升清降浊平衡调控RIP1/RIP3表达-促进肥胖急性胰腺炎大鼠腺泡细胞坏死-凋亡转换的机制

Severe acute pancreatitis (SAP) is a life-threatening disease with high mortality. As a chronic low-grade inflammation, obesity aggravates the inflammatory injury and mortality of SAP. Regulation of necrosis-apoptosis switch in the pancreatic acinar cell to ameliorate the inflammatory injury is key to lower the mortality of the disease. Regulation of RIP1/RIP3, a key molecular switch of necroptosis, helps promote necrosis-apoptosis switch in pancreatic acinar cell and alleviate pancreatic injury in AP. Sheng-jiang-san (SJS) could lower the body weight and relieve systematic inflammatory response in AP. Together with the preliminary study of herbal formula SJS with lowering the body weight of obese rats and protecting pancreatic inflammatory injury in obese AP rats and the hypothesis of formula tissue pharmacology, we hypothesized : SJS Sheng-Qing-Jiang-Zhuo could reverse necroptosis and promote necrosis-apoptosis switch in pancreatic acinar cell to alleviate inflammatory injuries via regulating RIP1/RIP3 expression in obese AP rats. We plan to set up obese AP model and select RIP1/RIP3 as therapeutic targets in pancreas, to firstly, compare the pharmacokinetic process and the concentrations of SJS in obese rats and obese AP rats. And secondly, illuminate the mechanism of SJS in regulating pancreatic acinar cell necrosis-apoptosis switch and inhibiting inflammatory response in obese AP rats. Then thirdly, elucidate the formula tissue pharmacology of SJS in protecting pancreatic injuries. And eventually, clarify the pharmacokinetic-pharmacodynamic mechanism of SJS in reversing necroptosis, promoting necrosis-apoptosis switch and relieving the inflammatory injuries in pancreatic acinar cells of obese AP rats.

重症急性胰腺炎（SAP）病死率高，肥胖加重炎症损伤并增加SAP病死率。调控胰腺腺泡细胞坏死-凋亡转换以减轻肥胖SAP炎症损伤是降低病死率的关键。调控程序性坏死关键分子开关RIP1/RIP3有助于促进SAP腺泡细胞坏死-凋亡转换以减轻炎症损伤；升降散降低体重并减轻SAP患者全身炎症反应；结合预实验升降散减轻肥胖大鼠体重和胰腺炎性损伤的基础和方剂组织药理学，假设：升降散升清降浊平衡RIP1/RIP3表达以逆转坏死并促进坏死-凋亡转换、减轻肥胖SAP炎症损伤。拟建立肥胖SAP模型并选择RIP1/RIP3细胞死亡分子开关：比较升降散在肥胖大鼠和肥胖SAP大鼠中药动学及胰腺组织内浓度；阐明升清降浊法促进胰腺腺泡细胞坏死-凋亡转换以抑制炎症反应的机理；明确升降散保护胰腺组织的方剂组织药理学。最终阐明升降散逆转胰腺腺泡坏死，促进坏死-凋亡转换以减轻炎症损伤的药动学-药效学机理。

重症急性胰腺炎（SAP）病死率高，肥胖加重炎症损伤并增加SAP病死率。调控胰腺腺泡细胞坏死-凋亡转换以减轻肥胖SAP炎症损伤可能是降低病死率的关键。而调控程序性坏死关键分子开关RIP1/RIP3表达可能有助于促进SAP腺泡细胞坏死-凋亡转换以减轻炎症损伤；结合临床升降散减轻SAP患者全身炎症反应及预实验升降散减轻肥胖大鼠体重和胰腺炎性损伤的基础，我们开展本实验以阐明升降散平衡调控RIP1/RIP3表达，逆转胰腺腺泡细胞坏死并促进坏死-凋亡转换以减轻肥胖SAP炎症损伤的机制。本实验通过建立肥胖SAP大鼠模型及细胞模型，比较升降散在肥胖大鼠和肥胖SAP大鼠中药动学及胰腺组织内浓度；阐明升降散促进胰腺腺泡细胞坏死-凋亡转换以抑制炎症反应的机理和靶点；并明确升降散保护胰腺组织的方剂组织药理学。研究结果发现：肥胖SAP大鼠胰腺RIP3活化明显增加，腺泡细胞坏死明显增多，炎症反应及胰腺病理损伤明显加重，而升降散能有效减少肥胖SAP大鼠胰腺腺泡细胞坏死，增加凋亡，减轻炎症反应及胰腺组织病理损伤，其作用可能与抑制程序性坏死关键分子RIP3的活化相关，这为进一步深入研究升降散减少肥胖SAP胰腺腺泡细胞坏死的确切分子机制奠定基础，并为临床采用升降散治疗肥胖SAP的有效性提供基础研究证据。另外，通过药动药效学分析，我们初步确定大黄酸可能是升降散减轻肥胖SAP炎症损伤的主要有效成分，这为进一步研究开发治疗肥胖SAP的有效药物提供方向。