interleukin-10 （IL-10）， tumor necrosis factor-alpha （TNF-α）， CD40， etc.］， monoamine neurotransmitters ［e.g.， dopamine （DA）］， vascular endothelial function markers ［e.g.， endothelin-1（ET-1）］， adipokines， and ischemia-modified albumin （IMA）. Adverse reactions were also recorded. ResultsA total of 92 patients completed the study， with 44 in the observation group and 48 in the control group. Compared with baseline， both groups showed significant decreases in PHQ-9， GAD-7， and TCM syndrome scores following treatment （P<0.05）， along with a significant increase in SAQ scores （P<0.05）. In the observation group， DA levels were significantly increased （P<0.05）， while levels of IL-18， TNF-α， CD40， ET-1， and IMA were decreased （P<0.05）. In contrast， the control group exhibited significantly increased CD40 levels （P<0.05）. Compared with the control group after treatment， the observation group showed significant improvements in the SAQ dimensions of physical limitation， angina stability， treatment satisfaction， and disease perception， as well as in TCM syndrome score， PHQ-9 score， IL-18， CD40， ET-1， and IMA （P<0.05）. No adverse reactions were observed in either group during treatment. ConclusionShexiang Baoxinwan can improve anxiety and depression， alleviate angina symptoms， and reduce TCM symptoms of Qi stagnation and blood stasis in CAD patients. The mechanism may involve anti-inflammation， improvement of vascular endothelial function， reduction of IMA， and increase of monoamine neurotransmitter levels.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.

Objective To investigate the effect and mechanism of Panax notoginseng saponins(PNS)in inhibiting c-Jun N-terminal protein kinase(JNK)/c-Jun signaling pathway activation to alleviate calcific aortic valve disease(CAVD)in mice.Methods Twenty-one male ApoE-/-mice aged 6 to 8 weeks were randomly divided into the model,PNS high-dose(60 mg/kg),and PNS low-dose(30 mg/kg)groups using the random number table method,with seven mice per group.Nine male C57BL/6 mice aged 6 to 8 weeks were used as the control group.Mice in the control group were fed a normal diet,whereas ApoE-/-mice were fed a high-fat diet for 12 weeks.After 12 weeks,three C57BL/6 and three ApoE-/-mice(one ApoE-/-mice from each group)were randomly selected to evaluate the CAVD modeling effect.After confirming successful modeling,the PNS high-and low-dose groups received daily intragastric PNS administration.The control and model groups were administered an equal volume of stroke-physiological saline solution by gavage for 4 consecutive weeks.The valve annulus diameter and peak velocity of the mice in each group were then detected using ultrasound.The degree of aortic valve calcification was evaluated using von Kossa and Alizarin Red S staining.The serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C)were detected by biochemical method.Inflammatory factor interleukin-4(IL-4),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),and interleukin-10(IL-10)levels were determined using an enzyme-linked immunosorbent assay.The expressions of calcification markers,runt-related transcription factor 2(RUNX2),and bone morphogenetic protein 2(BMP2)were detected using immunohistochemistry.Aortic valve cell apoptosis was evaluated using TUNEL staining,and JNK/c-Jun signaling pathway-related mRNA and mean fluorescence intensity were detected using quantitative real-time PCR and immunofluorescence,respectively.Results Compared with the control group,the mice in the model group showed an increase in serum TC,TG,LDL-C,TNF-α,and IL-1β levels,a decrease in IL-4 and IL-10 levels,a decrease in annulus diameter,an increase in peak flow velocity,and an increase in von Kossa and Alizarin Red S staining-positive areas.Additionally,the model group showed an increase in aortic valve cell apoptosis rate,an increase in BMP2 and RUNX2-positive rates,and an increase in JNK and c-Jun mRNA expression levels and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Compared to the model group,the PNS low-dose group showed a decrease in serum TC,LDL-C,and TNF-α levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in positive area in Alizarin Red S staining.Furthermore,the PNS low-dose group showed a decrease in BMP2 and RUNX2-positive rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).The PNS high-dose group showed an increase in HDL-C,IL-4 and IL-10 levels,a decrease in serum TC,LDL-C,TNF-α,and IL-1β levels,an increase in annulus diameter,a decrease in peak flow velocity,and a decrease in von Kossa and Alizarin Red S staining-positive areas and cell apoptosis rate.The PNS high-dose group also showed a decrease in BMP2 and RUNX2 positive staining rates,JNK and c-Jun mRNA expression levels,and p-JNK/JNK and p-c-Jun/c-Jun(P<0.05).Conclusion PNS may reduce valvular cell apoptosis,alleviate inflammation,and protect against aortic valve calcification in mice by inhibiting the activation of JNK/c-Jun signaling pathway.