A puncture device sealing membrane (130) comprising forward and reverse grooves and an assembly (180). A sealing film (130) comprises a proximal opening (132), a distal opening (133), and a sealing wall extending from the distal opening (133) to the proximal opening (132). The distal opening (133) is formed by a sealing lip (134) and is used for receiving a device inserted therein and forming a gas-tight seal; the sealing wall comprises a proximal end surface and a distal end surface, and is a seamless sealing body formed by a plurality of forward grooves and a plurality of reverse grooves which surround the sealing lip (134) in an alternately forward and reverse manner. The forward grooves and reverse grooves feature functions such as increasing a circumferential length, reducing a wrapping area, improving lubrication reliability and increasing axial tensile rigidity, thereby greatly reducing frictional resistance and stick-slip.