Effect And Mechanism Of Shuangshi Tonglin Capsule On Prostatic Hyperplasia In Rats

ObjectiveBy establishing castration combined with subcutaneous injection of testosterone propionate(TP),the rat model of prostatic hyperplasia(BPH)was established,and the efficacy and mechanism of Shuangshi Tonglin capsule on BPH rats were explored,providing theoretical basis for Shuangshi Tonglin capsule to treat prostatic hyperplasia.MethodsSixty-six SD rats were randomly divided into 6 groups.Except for sham operation group,BPH rat models were induced by castration combined with subcutaneous injection of TP,and were randomly divided into model group,Shuangshi Tonglin capsule low-dose,medium-dose and high-dose groups(0.625 g/kg,1.25 g/kg,2.5 g/kg)and finastomide group.At the same time,drug intervention was carried out for 28 consecutive days,and indicators were detected.(1)To explore the effect of Shuangshitonglin Capsule on BPH rats.The wet weight and prostate index were determined.The pathological changes of prostate tissue were observed by HE staining and Masson staining.Sex hormone indexes were detected by ELISA kit.(2)To explore the regulatory effect of Shuangshi Tonglin Capsules on ROS/NLRP3 signaling pathway in BPH rats.The levels of TNF-α and IL-1β in serum and prostate tissue were determined by ELISA kit.The levels of MDA,SOD,GSH-Px and CAT in serum and prostate tissue of rats were detected by the kit.ROS levels in prostate tissues were observed by immunofluorescence.The expression of NLRP3 inflammasome in prostate tissue was detected by immunohistochemistry.Western blotting was used to detect the expression of NOX4,NOX2,NLRP3 inflammasome and related proteins in prostate tissue of rats.(3)To explore the regulatory effect of Shuangshi Tonglin Capsule on TGF-β1/Smad signaling pathway in BPH rats.The expressions of TGF-β1/Smad related proteins and genes in prostate tissues were detected by Western blotting and RT-PCR.The protein and gene expressions of α-SMA and Collagen I in prostate tissues were detected by Western blotting and RT-PCR.The expression of E-cadherin and Vimentin in prostate tissues were detected by immunohistochemistry.Results(1)Shuangshi Tonglin capsule can improve the increase of prostate index in BPH rats induced by TP,improve the pathological injury of prostate tissue and reduce collagen deposition,and reduce the levels of T and DHT in serum and prostate tissue of rats.(2)Shuangshi Tonglin capsule can reduce the levels of inflammatory factors TNF-α and IL-1β,increase the activities of SOD,CAT and GSH-Px,and decrease the activities of MDA in BPH rats.Immunofluorescence results showed that the ROS expression decreased after the intervention of Shuangshi Tonglin capsule.Western blotting results showed that Shuangshi Tonglin capsule reduced the expression of NOX4 and NOX2 protein in prostate tissue.The above results suggest that Shuangshi Tonglin capsule may reduce the peroxide damage of the body by down-regulating the expression of NOX2 and NOX4.Western blotting results showed that Shuangshi Tonglin capsule could decrease the expression of NLRP3 inflammasome,ASC,Cleaved Caspase-1,and IL-1β,suggesting that Shuangshi Tonglin capsule could inhibit the expression of NLRP3 inflammasome and related proteins.(3)Western blotting results showed that Shuangshi Tonglin capsule could downregulate the expression of TGF-β1 and p-Smad2/3.RT-PCR showed the same trend,suggesting that Shuangshi Tonglin capsule could inhibit TGF-β1/Smad pathway.Western blotting results showed that Shuangshi Tonglin capsule could down-regulate the expressions of α-SMA and Collagen I.RT-PCR showed the same trend.Immunohistochemical results showed that Shuangshi Tonglin capsule can down-regulate the epithelial-interstitium transition(EMT)related protein Vimentin and up-regulate the expression of E-cadherin,suggesting that Shuangshi Tonglin capsule can inhibit the deposition of collagen in prostate tissue and block the process of EMT.Conclusion(1)Shuangshi Tonglin capsule reduced prostate index,reduced collagen deposition,improved pathological damage of prostate tissue,and reduced T and DHT levels in serum and prostate tissue of BPH rats.(2)The mechanism of the improvement of BPH may be related to inhibiting the activation of ROS/NLRP3 inflammassome pathway,reducing inflammatory factors and alleviating oxidative stress,inhibiting the activation of TGF-β1/Smad,downregulating the expression of Collagen I and α-SMA,and blocking the progression of EMT.